· 1 CONTENT Page Foreword 3 General Information 4 Welcome Address 6 Assoc. Prof. Dr. Siree...

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CONTENT

Page

Foreword 3

General Information 4

Welcome Address 6

Assoc. Prof. Dr. Siree Chaiseri

Interim Vice President, KU, Thailand

Opening Message 8

Dr. Takashi Nagai

Deputy Director, FFTC

Tentative Schedule 13

Speaker Biodata

Paper Presentation

Keynote Speaker

1. The techno rice: an industrial application of supercritical 20

fluid technology in Taiwan

Dr.Yi-Jen Liaw

Session I - Overview: Risk assessment and risk management of

controlling and limiting agrochemical

2. Analysis and risk assessment of pesticides in Korean 34

agricultural products Dr.Kwang-Geun Lee

3. Current situation of pesticides use in Indonesian 49

agricultural products

Ms. Anna Rahmianna

4. Current use of pesticides in the agricultural products 61

of Cambodia

Dr.Preap Visarto

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5. Pesticide residues in food and the environment in the Philippines: 71

risk assessment and management

Ms. Cristina Bajet

Session II - Novel technologies in practical: Agrochemicals residues

Reduction and food safety improvement

6. Application of ultrasound technology for agricultural product 84

improvement and environmental renovation

Dr. Nomura Nakao

7. Using sanitizer and fine bubble technologies to enhance food safety 90

Dr. Warapa Manakarnchanakul

Session III - Analytical approaches for agrochemical determinations

8. Application of non-target analysis by high-resolution 111

mass spectrometry

Dr. Chung-Huang Wang

9. Chemometric approach to the optimization of HS-SPME/GC-MS 125

for the determination of multiclass pesticide residues in fruits

and vegetables

Dr.Tan Guan Huat

Session IV –Key success for Good Agricultural Practices (GAP)

10. Thai good agricultural practice (GAP) 147

Dr. Rungnapa Korpraditskul

11. Development of good agricultural practice (GAPs) model for tea, 157

rice and vegetable in Vietnam

Dr. Dao Bach Khao

12. Producers’ perceptions of public Good Agricultural Practices and 172

their pesticide use: the case of Durian farming in Pahang, Malaysia

Dr. Yuichiro Amekawa

Organizing Committee 192

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FOREWORD

Warmth greetings from Kasetsart University, Thailand

As now it is a time of 2015 FFTC-KU International Workshop on Risk Management on

Agrochemicals through Novel Technologies for Food Safety in Asia. I would like to

welcome all of you to this important workshop in Sampran Riverside, Nakorn Pathom.

Agrochemicals or pesticides play a vital role on the efficient and economic production

of agricultural products. Pesticides are known to be widely used in most agricultural area

to control pests. Although these chemicals are safe when properly used under a limited

level, chemical residues are found to affect consumers’ health. Therefore, a pest control

measure either by using pesticide or alternative measure should be implemented to

safeguard both consumers’ and farmers’ health. To become less dependent on the

chemical use, integrated pest management (IPM) and organic farming can be

alternatively used.

I am very happy and proud indeed to see the gathering of scientists, researchers and

private sections to share and exchange findings on the subject matter in this coming

workshop. I believe this meeting will be invaluable and be able to identify necessary

research agenda as well as policy options to be taken home for implementation.

I and all organizing committee would like to express our sincere gratitude to Interim Vice

President Assoc. Prof. Dr. Siree Chaiseri for honoring us to preside over the opening

ceremony of this workshop. Our special thanks also go to all guest speakers, participants

to participate in this event. We wish all of you to have a productive conference and enjoy

the time with us.

Assistant Professor Dr. Warapa Mahakarnchanakul

Kasetsart University

Co-Organizer of the FFTC-KU 2015 International Workshop

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GENERAL INFORMATION OF SEMINAR

Introduction

Agrochemicals play an important role on the efficient and economic production of

agricultural products. Major group of agrochemicals are pesticides including herbicides,

insecticides and fungicides. Pesticides are part of agrochemicals widely used in most

agricultural areas. The chemicals are used to control pests such as insects, rodents,

microorganisms and weeds. These chemicals are considered safe when used under a

limitation level with proper application procedure. Nonetheless many of the produce in

markets are still found to contain chemical residues in which lead to more awareness on

the effects of residues on consumer health. Some chronic diseases such as cancer,

diabetes, Parkinsons, and Alzheimers are reported to have direct or indirect relationships

with pesticide exposures and development of diseases. Climate change also known to be

a factor that influences an increasing number of pests, which lead to crop damages.

Therefore, a pest control measure either by using pesticides or other alternative measures

should be done. Awareness of the health threats from the increasing use of pesticides and

other farm chemicals are on the rise in the Asian and the Pacific region. This include the

consumers’ and the farmers’ health. Integrated pest management (IPM) and organic

farming are alternative choices to move the agrochemical users to avoid or minimize the

chemicals used in their production. Moreover, cost of production and competitiveness

enhancement with other regular farm produce are also critical. Novel technologies should

be more broadly applied to reduce the residues in fresh produce e.g. ozonation,

microbubbles, hydrostatic pressure, etc.

Faculty of Agro-Industry and Faculty of Agriculture at Kasetsart University and

Food and Fertilizer Technology Center for the Asian and Pacific Region with their

network are collaborating to develop an alternative measure for proper pesticide

management for farm produces aiming at safe food production, nonetheless, the market

share enhancement for these chemical-free produces will also be discussed and shared

with the network.

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Objective

1. To bring together community leaders, scientists, and policy leaders to promote and

enhance programmatic collaborations to more effectively address regional, national and

local responses to minimize chemical utilization on crop production and increase organic

farming.

2. To create an active network for assessment and monitoring chemical utilization and

organic farming in Asia especially in the country that plays important role as a global

food supplier. Such information will help the risk-based strategic plan for food safety

management. The network will be created and exchanged for information among

members using database and website as a key decision tool and organized by an elected

network cooperator.

Speakers

1. International experts from Cambodia, Indonesia, Japan, Korea, Malaysia, Philippines,

Vietnam, Taiwan, Thailand and representatives from FFTC.

2. Local experts from the Center of Excellence Kasetsart Logistics and food logistics

industry

Organizers

The Agrochemical international workshop FFTC-KU 2015 is jointly organized by The

Food and Fertilizer Technology Center for the Asian and Pacific Region (FFTC) and

Kasetsart University (KU), Thailand. Leading international speakers from Cambodia,

Indonesia, Japan, Korea, Malaysia, Philippines, Vietnam, Taiwan, Thailand and

representatives from FFTC as well as participants in agrochemical research area

participated in this event

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WELCOME ADDRESS

Associate Professor Dr. Siree Chaiseri

Interim Vice President of Kasetsart University (KU), Thailand

On behalf of Kasetsart University, may I take this opportunity to extend our warm

welcome to all honourable speakers and distinguished participants to the FFTC-KU

International workshop on “Risk Management on Agrochemicals through Novel

Technologies for Food Safety in Asia” which is a joint effort of cooperation between

Kasetsart University and Food and Fertilizer Technology Center for the Asian and Pacific

Region or FFTC, Taiwan.

May I also express our sincere thanks and appreciation to FFTC, Taiwan who entrusts

and honors Kasetsart University to be the host of this important workshop.

Kasetsart University, as a national research university specializing in many areas

including agricultural science, agro-industry, engineering, environment, and food safety,

has a major goal become a leading research university in Asia. The University, therefore,

has been investing her budget to strengthen her research capacities and to strengthen

research collaboration with her international partners for the academic excellence and the

sustainable development.

Food safety is one of the world’s major concerns the need urgent cooperation to mitigate

the risks involved. Currently, Thailand and many Asian countries play an important role

as the world’s major food producers; therefore, the assessment of food contamination and

other related issues have become more important in the region. Agrochemicals are

considering safe when used under a limited level with proper application procedure.

Nonetheless, many produces in markets are still found with chemical residues leading to

awareness of effects on consumer health. Some chronic diseases such as cancer, diabetes,

Parkinson, and Alzheimer are reported on the relationship between pesticide exposures

and the diseases development.

It is a good opportunity that this workshop is held to be a forum for the exchange of

knowledge and experiences of research studies regarding “Risk Management on

Agrochemicals through Novel Technologies for Food Safety in Asia” among academia,

Thai business owners and experts from seven countries. Aside from that the networking

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created through this seminar will be carried on and result in further effective cooperation

in this area for food safety in Asia.

May I express my sincere appreciation to the Organizing Committee from both FFTC and

Kasetsart University who has provided great contribution and efforts to prepare necessary

arrangement for this workshop. My special thanks also go, especially, to Dr.Takashi

Nagai, Deputy Director of FFTC, FFTC organizing committee, honorable speakers and

all participants for your devotion to spend time in this gathering. Your contribution

obviously has borne the fruits of success to this event.

Once again, may I welcome all honorary speakers from nine countries and from Thailand

as well as all participants. I wish you a very pleasant stay with memorable experiences

both in academic realm and cultural encounter during time with us in Sampran, Nakorn

Pathom

May I officially declare the seminar open and wish you all a great success in this seminar.

Thank you.

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OPENING ADDRESS

Dr. Takashi Nagai


Assistant Professor and FFTC TAC member Dr. Siree Chaiseri, Assistant Professor Dr.

Warapa, our keynote speaker, Dr. Yi-Jen Liaw, distinguished speakers and agrochemical

scientists from different countries, colleagues in the agriculture industry, friends, ladies

and gentlemen, a pleasant good morning to all of you.

On behalf of the staff and management of FFTC, I cheerfully welcome everyone to this

very timely and relevant international workshop on “Risk Management on

Agrochemicals through Novel Technologies for Food Safety in Asia.”

As everyone knows, food safety has long been one of the most important battle cries of

every government in this planet. It is one of the priorities of big international

organizations like the FAO and the World Health Organization. With the alarming rise of

use and misuse of agrochemicals in our fresh foods, scientists and experts need to work

together not just to improve on their research efforts, but more so to explain to our

consumers that judicious use of agrochemicals is important and that there are more eco-

friendly ways of fighting pests and diseases in plants.

We have partnered with Kasetsart University in this endeavor and came up with a

workshop that hopes to bring together community leaders, scientists and policymakers to

promote and enhance program collaborations to effectively address regional, national and

local responses in minimizing the use of chemicals in crop production. We also aim to

create an active network of people who will help in the assessment and monitoring of

agrochemical utilization and organic farming in the Asian Pacific region.

We have gathered a pool of experts from different countries to facilitate a healthy

exchange of information regarding knowledge on novel technologies which our small-

scale farmers in the region can adapt in the future.

It is therefore my sincere wish that for the next three days, our workshop participants will

be able to maximize their interactions so that they will be able to share whatever

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information they have gathered to their extension workers and those who work directly

with fruit and vegetable farmers.

I would like to extend my congratulations to our partners from Kasetsart University for

making this event possible. First, I’d like to thank Dr. Chaiseri and Dr. Warapa, for

taking the lead and worked hard to efficiently mount this well-organized workshop.

Second, let us a give a warm round of applause to all the members of the secretariat for a

job well-done. Thank you for your hospitality and for your warm accommodations.

And to all our speakers, participants and observers, I wish you all a fruitful, productive

and successful workshop. Let us all make it a point to get to know one another and learn

from each other because this is one of the ways where we can really maximize our

interaction and achieve our objectives. Lastly, let us bask in the beauty of this country.

May we all enjoy our brief stay in Thailand!

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Workshop Schedule

FFTC-KU Risk Management on Agrochemicals through Novel Technologies for Food Safety in Asia

November 11-12, 2015

Sampran Riverside, Nakorn Pathom, Thailand November 11, 2015 (Wednesday)

08.00-09.00 Registration for participants

09.00-10.00 Opening Ceremony Moderator: Dr.Kullanart Tongkhao

Welcome Address Assist.Prof.Dr. Warapa Manakarnchanakul Head of Project, Kasetsart University, Thailand

Opening Message Dr. Takashi Nagai Deputy Director, Food and Fertilizer Technology Center for the Asian and Pacific Region

Assoc.Prof.Dr. Siree Chaiseri Interim Vice President for Research Kasetsart University, Thailand

Group Photo

10.00-10.20 Tea/Coffee Break

10.20-11.10 Keynote Speaker: The Techno Rice: An Industrial application of supercritical fluid technology in Taiwan

Dr. Yi-Jen Liaw Senior Technical Advisor, Taiwan Quality Food Association, Taiwan

Session I Overview: Risk assessment and risk management for controlling and limiting agrochemicals Chairperson: Dr.Yi-Jen Liaw

11.10-11.40 Analysis and risk assessment of pesticides in Korean agricultural products

Dr.Kwang-Geun Lee Professor, Department of Food Science and Biotechnology, Dongguk University-Seoul, Korea

11.40-12.10 Current situation of pesticides use in Indonesian agricultural products

Ms. Anna Rahmianna Researcher, Indonesian Legumes and Tuber Crops Institute, Indonesia

12.10-12.20 Q&A for Session I

12.20-13.30 Lunch Break

13.30-14.00 Current use of pesticides in the agricultural products of Cambodia

Dr. Preap Visarto Director, Plant Protection sanitary and Phytosanitary Department, Cambodia

14.00-14.30 Pesticide residues in food and the environment in the Philippines: risk assessment and management

Ms. Cristina Bajet Scientist I/University Researcher III, National Crop Protection Center, Crop Protection Cluster, U.P. Los Banos,

https://fftcku.files.wordpress.com/2015/09/logo2.jpg

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Philippine

14.30-14.45 Q&A for Session I


Session II Novel technologies in practical: Agrochemicals residues reduction and food safety improvement Chairperson: Dr.Tan Guan Huat

14.45-15.15 Application of ultrasound technology for agricultural product improvement and environmental renovation

Dr. Nomura Nakao Associate Professor, School of life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan

15.15-15.45 Using sanitizer and fine bubble technologies to enhance food safety

Dr. Warapa Manakarnchanakul Assistant Professor, Department of Food Science and Technology, Kasetsart University, Thailand

15.45-16.00 Q&A for Session II

18.00-21.00 Welcome dinner at Phae Phin Thong Restaurant, Nakorn Pathom, Thailand

November 12, 2015 (Thursday) Moderator: Dr.Kriskamol Na-Jom

Session III Analytical approaches for agrochemical determinations

Chairperson: Dr. Kwang-Geun Lee

9.00-9.30 Application of Non-Target Analysis by High-Resolution Mass Spectrometry

Dr. Chung-Huang Wang

Food Industry Research and Development Institute (FIRDI), Product and Process Research Center, Taiwan

9.30-10.00 Chemometric approach to the

optimization of HS-SPME/GC-MS for

the determination of multiclass

pesticide residues in fruits and

vegetables

Dr.Tan Guan Huat

Professor, Department of Chemistry,

Faculty of Science, University of Malaya,

Kuala Lumpur, Malaysia

10.00-10.15 Q&A for Session III


Session IV Key success for Good Agricultural Practices (GAP)

Chairperson: Dr.Warapa Mahakarnchanakul

10.30-11.00 Thai good agricultural practice (GAP) Dr. Rungnapa Korpraditskul

Head, Kasetsart University Research and Development Institute (RDAC), Kamphaeng Saen Campus, Nakhon Pathom, Thailand

11.00-11.30 Development of good agricultural practice (GAPs) model for tea, rice and vegetable in Vietnam

Dr. Dao Bach Khao

Deputy Head, Pesticide, Weed and Environment Division, Plant Protection

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Research Institute (PPRI), Vietnam Academy of Agricultural Sciences (VAAS), Vietnam

11.30-12.00 Producers’ perceptions of public Good Agricultural Practices and their pesticide Use: the case of Durian farming in Pahang, Malaysia

Dr. Yuichiro Amekawa

Assistant Professor, International Course Coordinator, School of Agriculture, Kyushu University

12.00-12.15 Q&A for Session IV

12.15-13.30 Lunch Break

13.30-14.00 Successful case: Key success factor and constraints of control chemicals production system

Dr. Chatchawal Telavanich

Owner, Chatchawal Orchid Co., Ltd.

14.00-14.30 Successful case: Key success factor and constraints of organic production system

Mr. Arus Nawarach

Director, Sampran Organic Farm Model, Nakhon Pathom, Thailand

14.30-16.00 Group Discussion

16.00-16.20 Closing Session Assist.Prof.Dr. Warapa Manakarnchanakul

Head of Project, Kasetsart University, Thailand

Dr. Takashi Nagai


18.00-21.00 Welcome dinner at Jinda Cabana Restaurant, Nakorn Pathom, Thailand

Note: The schedule may be subjected to change if necessary

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SPEAKER BIODATA

Yigen Liaw, PhD

Office Address: Taiwan Quality Food (TQF) Association

6F. No. 219, Section 4, Chungu Hsiao East Road, DaAn

District, Taipei 10690, Taiwan, ROC

E-mail: [email protected]

Professional Experience

Period Position Organization

2012/6 – 2013/11 Director Wei Chuan Food Co. Taiwan

2014/2 – 2015/6 Secretary Taiwan Food GM

General

2015/6 – present Senior Technical Advisor Taiwan Quality Food

Kwang-Geun Lee, PhD

Office Address: Department of Food Sci & Biotech,

Dongguk Univeristy, 32 Donggukro, Ilsandong-gu,

Goyang, Korea

Tel: 82-31-961-5142 and 82-10-2023-6026 Fax: 82-31-

961-5183



Period Position Specialty Organization

2001 – 2002 Lecturer toxicant analysis University of California, Davis

.

2002- present Professor Dongguk University

2007- present Director Dongguk Business Incubation

Center

mailto:[email protected]

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Rahmianna Anna, PhD

Main field of expertise: aflatoxin contamination

managing environmental (soil moisture)


Visarto Preap, PhD

Main field of expertise: crop protection, bio-security,

Plant Quarantine management and insect ecology & IPM

Tel: 855 (011) 622 916




Current Research Scientist Director Plant Protection Sanitary

and Phytosanitary Department

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Bajet Cristina M., PhD

Office Address: National Crop Protection Center, Crop

Protection Cluster, U.P. Los Los Banos




Present Scientist University Researcher III

Nakao Nomura, PhD

Main field of expertise: Animal Cell Culture Technology,

Waste Water Treatment, Shrimp Aquaculture




2002.5 – 2003.10 Visiting Researcher University of Philippines Los Banos

2003.10 – 2010.7 Assistant Prof essor University of Tsukuba

2010.7 - Present Associate Professor University of Tsukuba

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Mahakarnchanakul Warapa, PhD

Main field of expertise: Mycotoxin detection in food

products, GMP/HACCP system, Safety of minimally

processed procedure, Microbial Stress Response

Office Address: Department of Food Science and

Technology, Faculty of Agro-Industry, Kasetsart University

50 Ngamwongwarn Rd. Ladyao, Jatujuk, Bangkok, 10900,

Thailand

Tel: +66-2562-5036


Chung-Huang Wang, PhD

Office Address: Food Industry Research and Development

Institute, Hsinchu, Taiwan


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Prof. Dr. Tan Guan Huat, PhD

Main field of expertise: Chemical Analysis

Office Address: University of Malaya.




2000-present Examiner Institut Kimia Malaysia

2000 – present Admissions’ committee Malaysia Institute of Chemistry

Member

2012- present Co-Director ANAC

Korpraditskul Roongnapa,PhD

Main field of expertise: Microbial and Pesticide

contamination in Fruit and vegetable, Farm Waste

Utilization and Environmental Management, Pesticide

degradation, Good Agricultural Practices; Inspection and

Certification System, Agricultural Commodities Standard

setting, and Public and private standard ThaiGAP Institute

Board Committee

Office Address: 3rd

Floor, University Building Center

Kasetsart University,Kamphaengsaen, Nakhon Pathom,

73140, Thailand


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Amekawa Yuichiro, PhD

Office Address: Kyushu University 6-10-1 Hakozaki

Higashi-ku Fukuoka 812-8581 Japan

Tel: +81 92 642 7178


DAO BACH KHOA, PhD

Main field of expertise: Insect physiology and biology

molecular including Digestion System, Isolation and

Bioassay of Natural Products as Pesticides. And Pesticide

Science, Pesticide Residue.

Office Address: Plant Protection Research Institute

(PPRI) – Dong Ngac – Tu Liem – Ha Noi

Under: Vietnam Academy of Agricultural Sciences

(VAAS)

Tel: 84-90-6290573 Fax: 84-43-8363563



Period Position Specialty Organization

2006 – 2007 Researcher Plant Protection Dong Ngac-Tu Lium-Ha Noi

Research Institute

2007-2012 Master&PhD Kobe University

2012-present Head of Department Dong Ngac-Tu Lium-Ha Noi

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PAPER PRESENTATION

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THE TECHNO RICE: AN INDUSTRIAL APPLICATION OF

SUPERCRITICAL FLUID TECHNOLOGY IN TAIWAN

Yi-Jen Liaw

Senior Technical Advisor, Taiwan Quality Food (TQF) Association

6F., No. 219, Section 4, Chungu Hsiao East Road,

DaAn District, Taipei 10690, Taiwan

Email: [email protected].

ABSTRACT

Techno Rice, which is the rice treated with the state of the art supercritical carbon dioxide

rice cleaning process, is tasty, clean, safe, and environmentally sound. As for the preparation of

boiled brown rice, no additional rinsing and presoaking is needed and requires much less

cooking time as well. In the meantime, it retains the fragrant and savory characters of rice with

added remuneration of dietary fiber, antioxidants, and polyols which include: ferulic acid, its

esterified derivatives (oryzanols), tocopherols and other phenolic compounds. The result shows

that the partially defatted rice is more palatable and healthier, and it has longer shelf life.

Furthermore, the process can remove pesticides residues, reduce microbe content and destroy

existing insect and insect eggs to make consumption of the rice safer than ever. Techno Rice is

tasty, healthy, safe and surely an alternative to compete with other products in the global rice

market.

Keywords: Supercritical Carbon Dioxide, Techno Rice, Brown Rice

INDTRODUCTION

Supercritical fluid technology has been applied in industries for more than twenty years. Its

first industrial application was on the extraction of hop resin, and in the1970’s it was used to

decaffeinate coffee beans in Europe. Although there is considerably high number of works being

tested in various applications, in the end of twentieth century, the supercritical fluid technology

does not become the prospective industrial process yet as it was expected. Nevertheless, in the

twenty-first century, it has been considered as one of the “Green Chemistry” processes. Unless it

can be widely accepted by the industry, the very growth of this environmentally friendly clean

process will be in jeopardy.

The research on this new technology in Taiwan is still insignificant in terms of governmental

support and number of the researchers involved. Most of work adopting this technology focused


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on new applications in laboratory scale. The design of industrial apparatus and commercial

process plant for supercritical fluid technology is rare. The first commercial plant for using

supercritical carbon dioxide as cleaning solvent for rice, denoted as “Techno Rice”, was

commissioned successfully in Taiwan in the end of 1999. It can be considered as the stepping-

stone for the industrial application of the supercritical fluid technology in Taiwan.

Supercritical fluids extraction (SFE)

Supercritical fluids extraction, in most case, uses carbon dioxide as solvent and conducts

extraction under its supercritical condition (McHugh, M. A., Krukonis, V. J., 1986). It contains

all benefits of gaseous and liquid solvents, which are used in traditional chemical engineering

unit operations. Carbon dioxide is non-toxic; it does not leave any residual side effects that may

harm human bodies as other organic solvents may do. Supercritical carbon dioxide is a fluid in

between the status of gas and liquid. It has high diffusivity, low viscosity, and non-flammable,

and does not pollute and has high selectivity. It uses differences in temperature and pressure to

adjust its fluid characteristics to help extraction technology becomes more flexible.

Basically, supercritical carbon dioxide technology raises the temperature and pressure of

carbon dioxide to above critical level, utilizes its liquid-like solubility, gas-like diffusivity,

viscosity and surface tension to expedite the extraction process. In other words, under

supercritical conditions, carbon dioxide is able to penetrate into the solid matrix as gas and

dissolve impurities in and on the matrix as liquid while “laundering” the rice.

In addition, supercritical carbon dioxide can be separated from extract easily, without having

the solvent recovery by boiling up the washed-out organic solvent as a practice for traditional

liquid extraction process. Therefore, it leaves no organic solvent residue, as we may consider the

process as a dry cleaning process for the rice, and will not pollute environment in any way. SFE

process fits perfectly into the basic principle for modern green processing technology.

Starting from the 60’s, many research groups, primarily in Europe and then later in the U.S.,

have examined SFE for “advance” extraction processes. European researchers emphasized

extraction from botanical substrates, for example, spices, herbs, coffee, tea, and so on, uses

predominantly supercritical carbon dioxide. By the 80’s there are several large SFE process

plants in operation in Germany, UK and US, for decaffeinating coffee and tea and extracting

flavors and essential oils from hops, spices, and herbs (Lack and Simandi, 2001.)

Ever since the launch of this new technology, the focus of supercritical fluids is on water,

carbon dioxide and alcohol, because those are considered as green and sustainable in nature. It

comes with no surprise that one application attempts to use supercritical carbon dioxide as a

cleaning solvent to wash out the contaminants on a solid matrix. This is a sensational idea;

however, would it worth using rather expensive technology on low-tech industry like rice milling,

which has been considered cheap?

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Supercritical fluid rice cleaning process (SFRCP)

This is a simple idea but the technology itself is not an easy task. Supercritical fluid rice cleaning

process is a typical SFE practice emulating the pioneer SFE decaffeinating process that has

existed for quite some time in coffee and tea industries. Generally speaking, supercritical fluid

rice process (SFRCP) uses carbon dioxide under supercritical conditions to permeate into rice

and dissolve possible pollutants from the surface and within (Lack and Simandi, 2001). SFRCP

can also exterminate existing germs and insect eggs so that the rice is much cleaner and safer for

human consumption.

The process is constructed with three 5.8 cubic meter extractors which operate cascades in

series. Like most basic laboratory SFE unit, the main engineering design of this commercial

apparatus consists of extractor, separator, pressure pump and heat exchanger (Fig. 1). In addition,

a co-solvent pump is equipped for modification of polarity of the extraction solvent and

supercritical carbon dioxide in this case.

A hermetic centrifugal pump is used as a booster pump to avoid cavitation to replace pre-

chill water bath, which is a common practice in laboratory unit, for the sake of efficiency and

spatial design. A 450 horsepower plunger pump, tandem with the booster pump, is capable of

delivering 20,000 kg/hr. of liquid carbon dioxide and pressurizing operation pressure up to 350

bars (Fig. 2). An inclined tube bundle heat exchanger is used to heat up the pressurized liquid

carbon dioxide to reach its critical temperature before entering the extraction vessel.

For recycling of the carbon dioxide at gaseous phase, after evaporation at the separator by

means of pressure reduction and temperature increment, another tube bundle heat exchanger is

used as an inline condenser during operation.

Carbon dioxide in the pressure vessel is then equilibrating with storage tank (working tank)

which stores liquid carbon dioxide under its equilibrium vapor at temperature around 20 degree

Celsius. A two-stage compressor is used at the final step for carbon dioxide recovery before

opening of the extraction vessel. The compressor can deplete the gaseous carbon dioxide in the

extraction vessel to 4.5 bars or lower to minimize carbon dioxide losses. As a result, the

empirical data for average recovery of carbon dioxide is approximately 99.5%.

Co-solvent is introduced into the process by mean of a diaphragm pump before pressurized

liquid carbon dioxide entering the pre-heater. Presumably, the surface area and the contacting

time between carbon dioxide and co-solvent are sufficient to reach equilibrium and to become a

single phase, by definition.

It is crucial to avoid any direct contact of rice with water, as it may present or precipitate

inside the vessel as a second phase. Monitoring the stroke of the co-solvent pump and tuning the

concentration of the co-solvent, SFRCP can further manipulate the moisture content of the rice.

In the meantime, the shrinkage of the rice kernel must be prevented because carbon dioxide may

take out the moisture from the rice and diminish the appearance as well.

Automatic controlling system, quick-acting closure design, safety sensor and interlock

system have made the four-story SFE harboring operation plant which requires a minimum of

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two operators to keep it fully functional (Fig. 3). Extra precaution has been taken with a safety

measure in mind; therefore, a touch screen control panel has been installed alongside the top and

the bottom quick-acting closure for onsite operation and supervision (Fig. 4).

SFRCP is operated semi-continuously with three extraction vessels arranged in sequence.

The supercritical carbon dioxide, with or without co-solvent, is entering the vessel from the

bottom continuously. The three extraction vessels normally will operate in cascade mode, which

can be divided into two different time periods, namely extraction time and down time. In doing

so, we can save up to 1/3 of process time, in view of a single batch, by partially overlapping the

extraction time period while two vessels are operated as a stringed extractors. In other word,

while one vessel operates with high-pressure carbon dioxide the other two vessels are either

depressurizing or ready for unloading and loading, or pressurizing and ready for in tandem with

the existing vessel.

After the extraction, supercritical carbon dioxide can then be gasified by means of

depressurization and evaporation in the separator. As the supercritical carbon dioxide has

rendered its solvent power to gas phase, the water in oil emulsion precipitated in the separator

can then accumulate and be drained out for further separation, which can be done easily by

breaking up the emulsion with steam. Gas carbon dioxide is subsequently introduced into a

washing column to strip away trace residue of contaminant that may be carried over before it is

re-condensed into liquid form and drained down to the working tank.

The washing column is basically a countercurrent extraction packed column. To avoid

overflow and for easy handling, the gas carbon dioxide, as continue phase, is ascending from the

bottom and the hot water, as disperse phase, is dripping down from the top. Water level of the

column is monitored automatically for depleting and replenishing.

Gas carbon dioxide escaped from the washing column and saturated with water is introduced

into the condenser to conclude with the “laundering” cycle. Condensed water occurs as it passes

through condenser and is collected from the bottom of the working tank; which is designed with

10-degree incline, and drains out automatically with a sensor monitoring level gauge.

The spent water, which is saturated with carbon dioxide, is introduced into a gas/liquid

separator to deplete the gas carbon dioxide before coming into contact with micro filters and

active carbon for further purification.

The hermetic centrifugal pump is designed to compress the liquid carbon dioxide effectively

to about 14 bars higher than what it is in the working tank, which is equilibrated with the

pressure of separator at around 55 bars. Theoretically, the centrifugal pump can be used directly

as the pressure pump for the process. In fact, it is even more suitable than plunger pump for

delivering massive liquid flow. However, it is never realistically feasible, as the pump has

reached its limitation in terms of the diameter of the impeller and the length of shaft inside the

stage casing.

Relatively speaking, plunger pump is still the number one choice for the industrialized

commercial high-pressure process (Vetter et al., 2001). In practical, the reciprocal pump featured

24

with three plungers and equipped with dampers to reduce pulsation is commonly used for most

of the existing commercial SFE processes.

SFRCP has integrated seamless with the traditional rice milling process at a production

capacity of 2 to 4 metric tons of rice per hour (Fig. 5). The utilities for the complete process have

included air compressor, steam boiler and chiller, cooling tower, water treatment and rice

transport system. Together with the main devices of the SFRCP the total electric energy

requirement for the plant is around 1700kw. In other word, it can never be considered as an

energy saving process if compared to traditional rice milling.

The techno rice

Techno Rice is named in comparison to organic rice, in a sense that the using of modern

technology to ensure the treated rice has its quality assured like organic rice. Techno Rice has

made the nutritious brown rice more palatable. It is worth mentioning that, the Techno Rice

offers a greener, safer and technologically sound alternative, which gives consumers full peace

of mind.

Supercritical fluid extraction itself can be called clean process or green technology. This

technology reduces contaminants, such as pesticides, microorganisms and pests in the treated

rice products to the minimum, retaining all the nutrients in the rice. SFRCP has changed the

undesirable traits for brown rice those are not easy to store, cook, and roughness in eating.

Techno Rice is aromatic, chewy, and leaving no coarsely mouth feel. As consumers become

more aware of the food, they dwell on issues like nutraceuticals, functional ingredients,

convenience, and safety; Techno Rice, especially brown rice, would be an excellent product to

target this sophisticated market.

When it comes to a basic fact that although white rice is indeed tastier than brown rice, it

comprises only a compact agglomeration of starch granule. Even though people are aware of

nutritional values, nutraceuticals and functional ingredients in the brown rice, the pericarp, seed

coat and nucellus that make up the caryopsis coat (bran) in brown rice have kept consumers

away from being rational.

Rice bran contains most of the oil in the rice and covers the outer skin of rice grains with a

waxy layer, hampering water absorption of the rice. To cook the rice, the starch granules need to

absorb sufficient amount of water in order to swell up and rupture for the carbohydrate to be

fully gelatinized. With the formation of the complex between free fatty acids, carbohydrates and

protein plus the deferring of water absorption by its natural barrier, the rice has been referred to

be poor in eating quality after cooking.

Brown rice is more susceptible to deterioration than white rice. Lipase, an extremely active

enzyme underneath the bran, can hydrolyze rice lipid into free fatty acid as soon as the husking

process begins. Free fatty acid can be oxidized to hydroperoxides and other smelly carbonyl

25

compounds; which will produce off-flavor and form complex with starch and protein and

accelerate the aging process of rice.

Techno Rice offers a simple solution for the problem by taking away deterioration elements

as such. Supercritical carbon dioxide removes the waxy layer and fatty acid inside rice bran,

prohibits the formation of polymer complex and prevents rice from aging.

Once the odor caused by oxidized fatty acid is removed, the genuine aroma of cooked rice

can be released. In addition, SFRCP causes micro-fracture on the bran; consequently, Techno

Rice can absorb water much more effectively. The soaking time can then be shortened and the

process to swell and gelatinize starch during cooking can be expedited.

The result is a nutritious tasty brown rice, which is soft inside and chewy on the outer layer.

It has been shown that SFRCP treated brown rice can be prepared similar to white rice without

the need for overnight soaking which is a common practice in Taiwan. Apart from grainy texture,

the outer skin has become chewier; the perception for the SFRCP brown rice is just as tasty and

soft on texture as regular white rice. The average cooking time can be cut short by approximately

25% as well.

Similar to the carbon dioxide packaging, which is widely used for perishable dairy and meat

products and rice storage for military usage as well, Techno Rice can sustain this practice as an

additional benefit by a designed one-way valve on the bag (Fig. 6). As the chemical potential of the

dissolved carbon dioxide in Techno Rice is much higher than that of its surrounding air, the carbon

dioxide will gradually be released from the rice matrix.

Like what is commonly used in coffee bean packaging, a one-way valve has provided a conduit

for the excess carbon dioxide to escape. Being different from the carbon dioxide that generates slowly

in the coffee bean during storage, an enormous amount of carbon dioxide that has previously

dissolved in the rice during SFRCP busts out and pops. In addition, the predominant carbon dioxide

can expel the air in the bag and provide a strictly anaerobic storage environment for the rice. Since

carbon dioxide can again dissolve back into the solid matrix and water in rice, after an empirical time

period of 7 to 14 days, the package will then shrink automatically. It creates something similar to a

vacuum package and results in an even better preservation system than traditional gas package or

vacuum package alone.

The concentrated carbon dioxide dissolved in rice can produce carbonic acid, which can further

reduce the pH value of rice to around 3, theoretically. Such acidic condition can further prohibit

germination of microbes and pest eggs. Although the rice can never be completely sterilized by the

process hitherto used, most of those hidden bio-contaminants can be further inhibited by the presence

of carbon dioxide. Therefore, Techno Rice can be stored much longer, and even the most pest-prone

brown rice requires no needs for commercial vacuum packaging.

CONCLUSION

With mounting pressure from the WTO to open market, rice industries in Taiwan are adapting

the new technology to revive and fend them from the pending competition. Organic rice can

26

certainly increase farmers’ income, as well as, provide consumers with a healthier and safer

option for rice. Due to the facts that organic rice production is very labor intensive and has high

production cost and other various factors, it has never been able to increase significantly and

reach the magnitude as what has been anticipated. Furthermore, subtropical climate and over

cultivated rice field have made it even more difficult for the farmer to achieve the organic

farming criteria while maintaining commercially feasible production cost to compete globally.

The partially defatted rice, namely Techno Rice, is more palatable, healthier and has longer

shelf life. The process can removes pesticides residues, reduces microbe content and destroy

existing insect and insect eggs to make consumption of the rice safer than ever.

Techno Rice is tasty, healthy and safe and surely is an alternative to compete with variety of

rice in the global rice market. It is commercially available in the Taiwan for the past fifteen years.

How well this product can survive in the traditional rice market, locally as well as internationally,

is still too early to determine. Nevertheless, it has exploring a new application of supercritical

fluids technology on staple food for the Homo sapiens.

REFERENCES

Arai, Y., Sako, T., and Takebayashi, Y. (Eds.) 2002. “Supercritical Fluids – Molecular

Interactions, Physical Properties, and New Applications.” Springer-Verlag, Berlin

Heidelberg.

Lack, E. and Simandi, B. 2001. Supercritical Fluid Extraction and Fraction from Solid Materials.

In “High Pressure Process Technology: Fundamental and Applications,” eds. A. Bertucco,

and G. Vetter, pp 537 – 575. Elsevier Science B. V., Amsterdam.

McHugh, M. A. and Krukonis, V. J. 1986. “Supercritical Fluid Extraction – Principles and

Practical.” Butterworths Publishers, Boston.

Stahl, E., Quirin, K. W., and Gerard, D. 1988. “Dense Gases for Extraction and Refining,”

Springer-Verlag, Berlin Heidelberg.

Vetter, G., Luft, G., and Maier, S.. 2001. Design and construction of high-pressure equipment for

research and production. Chapter 4 in “High Pressure Process Technology: Fundamental

and Applications,” ed. A. Bertucco, and G. Vetter, pp 537 – 575. Elsevier Science B. V.,

Amsterdam.

27

Fig.1. 3D drawing of supercritical carbon dioxide rice cleaning system (SFRCP)

Extractor

Separator

Condenser

Pump Co2 Tank

28

Fig. 2. High-pressure plunger pump equipped with dampers

29

Fig. 3. Quick-acting closure at the top of the extraction vessel

30

Fig. 4. Quick-acting closure at the bottom of the extraction vessel

31

Fig. 5. Overview of SFRCP apparatus with three vessels arranged in series

32

Fig. 6. Commercial SFRCP rice (Techno Rice) package with one-way valve.

33

Session I : Overview: Risk assessment and risk management for

controlling and limiting agrochemicals

34

ANALYSIS AND RISK ASSESSMENT OF PESTICIDES IN KOREAN

AGRICULTURAL PRODUCTS

Kwang-Geun Lee

Department of Food Science and Biotechnology, Dongguk University-Seoul, 32, Dongguk-ro,

Ilsandong-gu, Goyang-si, Gyeonggi-do, 400-820, Republic of Korea

Phone: 82-31-961-5142, E-mail: [email protected]

ABSTRACT

The objective of this study was to establish an analytical method to measure pesticides and to

analyze pesticide residue levels of Korean agricultural products such as yuza (Citrus junos Sieb.

ex Tanaka), yuza tea, and ginseng products. Risk assessments were also performed by

calculating estimated daily intake (EDI) and acceptable daily intake (ADI). In addition, kinetic

study such as the degradation order of pesticides in yuza tea was carried out. Acequinocyl,

spirodiclofen and carbendazim were detected in yuza samples in the concentration range of

0.07–0.15 µg/g, 0.11–1.89 µg/g, and 0.03–5.15 µg /g, respectively, whereas chlorpyrifos,

prothiofos, phosalone, and deltamethrin were not detected in yuza or yuza tea. The

concentrations of acequinocyl, spirodiclofen and carbendazim ranged from 0.18–1.05 µg/g,

0.13–0.29 µg/g, and 0.17–2.36 µg/g, respectively, in yuza tea samples. The percent ratios of EDI

to ADI for acequinocyl, spirodiclofen, and carbendazim were 24.6%, 22.7%, and 58.5%,

respectively. The degradation order of the seven pesticides in yuza tea was as follows:

acequinocyl > chlorpyrifos > spirodiclofen > carbendazim > deltamethrin > phosalone,

prothiofos. For the accurate analysis of Korean ginseng products a new analytical method was

developed based on gas chromatography-triple quadrupole tandem mass spectrometry (GC-

MS/MS). The analytical method was validated and the most frequently detected pesticide was

tolclofos-methyl. Tolclofos-methyl was detected in 86.4 % of fresh ginseng (18.25-404.5 ㎍/kg),

91.7 % of red ginseng (13.14-119.4 ㎍/kg), and 87.5 % of dried ginseng (23.15-3673 ㎍/kg).

Keywords: Pesticide; Residue; Yuza; Yuza tea; Risk assessment; Ginseng; Validation; GC-MS/MS

35

INTRODUCTION

Yuza, Citrus junos Sieb. ex Tanaka, is a citrus fruit harvested in Korea, China, and Japan.

Particularly in Korea, the fruit is commonly processed into beverages and herbal medicines due

to its special flavor and effectiveness against colds. Most parts of yuza fruit such as the peel,

juice, and seeds have been used (Lan-Phi et al., 2009). The beneficial health effects of flavonoids

in yuza such as antioxidant, anti-carcinogenic, anti-viral, and anti-inflammatory activities have

been reported (Fujihara and Shimizu, 2003; Yoo et al., 2004).

Ginseng (Panax ginseng C.A. Mayer) is a valuable herb that has been used extensively in

Eastern Asian countries, such as Korea, China, and Japan, for more than 5,000 years. Ginseng as

a medicine has been studied thoroughly. Ginsenosides are major components having

pharmacological and biological activities, including immune, cardiovascular, central nervous

system, endocrine, anti-diabetic, anti-tumor, and antioxidant activities (Attele et al., 1999).

Ginseng is generally harvested after a 5- or 6-year cultivation period, or even a 10-year period.

Such a long cultivation period combined with the widespread use of pesticides in ginseng

production has led to the presence of pesticide residue in ginseng (Kim and Lee, 2002).

A number of pesticides have been widely used for pest control in crops and fruits at various

stages of cultivation. Pesticides are used for better yields and quality during post-harvest storage.

However, pesticide residues are of concern, because these substances are potential health hazards

(Taylor et al., 2002). In particular, pesticide residues may persist in plant tissues and appear in

the pulp and juice of fruits and vegetables. Determining pesticide concentrations is important, as

people who eat relatively large quantities of these foods are more at risk than others. In Korea,

the pesticides authorized by Korea Crop Protection Association for yuza production are

chlorpyrifos, prothiofos, phosalone, deltamethrin, acequinocyl, spirodiclofen and carbendazim

(Korea Crop Protection Association, 2009). Pesticide residue monitoring of fruits and vegetables

has been conducted to confirm the proper use and exact concentrations of pesticides. Maximum

residue limits (MRLs) have been established for agricultural products in many countries to avoid

the health hazard caused by pesticide residues. Health safety limits for human health are

typically expressed as acceptable daily intake (ADI). The standard method to evaluate human

exposure is based on the average consumption per person per day, Korean average adult weight,

and pesticide residue data (Park et al. 2010).

Tandem mass spectrometry (MS/MS) provides a much higher degree of assurance in the

identification of an analyte than any other single stage mass spectrometry technique. One of the

advantages of MS/MS is to increase S/N ratio and selectivity. Due to the power of MS/MS, the

confirmation of target compounds can be achieved with a higher level of confidence. Among the

various mass analyzers that can perform tandem mass spectrometry, triple quadrupole mass

spectrometers have recently been recommended for the analysis of pesticide residues in crops

(Walorczyk, 2007).

Although many scientists have analyzed pesticide residues in various fruits, the analysis of

pesticides in yuza has been carried out a few. Yamamoto et al. (1982) reported acaricide residue

on yuza fruits grown in vinyl houses. Goto et al. (2003) reported on a simple and rapid method to

36

identify pesticides in citrus fruits by electro-spray ionization tandem mass spectrometry. Micro-

extraction procedures were compared to determine pesticides in oranges using liquid

chromatography–mass spectrometry by Blasco et al. (2002).

The objective of this study was to establish an analytical method for seven pesticides often

used for yuza and to analyze pesticide residue levels in yuza and yuza tea produced in Goheung,

Korea. A risk assessment of pesticides in yuza tea was also conducted by determining the

estimated daily intake (EDI) and ADI. In addition, the kinetic parameters for the degradation of

pesticides were investigated. Regarding to the ginseng, a robust multiresidue method for the

analysis of 32 pesticides of different classes was studied using GC/MS/MS.

MATERIALS AND METHOD Chemicals

Pesticides,acequinocyl, chlorpyrifos, spirodiclofen, carbendazim, deltamethrin, phosalone, and

prothiofos, were purchased from Sigma-Aldrich (Germany) and stored at room temperature.

Sampling and Storage

Yuza sampling was conducted from October to November 2009. Yuza samples, grown in

Goheung, Korea, were placed in polyethylene bags and transported to the laboratory immediately

after harvest and stored at -20°C. Yuza teas samples (n = 25) were collected from Hansung Food

Inc., Goheung, Korea during 2009 and 2010. Ginseng samples (fresh ginseng, red ginseng, dried

ginseng) were obtained from local ginseng agricultural cooperative federations located in Seoul.

All samples were produced in South Korea. Samples were analyzed within 24h and stored at 4℃

until the moment of extraction. No degradation of pesticides was detected under the storage

conditions.

Extraction and purification

Extraction and purification of various pesticides were carried out by the previously published

papers (Lee and Lee, 2012; Lee and Jo, 2012; Nam et al., 2015).

Analytical method

Gas chromatography (GC) analyses were carried out on an Agilent Technologies Model 6890N

gas chromatograph with nitrogen phosphorus detector (NPD) and auto-sampler (Model 7683).

Data acquisition and analysis were performed with the G1035 Wiley Library. Separation was

performed on a DB-5 capillary column (30 m × 0.250 mm i.d × 0.25 µm film thickness, Agilent

Technologies). Injection temperature was held at 260°C. The injection mode and volume were

split mode and 1 µl, respectively. Helium was used as the carrier gas at a flow rate of 1.0 ml/min.

37

The oven temperature program was initially 120°C (hold 2 min), increased to 220°C at

10°C/min, to 250°C at 7°C /min, and then to 280°C at 7°C /min (hold 15 min). The NPD

temperature was maintained at 280°C.

Gas chromatography/mass spectroscopy (GC/MS) analyses were carried out on an Agilent

Technologies Model 6890N gas chromatograph with a mass selective detector (Model 5975,

Agilent Technologies). Data acquisition and analysis were performed with the Chemstation

(Agilent Technologies). Deltamethrin separation was performed on a DB-5 MS capillary column

(30 m × 0.250 mm i.d × 0.25 µm film thicknesses, Agilent Technologies). Injection temperature

was held at 260°C. The injection port was heated to 260°C, and the splitless injection mode was

used. Injection volume was 2 µl. Helium was used as carrier gas at a flow rate of 1.0 ml/min. The

oven temperature program initially consisted of 120°C (hold 2 min), increased to 220°C at 10°C

/min, to 250°C at 7°C /min, and then to 280°C at 7°C /min (hold 15 min).

The high performance liquid chromatography (HPLC) was carried out on a Waters

Instrument Model 1525 consisting of an auto-sampler (Model 717, Waters) and a photodiode

array detector (Model 2998, Waters). A C18 column (Zorbax Eclipse XDB, Agilent Technologies,

4.6 × 250 mm, 5 µm particle size,) was used for separation at 40°C. The injection volume of the

sample was 10 µl. Acequinocyl separation was carried out with water and acetonitrile (13%:

77%) at a flow rate of 0.8 ml/min for 30 min. The UV absorbance of acequinocyl was monitored

at 250 nm. Separation of sprodiclofen and carbendazim was performed by gradient elution with

water (A) and methanol (B) at a flow rate of 1.0 ml/min. UV absorbance of spirodiclofen and

carbendazim was monitored at 254 nm and 279 nm, respectively. The gradient elution program

was as follows: initial 50% A; 0–45 min, 10% A; and a 45-50 min return to the initial conditions.

Processing the raw chromatographic data and data collection were performed using the Empower

2 Pro program (Waters, Milford, MA, USA).

Pesticide analysis was performed using a Varian 3800 gas chromatograph equipped with an

electronic flow control (EFC) and fitted with a triple quadrupole mass spectrometer (GC-MS/MS)

(Varian Instrument, Sunnyvale, CA, USA). Chromatographic separation was performed on

Varian VF-5ms Columns (30 m × 0.32 mm, 25 μm film, Walnut Creek, CA. USA). Injection

temperature was held at 250oC, and injection volume was 1 μl. The oven temperature was held

for 2 min at 70°C, ramped to 200 °C at 20°C/min, increased to 220°C at 2°C/min, and then held

for 3 min and finally increased to 300℃ (hold 3min) at 10℃/min (total run time equaled 35 min)

with a flow of 1.0 ml/min. Helium (99.999 %) was used as the carrier gas and the linear velocity

was 30 cm/s. Triple quadruple system was operated in electron impact ionization mode (EI,

70 eV). Argon (99.999 %) was used as the collision gas. The dwell time was 0.5 s. The trap,

manifold, and transfer line temperatures were set at 200°C, 40°C, and 280°C, respectively. Split

ratio and split vent valve state were programmed as follows: initial (open, 5:1); 0 min (closed,

off); 1.5 min (open, 100:1); 3.0 min (open, 30:1). The MS/MS system entails two fundamental

steps between the formation and detection of ions. In the first step, the precursor ion or an entire

cluster of parent ions is isolated in the trap, and in the second step, the dissociation of the

precursor ion or ions is performed by collision with an inert gas.

Method validation

Sample preparation was validated in terms of linearity, repeatability, limits of detection (LOD)

and quantification (LOQ), and recovery. The evaluation of linearity was conducted by injecting a

38

solution containing six standard chemicals (0.02, 0.1, 0.5, 1.0, 5.0, and 10.0 µg/ml). All

standards were injected three times (n = 3). LOD was measured as the analyte concentration

based on signal to-noise ratio of 3, and LOQ was defined as 3.3 × LOD. To determine recovery

of pesticide, each pesticide standard solution (10.0 µg/ml) was spiked to the homogenized

sample. The recovery rate was calculated as (pesticide weight in spiked sample–pesticide weight

in unspiked sample) × 100/spiking pesticide weight. The assay procedure was repeated five times,

and relative standard deviation values were obtained within the same day to evaluate intra-day

precision. To evaluate inter-day precision this assay was carried out over five different days.

Experimental design for the kinetic model

Yuza cultivated in the conventional way was sprayed with a mixture of chlorpyrifos, prothiofos,

phosalone, deltamethrin, acequinocyl, spirodiclofen, and carbendazim at the recommended

normal doses with a sprayer. The amounts of 7 pesticides dissolved in 30mL aqueous solution of

the mixture were from 0.003g to 0.03g. Yuza sprayed with 30mL of pesticide mixture was

processed for yuza tea in the laboratory, because we usually consume yuza tea rather than yuza

worldwide. The sprayed yuza was sliced and the seed was separated. Table sugar was added to

the sliced yuza (1:1) and aged for 1 week in the shade. Sampling was conducted after 2 h, and 1,

2, 3, 5, 7, and 10 days from the last pesticide spraying. The weight of yuza did not increase

during the sampling period; thus, the dilution of pesticide residue was not affected by growth.

Kinetic models employed to estimate the half-life of the pesticide residues in yuza tea after

pesticide treatment such as first-order (FO), zero-order (ZO), and second-order (SO) models are

empirical formulas for estimating the correlations between time (t) and concentration (μg g-1

) of

pesticide residue.

RESULTS AND DISCUSSION

Method validation

Seven pesticides (chlorpyrifos, prothiofos, phosalone, deltamethrin, acequinocyl, spirodiclofen,

and carbendazim) were chosen because of their frequent use during yuza cultivation. The

maximum residue levels (MRLs) for yuza are shown in Table 1. The slopes, Correlation

coefficient values, Limit of detection (LOD), and Limit of quantification (LOQ) are also

summarized in Table 1 for the validation study. An excellent linear correlation was observed

between the pesticide concentration and peak areas, with coefficient correlations values of

0.9750–0.9999. Percent recoveries were 80.4–109.9% for all pesticides. The standard deviation

of recovery rate were <6.9%, suggesting that the extraction procedure was suitable for routine

analysis of the targeted pesticide residues. The LOQs of the method were 0.06–0.33 µg/ml. The

intra-day variability was assayed at five replications on the same day. For inter-day variability

the assay was carried out during 5 sequential days to test precision of each pesticide. The relative

standard deviation (RSD) of intra-day and inter-day variability was <15.9% and 16.9%,

respectively (Table 1).

39

Pesticide levels in yuza and yuza tea samples

The results of the pesticide analysis in yuza cultivated by ordinary and environmentally friendly

cultures (2010–2011) and the maximum residue limits (MRLs) are summarized in Table 2.

Prothiofos and deltamethrin were not detected in yuza cultivated by ordinary culture in 2010.

Chlorpyrifos was found at up to 0.108 mg kg−1

in a single sample of yuza cultivated by ordinary

culture. Spirodiclofen revealed a range of contamination of up to 1.889 mg kg−1

in 16 of 50 yuza

plants cultivated by ordinary culture. The 37 yuza cultivated by ordinary culture contained the

highest level of carbendazim at 5.148 mg kg−1

. In 2011, spirodiclofen and carbendazim were

detected at lower than 0.334 and 3.840 mg kg-1

, respectively. The other pesticides were not

detected or quantified. Chlorpyrifos, prothiofos, phosalone, deltamethrin and acequinocyl were

not detected or quantified in yuza cultivated by environmentally friendly culture in 2010.

Spirodiclofen and carbendazim were found in eight of 30 yuza samples at up to 0.801 mg kg−1

and in 13 of 30 yuza samples at up to 2.907 mg kg−1

, respectively. Carbendazim was detected in

eight samples at concentrations ranging from ND to 0.340 mg kg−1

in 2011. The remaining

pesticides were not detected or quantified. Carbendazim was detected in all samples. The

pesticide residues in all yuza samples were lower than the MRLs established by Korean

legislation. However, because of possible health effects and widespread use, continuous

monitoring of carbendazim is necessary in the future. In addition, higher levels of the tested

pesticides were present in yuza samples produced in 2010 than in 2011.

Risk assessment

The results of human exposure to pesticides based on yuza tea intake are shown in Table 3. The

EDIs of acequinocyl, spirodiclofen, and carbendazim were 6.6438 × 10-3

, 3.1733 × 10-3

, and

1.7562 × 10-2

mg/day, respectively. The percent ratios of EDI to ADI for acequinocyl,

spirodiclofen and carbendazim were 24.6, 22.7, and 58.5%, respectively. Results of EDI/ADI

exceeding 100% indicates a risk potential. Therefore, the results of this research indicate that the

detected pesticides are not harmful to humans. Although, the results show a negligible risk

associated with exposure via yuza tea consumption, a special precaution should be taken with the

possible total exposure to these chemicals from various foods in the future. Monitoring of

pesticide residue data in yuza has not been performed. Therefore, it is necessary to monitor the

residues of acequinocyl, spirodiclofen, and carbendazim in yuza continuously because of

possible health effects, widespread use, and insufficient residue data. Additionally, further

monitoring studies must be performed to improve food safety.

Pesticides levels in ginseng

The validated method was applied to the routine pesticide analysis of ginseng samples. Fresh

ginseng (n = 118), red ginseng (n = 24), and dried ginseng (n = 10) were analyzed following the

sample preparation method described above. Detailed data on the pesticide levels measured in

fresh ginseng are shown in Table 4. We detected 16 different pesticides in fresh ginseng samples.

Especially, tolclofos-methyl was detected in 102 samples (ranging from 18.25 to 404.5 μg/kg).

The detection level of fludioxonil was 56.62 μg/kg and detection frequency was 49.2%. The

residue level of chlorothalonil was higher than its MRLs, but detection frequency was not very

40

high at 0.9%. Detailed data of the residue pesticide detected in red ginseng are shown in Table 5.

The results show that the most frequently detected pesticide was tolclofos-methyl, which was

detected in 22 out of 24 samples at concentrations ranging from 13.14-119.4 μg/kg. The

detection frequencies of cyprodinil, fludioxonil, and difenoconazole were 79.0, 57.9, and 57.9%,

respectively, which were lower than their MRLs.

Table 6 shows residue levels detected in dried ginseng. The results indicate the existence of

quintozene, tolclofos-mehyl, cyprodinil, and difenoconazole. The detection frequencies of these

pesticides were relatively higher than those of others. The residue level of tolclofos-methyl was

higher than its MRL.

Kinetic parameters for pesticide degradation

Ten days after spraying, the degradation rates of chlorpyrifos and acequinocyl exceeded 90% and

100%, respectively. The degradation order of the seven pesticides was as follows: acequinocyl >

chlorpyrifos > spirodiclofen > carbendazim > deltamethrin > phosalone , prothiofos. Because the

half-lives of prothiofos, phosalone, and deltamethrin were longer than those of the others, their

application doses should be reduced. The doses of acequinocyl and chlorpyrifos could be

increased to improve productivity as they were under the MRLs. Three kinetic models were

employed to characterize the best-fit kinetic model. Among the theoretical models, FO and SO

models were the best-fit models for the pesticides residues, judging from the significance of the

coefficient of determination and the standard error. Therefore, it is recommended that the half-

life of the pesticide be assessed from the best-fit model rather than from the FO kinetic model.

CONCLUSION

In this study establishment of an analytical method to measure pesticides and to analyze pesticide

residue levels of Korean agricultural products such as yuza (Citrus junos Sieb. ex Tanaka), yuza

tea, and ginseng products was carried out. Risk assessments were also performed by calculating

estimated daily intake (EDI) and acceptable daily intake (ADI). In addition, kinetic study such as

the degradation order of pesticides in yuza tea was carried out. Acequinocyl, spirodiclofen and

carbendazim were detected in yuza samples in the concentration range of 0.07–0.15 µg/g, 0.11–

1.89 µg/g, and 0.03–5.15 µg /g, respectively. The percent ratios of EDI to ADI for acequinocyl,

spirodiclofen, and carbendazim were 24.6%, 22.7%, and 58.5%, respectively. The degradation

order of the seven pesticides in yuza tea was as follows: acequinocyl > chlorpyrifos >

spirodiclofen > carbendazim > deltamethrin > phosalone, prothiofos. For the accurate analysis of

Korean ginseng products a new analytical method was developed based on gas chromatography-

triple quadrupole tandem mass spectrometry (GC-MS/MS).

41

REFERENCES

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multiple actions. Biochemical Pharmacology, 58: 1685-1693.

Blasco, C., G. Font, and Y. Picó. 2002. Comparison of microextraction procedures to determine

pesticides in oranges by liquid chromatography–mass spectrometry. Journal of

Chromatography A, 970: 201-212.

Fujihara, S., and T. Shimizu, T. 2003. Growth inhibitory effect of peel extract from Citrus junos.

Journal of Plant Growth Regulation, 39: 223-233.

Goto, T., Y. Ito, H. Oka and H. Nakazawa. 2003. Simple and rapid determination of N-

methylcarbamate pesticides in citrus fruits by electrospray ionization tandem mass

spectrometry. Analytica Chimica Acta, 487: 201-209.

Jo, E., and K.G. Lee. 2012. Multiresidue pesticide analysis in Korean ginseng by gas

chromatography-triple quadrupole tandem mass spectrometry. Food Chem. 134: 2497-2503.

KCPA (Korea Crop Protection Association). 2006. Application for Pesticide use guide book.

Korea Crop Protection Association. <http://koreacpa.org/new/sub.html?sub=2>

Lan-Phi, N. T., T. Shimamura, H. Ukeda, and M. Sawamura. 2009. Chemical and aroma profiles

of yuza (Citrus junos) peel oils of different cultivars. Food chemistry, 115: 1042-1047.

Lee, K.G., S. Lee. 2012. Monitoring and Risk Assessment of Pesticide Residues in Yuza Fruits

(Citrus junos Sieb. ex Tanaka) and Yuza Tea Samples Produced in Korea. Food Chemistry

135: 2930-2933

Nam, Y., J. Hwang, and K.G. Lee. 2015. Pesticide residues in yuza (Citrus junos) cultivated using

ordinary and environmentally-friendly culture and their kinetic parameters for pesticide

degradation. J Pesticide Sci. 40: 60-64

Park, B. J., K. Son, and M.K. Hong. 2010. Monitoring of neonicotinoid pesticide residues in fruit

vegetable and human exposure assessment. Korean Journal of Pesticide Science, 14: 104-

109.

Taylor, M. J., K. Hunter, and S. Le Bouhellec. 2002. Multi-residue method for rapid screening

and confirmation of pesticides in crude extracts of fruits and vegetables using isocratic liquid

chromatography with electro spray tandem mass spectrometry. Journal of Chromatography

A, 982: 225-236.

Yoo, K. M., K. W. Lee, and I. K. Hwang. 2004. Variation in major antioxidants and total

antioxidant activity of yuza (Citrus junos Sieb ex Tanaka) during maturation and between

cultivars. Journal of Agricultural food chemistry, 52: 5907-5913.

Yamamoto, M., M. Nutahara, and H. Taniguchi. 1982. Residue of acaricides on fruits of yuzu

(Citrus junos Sieb. ex Tanaka) grown in vinyl-houses. Proceedings of the Association for

Plant Protection of Shikoku, 17: 29-34.

Kim, H. K., amd K. S. Lee. 2002. Effect of coverings on the Growth of ginseng and the

persistency of procymidone in growing soils. Korean Journal of Environmental Agriculture,

21: 24-30.

42

Walorczyk, S. 2007. Development of a multi-residue screening method for the determination of

pesticides in cereals and dry animal feed using gas chromatography-triple quadrupole

tandem mass spectrometry. Journal of Chromatography A, 1165:, 200-212.

43

Table 1. Validation parameters: Linearity, Limit of detection (LOD), Limit of quantification (LOQ), and Recoveries of pesticides used during yuza cultivation

Pesticides

Linearity LOD LOQ MRL

1

Recovery (%)

Precision2 (%)

Range

(μg/ml) Equation

Correlation

coefficient

(r2)

(μg/ml) (μg/ml) (μg/g) Intra-day

(n=5)

Inter-day

(n=5)

Chlorpyrifos 0.02-10 y=16.682x-0.9356 0.9992 0.02 0.06 0.5 109.9±3.9 10.0 9.4

Prothiofos 0.02-10 y=18.722x-2.7729 0.9972 0.03 0.10 2.0 99.4±4.2 10.3 5.9

Phosalone 0.02-10 y=18.116x-2.9475 0.9974 0.02 0.06 0.05 96.0±3.9 8.8 11.3

Deltamethrin 0.02-10 y=22180x-72217 0.9750 0.02 0.06 0.5 103.2±5.9 6.4 6.5

Acequinocyl 0.02-10 y=7773x-3737 0.9999 0.10 0.33 0.1/3.0a 93.8±6.9 15.9 6.9

Spirodiclofen 0.02-10 y=11818x-7.3200 0.9999 0.03 0.10 2.0 93.2±2.7 12.1 7.2

Carbendazim 0.02-10 y=42955x-560.10 0.9999 0.02 0.06 7.0 80.4±5.8 14.8 16.9

1Maximum Residue level

2 Relative standard deviation (RSD:%) of recovery rates from intra-day (n=5) and inter-day (n=5) experiments.

44

Table 2. Results of the pesticide residue analysis in Yuza cultivated by ordinary and environmentally-friendly culture

Detected

pesticide

Yuza cultivated by ordinary culture Yuza cultivated by environmentally-friendly

culture

No. of

samples

Analyzed

per year

Samples with

residues

Detection

range

(mg kg-1

)

No. of

samples

analyzed

per year

Samples with

residues

Detection

range

(mg kg-1

)

Korean

MRLs3)

(mg kg-1

)

2010 2011 2010 2011 2010 2011 2010 2011

Chlorpyrifos 50 1 (2%) 0 (0%) t 1)

-0.108 t 30 0 (0%) 0 (0%) ND

1) ND 0.50

Prothiofos 50 0 (0%) 0 (0%) ND2)

t 30 0 (0%) 0 (0%) ND ND 0.05

Phosalone 50 0 (0%) 0 (0%) t t 30 0 (0%) 0 (0%) ND t 2.00

Deltamethrin 50 0 (0%) 0 (0%) ND ND 30 0 (0%) 0 (0%) ND ND 0.50

Acequinocyl 50 0 (0%) 0 (0%) t t 30 0 (0%) 0 (0%) t 2)

t 1.00

Spirodiclofen 50 16

(32%) 5 (10%)

0.299

-1.889

0.274

-0.334 30

8

(27%) 0 (0%)

0.283

-0.801 t 2.00

Carbendazim 50 37

(74%)

27

(54%)

0.113

-5.148

0.088

-3.840 30

13

(43%)

8

(27%)

0.029

-2.907

0.111

-0.340 7.00

4)

1) t: LOD < values < LOQ

2) ND: values < LOD

3) MRLs: maximum residue limit of yuza tea

4) MRLs: maximum residue limit of other citrus fruits

45

Table 3. Exposure assessment parameters of pesticides in yuza tea samples

Pesticide

ALDa

(mg/kg)

EDIb

(mg/day) EDI/ADI

c100

Acequinocyl 0.4271 0.36541 24.6

Spirodiclofen 0.2040 0.17453 22.7

Carbendazim 1.1293 0.96592 58.5

a average level of detection. b estimated daily intake. c acceptable daily intake

46

Table 4. Pesticides levels detected in fresh ginseng (n=118)

Pesticides Mean(㎍/kg) Frequency (%)

Detection range(㎍/kg)

EBDC N.D. N.D. N.D.

Cadusafos N.D. N.D. N.D.

α-BHC N.D. N.D. N.D.

β-BHC N.D. N.D. N.D.

Quintozene 21.15 28.81 20.15-23.25

γ-BHC N.D. N.D. N.D.

Tefluthrin 7.34 0.85 7.34

Chlorothalonil 132.11 0.85 132.11

Tebupirimfos N.D. N.D. N.D.

δ-BHC N.D. N.D. N.D.

Tolclofos-methyl 34.62 86.44 18.25-404.52

Metalaxyl 17.62 40.68 10.25-36.62

Diethofencarb 27.62 4.00 26.25-28.62

Aldrin N.D. N.D. N.D.

Cyprodinil 16.26 24.58 15.62-24.25

Tolyfluanid N.D. N.D. N.D.

Flutolanil 38.63 35.59 26.92-65.62

Fludioxonil 56.62 49.15 7.69-56.29

p,p-DDE N.D. N.D. N.D.

Thifluzamid 20.63 6.78 7.20-47.29

Kresoxim-methyl N.D. N.D. N.D.

Dieldrin N.D. N.D. N.D.

Endrin N.D. N.D. N.D.

p,p-DDT N.D. N.D. N.D.

p,p-DDD N.D. N.D. N.D.

o,p-DDT N.D. N.D. N.D.

Trifloxystrobin 28.26 1.69 20.29-36.29

Carbosulfan 104.62 18.64 25.62-213.35

Fenhexamid 21.22 8.47 15.14-31.38

Cypermethrin 122.26 4.24 90.93-147.29

Difenoconazole 32.73 31.36 25.25-82.29

Azoxystrobin 18.36 3.39 4.29-34.17

47

Table 5. Pesticides levels detected in red ginseng (n=24)



EBDC N.D. N.D. N.D.




Quintozene 36.23 42.11 25.14-114.27


Tefluthrin N.D. N.D. N.D.

Chlorothalonil N.D. N.D. N.D.




Metalaxyl 16.15 47.37 13.15-23.35

Diethofencarb 34.17 15.79 32.15-37.98


Cyprodinil 29.25 78.95 19.38-115.18


Flutolanil 44.14 26.32 32.14-71.38

Fludioxonil 56.25 57.89 71.18-79.46


Thifluzamid 20.25 6.78 7.16-47.62







Trifloxystrobin N.D. N.D. N.D.

Carbosulfan 113.11 4.16 113.11

Fenhexamid 109.35 4.16 109.35

Cypermethrin N.D. N.D. N.D.

Difenoconazole 54.64 57.89 33.62-234.11

Azoxystrobin 26.24 4.16 26.24

48

Table 6. Pesticides levels detected in dried ginseng (n=10)



EBDC N.D. N.D. N.D.




Quintozene 25.14 50.00 25.14


Tefluthrin N.D. N.D. N.D.

Chlorothalonil N.D. N.D. N.D.




Metalaxyl N.D. N.D. N.D.

Diethofencarb 33.14 10.00 33.14


Cyprodinil 23.35 75.00 20.14-34.35


Flutolanil 33.35 10.00 33.35

Fludioxonil 71.41 37.50 71.13-71.73


Thifluzamid N.D. N.D. N.D.







Trifloxystrobin N.D. N.D. N.D.

Carbosulfan N.D. N.D. N.D.

Fenhexamid N.D. N.D. N.D.

Cypermethrin N.D. N.D. N.D.

Difenoconazole 37.32 50.00 35.26-39.52

Azoxystrobin 5.92 10.00 5.92

49

CURRENT SITUATION OF PESTICIDES USE IN

INDONESIAN AGRICULTURAL PRODUCTS

Agustina Asri Rahmianna1,Suharsono

2, and Didik Harnowo

3

1

Agronomist at Ecophysiology Division, Indonesia Legumes and Tuber Crops Research Institute,

Malang, East Java, Indonesia 2

Entomologist at Plant Protection Division, Indonesia Legumes and Tuber Crops Research

Institute, Malang, East Java, Indonesia 3 Director, Indonesia Legumes and Tuber Crops Research Institute,

Malang, East Java, Indonesia


ABSTRACT

The Green Revolution has remarkable contribution for food security for Indonesia and it is

highlighted by the achievement of rice self sufficiency in 1984. This success, however, generates

the new problem of over use of chemicals which emphasized on pesticides and fertilizers. In

regarding to economic, environmental and health issues, the Government of Indonesia has been

adopting Integrated Pest Management (IPM) strategy as the national policy for controlling pest

on agricultural crops since early 1986.The goals were economically and environmentally sound

along with the reduction of pesticide use. This policy has been supported by formal authorities

with law enforcements. The strong efforts undertaken by the Government of Indonesia and the

success of implementing IPM program were internationally recognized. The success of IPM in

rice was then adopted by vegetable and fruit farming as these crops are of high economic value,

and farmers need more pesticide and fertilizer to secure their high yield/production rather than

food safety and sound ecology. Despite of high achievement in IPM, the use of chemical

pesticides in some agriculture commodities is still threatening both economic and environment.

Keywords: Pesticide Residue, Misuse of Pesticide, Integrated Pest Management (IPM), Cost for

Pesticide

50

INTRODUCTION

Agriculture sector is responsible to feed people in the country. Increasing population will

increase food demand. Cereals, vegetables and fruits are staple foods and consumed every day by

almost all Indonesian, therefore, these are considered to be economically important. In order to

obtain high production of food crops, farmers or growers need packages of technologies that

generate high crop production. The technology should be economically profitable,

environmentally friendly, and technically acceptable. The package is composed of several

components of technologies i.e. appropriate cultural practices; high yielding variety; appropriate

weeds, pests and diseases control; optimum fertilizers; and water supply especially during the

critical periods of plant growth.

Regarding to plant protection, like growers in any agricultural country in the world,

Indonesian farmers rely on synthetic or chemical pesticides (mentioned as pesticide in the rest of

the paper) to obtain high yield of their crops. More than 95% farmers applied pesticides to their

crops to see instant effects (Gusfi, 2002 in Irfan, 2008). In the world, the biggest consumers for

pesticides are vegetables and fruits crops (26%), and consecutively followed by cereals (15%),

maize (12%), rice (10%), soybean (9.4%), cotton (8.6%), and the rest are other agricultural crops

(Dadang, 2007in Irfan, 2008). In regard to pesticide cost out of total production cost in Indonesia,

rice farming spent less pesticide than vegetable farming, with 0-10% compared to 26-50%,

respectively (Irfan, 2008). Due to the intensive use of pesticides, it is now considered to be over

use. And one possible reason came from the old paradigm saying the increasing yield or

production of agriculture commodities is a result of pesticide use. Since then, the Indonesian

farmers were attracted by the instantaneous effect of pesticides. As a result, phenomenon

showing increasing amount of pesticides used in Indonesia from 1986 to 1996, and from 17

thousand tons to 80 thousand tons, whereas in 2000 it reduced to 55 thousand tons. The

increasing demand on pesticides resulted in increasing money spent for importing pesticides

from 95,136 US dollars to 280,391 US dollars in 2006 and 2011, respectively. Meanwhile, the

number of registered pesticides was 813 in 2002 and hugely increased to 2,810 in 2013

(Republika online, 2014).

According to Crop Life Indonesia (the association of international agricultural companies),

the illegal and imitated pesticides are distributed in the markets with the value up to 10% of the

potential pesticides trading that reached 7 million – 8 million US dollars annually. As a

consequence the association has around 70 million US dollars loss annually (InfoPublik 2013).

One serious notion existing in the markets is that consumers want the “Cosmetic

Appearance” for all agricultural products, especially horticultural products. These high quality

products should have good appearance with no pest damage symptom. However, consumers

have not been intensively told about the essence of quality in term of nutrient content (nutritive

51

value) and pesticide residue. It is quite fair, therefore, to explore the effect of pesticides in that

beautiful appearance of horticultural produces, especially to human health. It has been very clear

that pesticides have a potent risk on human health. The pesticide can enter the human bodies

when they are exposed to pesticides at the time of carrying, storing and transferring them, and

mixing, spraying and cleaning spray equipments that have been used along the food supply chain.

This paper will discuss the current situation of pesticides in the farmers’ fields, integrated

pest management in Indonesia, and strategies to reduce the bad impact of pesticides through the

government regulations, and increasing the awareness on human health and ecological approach

with emphasis on biological as well as botanical pesticides.

CURRENT SITUATION OF PESTICIDES IN THE FARMERS’ FIELDS

Indonesian farmers have been spoiled by the presence of pesticides for many-many years. These

chemicals are easily found and freely sold in the markets. Despite the existence of government

regulation on the distribution of legal/permitted pesticides issued by Government Regulation No.

7/1973 on Supervision of Circulation, Storage and Usage of Pesticides (Statute Book of 1973 No.

12), the real condition in the fields shows that there are a lot of pesticides with high poisonous

content available in the markets. One report mentioned that in year 2013, there were 2,651

pesticides actively registered, and 183 pesticides did not actively register because of their unclear

detailed address of producers, expired permission letter, and others (Anonym 2013).

Pesticide and Human Risk

The ordinary farmers (farmers who stay in the village with minimum-medium literacy to

knowledge) have less understanding about pesticides. Instead as a toxic compound, farmers think

that pesticides are “medicine” for the crops to control the pest attack or fungal/bacterial diseases;

therefore, they are keen to buy pesticides even it is expensive. Farmers also tend to use pesticides

without paying attention to the dosage and instructions written in the package label determined

by the producers. As a result, many farmers apply higher up to much higher amounts than it is

recommended. Excessive amount of application results in high residue especially in vegetables.

Instead of the residual effect on vegetables, the excessive use of pesticides also increases the

chance of pesticides exposure on human body. The greater chance of exposure to pesticides may

increase the high incidence of chronic poisoning. Yuantari et al. (2015) reported that most

watermelon farmers in Grobogan District, Central Java had close contact of their body especially

their back, hands and feet as well as eyes and face when mixing and spraying pesticides.

Moreover, 40.7% of farmers used more than 10 types of active ingredients in a single mixing,

52

and 51.9% of farmers sprayed 6-10 tanks in one day. Another study conducted in Grobogan

District revealed that most farmers (>50% sampling farmers) had low level of knowledge on

those variables and the immediate improvement was suggested (Yuantari et al 2013). Also,

pesticide poisoned farmers who spray pesticide for their chili crops at Candi village, Bandungan

sub district, Semarang District in 2008 were tested if they have Cholinesterase in their blood,

and 26% or 74% farmers had moderate or mild toxicity, respectively. Due to little understanding

of the application procedures and the negative effects of pesticides, farmers apply abundantly to

their crops especially when there is pest outbreak. As a result there are high amount of chemicals

residues left on the leaves or fruits.

High incidence of pesticides illness among farmers occurs because:

Farmers are still lacking of knowledge (do not really understand) about the types of

pesticides as well as its active ingredients. Sometimes, farmers apply more than single

pesticide in one spraying by mixing those pesticides. They do not realize that mixing will

probably result in non effective effect. This practice often kills the non targeted pests.

It is very common that farmers apply pesticides not in the right time. For example:

application of pesticides in vegetables should be done 1-2 weeks before harvest. An

unwanted condition is even to spray one day before harvest.

In brief, risk factors related to pesticide toxicity relate to the variables of farmers knowledge,

attitude, types of pesticides, dosage of pesticide, frequency of pesticide spraying, wind direction,

personal hygiene,and personal protective equipment (Afriyanto, 2008).It is suggested that

farmers should use pesticides properly and wisely by reading the instructions on the package

label, spraying at a fix time and wearing personal protective equipment to maintain safety at

work (Yuantari et al., 2015).

Pesticides in Vegetables

Vegetables and fruits are the main source of food that is rich in vitamines, minerals, and essential

nutrients. Based on their nutrient status, vegetables and fruits are the main food that supports a

healthy life, and therefore they are considered to be high value economic crops. Other than

human consumption, these crops also can be used as aesthetics cropsto decorate the gardens.

Pesticides have been abundantly applied to horticulture crops especially leafy vegetables.

Other than excessive usage, farmers also had bad habits like cleaning the sprayers in the river

where the water is also used for daily life activities, and even more, they throw the pesticide pack

(sachets or bottles) away randomly in the farms.

Despite the negative effect of pesticides to human health, vegetable growers are still

applying a large amount of pesticides (especially insecticides) to their crops. In case of potatoes

crops grown in central production areas of Dieng High Land in Central Java Province, Potato

Grower Association has tried to campaign for reducing the use of pesticide and chemical fertilizer

since 2006. Only 170 farmers/growers (1%) join this program, and the remaining 99% farmers

are still intensively applying pesticides with huge amount as shown that 20% their production

cost is for chemical inputs (fertilizer and pesticide) in one cropping season (Pikiran Rakyat,

53

2014). One survey by Walhi Central Java reported that more than 90% of farmers had been

contaminated by pesticide from light-heavy contamination. The survey also indicated that

vegetable growers are more contaminated than rice farmers.Survey conducted by Regional

Health Office in Central Java found that from 217 sampling farmers; only 7% was free from

pesticides poison. Most farmers suffer from medium and light level of toxicity/poisonous, 55 and

35%, respectively (Sucahyo, 2014).

Residues of pesticides remained in food and environments have reached a dangerous level

for human health. The various levels and active ingredients of pesticide residue present in

vegetables such as cabbage, tomatoes, carrots and green leafy vegetables (Chinese cabbages,

Spinach), shallot, and chili (Ameriana et al., 2000; Munarso et al., 2006, Wariki et al., 2015). As

commercial commodities, these vegetables received high frequency of up to 10-15 times

pesticides spraying during crop growth period. It is very common that farmers are still spraying

few days before harvesting to keep the pests away for farmers to be able to get the very good

produce. It is not surprising, therefore, that human has me chemical compounds of pesticide in

the blood and breast milk (Frederik, 2012).

Horticulture farmers generally apply pesticides without paying any attention to the

recommended dosages. Most onion farmers (95.83%) located in Nganjuk District, East Java

Province who attended field school on IPM are still applying pesticides in the wrong dosage. It

means that only small number (4.17%) of onion farmers apply the right dosage. Almost all

farmers (98.96%) who have never attended the field school applied the wrong dosage

(Sulistiyono, 2002). Improper dosage builds the serious ecological impact. Applying dosage

different from its recommended in onion (Allium cepa) increased the main pest of Spodoptera sp

resistance to pesticide, as well as the population explosion of the secondary pest of Leromyza

Sp.(Sulistiyono,2002). This practice also destroyed beneficial predators such as Aranaeus inustus,

Argiope sp, Lycosa pseudoannulata, and Oxyopesjavanicus. This ecological disaster evenboosts

farmers to apply more pesticides, moreover inspires farmers to do self innovation to find the

suitable formulation to eradicate the pest or disease in their crops.

INTEGRATED PEST MANAGEMENT IN INDONESIA

Historical Review of Integrated Pest Management

Rice has been used as staple food for most of Indonesian people. Rice mainly uses as a source of

carbohydrate in the form of various processed foods such as steam rice, porridge, cookies,and

snack. Efforts to increase rice production have been initiated through Massive and Intensive

Guidance (it is called BIMAS and INMAS programs) in the early 1960s (Suharsono, 2001). In

this intensification program the Government of Indonesia (GOI) introduced five components of

54

technology named as “Panca Usaha Tani” – five principle components of technology for

producing rice i.e.inorganic fertilizers, high yielding variety, land preparation, irrigation systems,

and pest and diseases control. Fertilizers and pesticides were subsidized, capital investment was

facilitated by loan with low interest, and irrigation facilities were intensively developed in rice

production areas (Oka, 1995). Through this massive movement, harvest index and rice

production increased 3-4 times. During the National Rice Production Program from 1970’s to

1980’s, rice production grew with the rate of 3.74% annually (CBS 1973-1991 in Reksosudarmo,

2001).Accelerating high production of rice using intensification program brought Indonesia to

self sufficiency inrice by 1984 (Untung and Trisyono, 2010).

Despite the huge success of rice intensification program, the excessive use of pesticides

caused serious environmental problems. These include the incidence of pesticide poisoning on

human, cattle, poultry, and fishand other target species, agricultural products contamination,

reduction of natural enemies population, pesticide resistance of insect pests, more insect pest

outbreak, and chemical control not effective enough to kill pesticide resistant insect pests. To

correct them isuse and overuse of pesticides and reduce their impact on environment,

Government of Indonesia through Presidential Decree No. 3, 1986 launched a strategy of

integrated pest management (IPM) for rice in 1986.This program isa national policy, therefore,

all government agencies should support. The decree had the following objectives: (1). Improving

manpower skills, both farmers and agricultural officers at farm level, to implement the IPM, (2).

Increasing the efficiency of inputs use, particularly pesticides, and (3). Improving the quality of

environment and their effects on human health (Oka, 1995).The political support for IPM was

also obtained from the GOI by banning 57 pesticides for rice as written in the Presidential

Instruction No 3 year 1986 (Yanto, 2013).

Integrated Pest Management (IPM) is an effort to control pest population, level of plant

damage, and pest attack by integrating various control technique(s) to prevent economic loss and

environmental damages. Thus the IPM is optimizing the pest control using available control

techniques such as biological and natural control, cultural practices, and pesticides in economic

and ecological ways. Thus chemical control and other control techniques are also one of many

components of IPM. Through IPM strategy, the policy of pest control shifted from the method

that solely depended on pesticides to the one that used a combination of control tactics such as

synchronized planting, crop rotation, the use of natural enemies, and pesticides(Resosudarmo,

2001).

Along with the decree, the government has decreased subsidies for pesticides from 75-80%

of the total price in 1986 to 40-45% in 1987, and in January 1989 the subsidy was completely

eliminated. The Government also banned 57 broad-spectrum of pesticides (insecticides), and

allowed the use few relatively narrow-spectrum pesticides (Resosudarmo, 2001). In order to

activate the implementation of IPM program, the GOI established an Advisory Board in 1989.

The Board was responsible for the success of IPM program and ascertained the need of policy

improvement. By using “learning by doing” method, the field extension workers and pest

55

observers teach farmers. End of 1991, as many 2,000 extension workers and 1,000 pest observers

have trained 100,000 rice farmers, between 1991 to 1999 there were 200,000 farmers joined the

training each year. As many as 10% among them were received further training so that in turn

they trained rice farmers surrounding their IPM program. The cost of IPM training program in

1991-1992 was funded by the grant from USAID. Whilst in 1993-1998 the main source of

funding was the loan from World Bank. In 1999, the loan was terminated, and since then no

significant national IPM program has been implemented.

Success story of IPM on rice

In the class and field schooling, training of four pinciple philosophies of IPM was

introduced. These philosophies are techniques for producing healthy crops, conservation and

utilization of biologicals/natural control, good monitoring system of crop production, and farmer

as IPM expert. During the period of 1989-1999, field schooling for integrated pest management

has trained more than 1 million farmers who grow rice and other food crops. This achievement

was internationally acknowledged as a country that successfully implemented and developed

IPM (Yanto, 2013).

According to The Indonesian IPM Program Monitoring and Evaluation team, farmers who

joined the IPM program had their income increased by 50% in 1993. It was reported that farmers

who adopted IPM program reduced the use of pesticides by 56% and increased rice yield by 10%

(Oka, 1995 in Resosudarmo, 2001).

The success of IPM program was shown on controlling brown plant hopper (BPH)

Nilaparvata lugens Stal in rice that was adopted by rice farmers in almost all central production

areas in Indonesia (Untung and Trisyono, 2010). The following strategies were proposed to

reduce pesticide use to control BPH and other potential pests on rice:

1. Proper implementation of IPM as mentioned in Presidential Decree No. 3, 1986, Ministry

Agriculture Regulation No.887/Kpts/OT.210./9/1991, Government Act No. 12, 1992 and

Government Regulation No.6 Th. 1995

2. IPM program should be implemented in larger area (areawide approach)

3. The improvement of pest and disease detection and observation techniques

4. Review on the registration policy of newly pesticide for rice

5. Development new rice variety to BPH

6. Monitoring the new resistant biotype of BPH

As the IPM Program was launched, the knowledge in pest management of extension officer,

farmers and farmer groups was progressing through in-house training and field schooling. This

supports the Government Policy. The success of IPM in rice production has inspired the extent of

IPM program to different crops such as horticulture and estate crops. Despite IPM program has

56

been known to be successful in rice, the application of IPM other than rice crops, was initially

faced both technical and socio-economical constraints (Wardoyo, 1991 in Irfan, 2008). Later, the

success of IPM program on vegetable appears. For example, a study conducted in 2000 at 2

districts in West Java Province reported that cabbage growers who join the IPM training had

better knowledge, character and behavior on pesticide use than growers who did not join in IPM

program (Lubis,2001). On the other side, there is still the excessive use of pesticides for

vegetables eventhough the IPM program has been implemented (Munarso et al., 2006).

STRATEGIES TO REDUCE THE HAZARDOUS EFFECT OF PESTICIDES

The Government Policy

Since 50% of Indonesian people both directly and indirectly work in agriculture sector and the

daily menu of all Indonesian people are obtained from local products, the Government of

Indonesia is seriously responsible for the long-lasting life of the whole nation citizens. A series

of regulations start from the prioritized “the President Decree” and are enforced to be the

operational regulations “Minister’s Regulations”. These cover various issues from toxic and

dangerous residue materials management, plant protection, management on crop-disaster

organisms, and cultural-practice system. Moreover, the GOI is very serious about the pesticides

administration processes, starting from procedure and condition for pesticide registration, the list

of forbidden active materials or ingredients of pesticide, supervision of pesticide, the maximum

residue limit of pesticide, to supervision on the distribution, storage, and usage of pesticide.

The Government ActNo 12 year 1992 was stated in order to regulate the cultural practices

for food crops. It is mentioned that the application of pesticides should be in accordance to the 6

principles i.e. proper type, proper quality, proper target, proper dosage and concentration, proper

time, and proper application method and equipment. Further, it is emphasized that application of

pesticides should be as the last alternative method of pest control Law enforcement of IPM has

now under the umbrella of Ministry Agriculture Regulation No.887/Kpts/OT.210./9/1991 (crop

protection), Government Act No. 12, 1992 (production system) and Government Regulation

No.6 Th. 1995 (crop protection).

Increasing the Awareness of Farmers

A farmer’s behavior of using chemical pesticides was influenced by (1) farmer’s perception on

risk, the higher the risk perception the higher the quantity of chemical used; (2) farmer’s

perception on cultivar resistance to pests, the lower the tomato resistance to pest the higher the

quantity of pesticide used; (3) farmer’s knowledge on the danger of pesticide, the lower the

knowledge of farmers on pesticide danger, the higher the amount of pesticide used (Ameriana,

2008).

57

Generally, rice farmers are more rationale and would pay more attention on the negative

impacts of the use of pesticides when compared to vegetable farmers. Both farmers’ groups,

however, needed more attention and training to increase their knowledge on pesticides. The role

of local and central government is required to do extension work in these areas to deliver proper

pesticide use and application methods (Irfan, 2008). Farmers who never received any training on

IPM are still focusing their method of pest control by applying pesticide. This is because their

knowledge about pest ecology and non-chemical pest control are still limited (Prijono et al.,

2001 in Irfan, 2008).

Global/international markets where implement certain amount of maximum residue limits of

pesticides for food and beverages or any agricultural products should be positively and

enthusiastically responded by GOI together with farmers (Wiryadiputra, 2013). To enabling the

Indonesia agricultural products to compete in global markets, where consumers’major

requirement is pesticide residue free, therefore, farmers should be trained.

In increasing awareness of farmers to pesticide especially on factors related to pesticide

poisoning, avoid pesticide exposure, in the farmer’s activity. Donal and Paul (2014) suggest that

monitoring, counseling and guiding on how to correctly handle pesticide should be given

continuously by the competent authority. Whilst Maksuk (2014) suggest a recommendation or

risk management model for pesticide exposure in agriculture area.

Ecological Approach

Organic farming tries to minimize the negative impact to the surrounding environment, among

others by applying environmentally friendly pesticides. At present, the use of bio-pesticide and

botanical pesticide in the farmer’s level is not as familiar as synthetic pesticides. To be

successfully introduced to farmers, the economic assessment of this pesticide should be

undertaken.

Botanical pesticide is pesticide prepared from any part of plants i.e. leaves, fruits, roots,

tubers, flowers, and seeds. This sort of pesticide is relatively easy to prepare as the plant

materials are available, for example the goat weed (Ageratum conyzoides L.), nimb (Azadirachta

indica), and shallot (Alium sativum). The study revealed that application of goat-weed pesticide

in controlling brown plant hopper (BPH) Nilaparvata lugens Stal in rice, economically helps

farmers because it can reduce the application of synthetic pesticide that is recently more

expensive (Tabaet al.,2007). The extract of goat weed leaves is also effective in reducing rust

disease caused by Puccinia arachidis in groundnut leavesas well as substitutes the use of

chemical fungicide through the mechanism of delaying the germination of spores’ P. arachidis

and spores lyses as well (Yusnawan, 2013).

In the past 10 years, Indonesia Legumes and Tuber Crops Research Institute has developed

the ecological friendly pesticides for controlling pests and diseases in legumes and tuber crops. It

was found that numerous local plants materials either applied solely or combined with

58

entomopathogenic fungi can successfully reduce the pest population through the reduction in the

number of hatched eggs, resulting in reduced number of nymphs and adult pests. The good

example was the integration of botanical pesticide Annona squamosa seed powder or Jatropha

lenearis F + entomopathogenic fungi Lecanicillium lecanii; they can drastically reduce the

number of eggs of brown stink bug Riptortus linearis F. by 84% and 82%, respectively

(Suharsono and Prayogo, 2014).

CONCLUSION

The Government of Indonesia (GOI) has been seriously implementing the Integrated Pest

Management for rice and vegetable crops with various levels of success. The tropical condition

of Indonesia with high humidity and temperature, on the other hand, is a favorable condition for

the life and development of insect pests. This condition, therefore, is in need of using of

chemical pesticides as an agent for pest control. In other words, synthetic pesticides are still very

popular to farmers. The GOI through Ministry of Agriculture, therefore, is intensively managing

and strongly suggesting the wise use of chemical pesticides. Currently, the more friendly use of

pesticides has been introduced either by applying botanical or bio pesticides to substitute the

synthetic pesticides. However, farmers do not entirely rely on this ecological friendly pesticide in

managing their agriculture crops. The research on exploring local plants and microorganisms to

control pests and diseases is progressing and farmers with their own innovative feeling explore

the local materials to be available for controlling the existing pests.

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Candi village, Bandungan subdistrict, Semarang District. Post Graduate Thesis. 117 pp.

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vegetables free from pesticide residues (a case in tomatoes and cabbages). Jurnal

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Ameriana, M. 2008. Farmer’s behavior in using chemical pesticide on vegetables. Jurnal

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umum/13059-manajemen-pestisida-di-indonesia. Accessed 8 October 2015 (in Indonesian)

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terdaftar-harus-dikurangi. Accessed 10 October 2015.

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memprihatinkan/2440832.html. Accessed 06 October 2015. (in Indonesian).

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Suharsono, 2001. Resistance assessment of some soybean genotypes to pod sucker Riptortus

linearis F. (Hem.: Alydidae). Doctor of Philosophy Dissertation. Gadajahmada University.

163 hlm. (unpublished) (in Indonesian).

Suharsono, and Y. Prayogo. 2014. Integration of botanical pesticide and entomopathogenic fungi

to control the brown stink bug Riptortus linearis F. (Hemiptera: Alydidae) in soybean.

Journal of Tropical Plant Pests and Diseases14(1): 41-50.

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case in District of Nganjuk, East Java). Post Graduate Thesis. Bogor Agriculture Institute.

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Suwahyono U. 2009. Procedure and Guideline for bio-pesticide making.Penebar Swadaya.

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Indonesian)

Taba, A.H., Y. Surung, and I.N.R. Parawansa. 2007. The evaluation on extension and farming

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stink bug Nilaparvata lugens stal. Jurnal Agrisistem Socio-economical and extension Series.

3(2): 95-101.

Untung, K. and Y. A. Trisyono. 2010. The threat of brown stink bug to rice self sufficiency.

Executive Summary. 9 p. (in Indonesian).

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prophenophos residues on tomato plants in Langowan West Subdistrict North Sulawesi.

Jurnal Ilmiah Sains. 15(1): 47-51. ejournal.unsrat.ac.id (in Indonesian).

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the efforts to solve the problem. Review Penelitian Kopi dan Kakao 1(1): 39-61 (in

Indonesian).

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of pesticides (A study conducted at Curut Village, Penawangan Subdistrict, Grobogan

District). Proceeding of National Seminar on Natural and Environmental Management. 2013.

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Yuantari, M.G.C., B. Widianarko, and H.R. Sunoko. 2015.Pesticide exposure risk analysis

against farmer’s health. Jurnal Kesehatan Masyarakat 10(2): 239-245 (in Indonesian)

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to control peanut rust disease and phytochemical screenings of secondary metabolites.

Journal of Tropical Plant Pests and Diseases. 13(2): 159-166.

61

CURRENT USE OF PESTICIDES IN THE AGRICULTURAL

PRODUCTS OF CAMBODIA

Visarto Preap1

and Kang Sareth2

1,2Department of Plant Protection Sanitary and Phytosanitary,

General Directorate of Agriculture (GDA),

Ministry of Agriculture Forestry and Fisheries (MAFF),

Kingdom of Cambodia

e-mail: [email protected]

ABSTRACT

Pesticide Management is still a complicated task with cross mandatory responsibility from

various departments. At the Ministry of Agriculture Forestry and Fisheries (MAFF)in Cambodia,

there are three departments working namely: 1) the Department of Agriculture Legislation as

regulatory authority in charge of pesticide registration, licensing, inspection;2) PPSPS

Department of the General Directorate of Agriculture (GDA), playing a role as technical adviser

in field evaluation of pesticides and efficacy field testing for registration; and 3) the National

Agricultural Laboratory of GDA, which has a role in the analysis pesticides. Lack of law

enforcement to farmers especially those who greatly depend on using pesticides created a big

challenge for pesticide management in Cambodia; banned and restricted pesticides are widely

available in the local markets. Counterfeit and illegal pesticide products are often found in

unregistered pesticide shops/retailers. To fight against these challenges Cambodia shall take

priority actions: improve border inspection on circulation of pesticides, and strengthening

pesticide management law.

Keywords: Pesticide law enforcement, counterfeit pesticides, banned products, restricted, and

permitted pesticides, border control, quality testing and residue analysis.

62

INTRODUCTION

Cambodia is located at 102o to 108

oE and 10

o to 15

oN. This country is influenced by the

Monsoon climate, which consists of two seasons: dry season (November – April) and wet season

(May – October). The rainfall is varied from 1250 to 2500 mm annually; the lowest is in January

and the highest is in October with humidity range from 69% to 80%, the lowest of which occurs

in March and the highest is in September. The day length is from 11 hours to 13 hours. The

shortest day length is in December and longest is in June. The temperature varies from 23 to

33oC, the lowest occurs in December and highest is in April; with an evaporation of 2230mm per

year. The lowest and highest evaporation occurs between September and March. The natural

resources of Cambodia are officially under the protection of the State, as expressed in Article 59

of the constitution: “The state shall protect the environment and balance of abundant natural

resources and establish a precise plan of management of land, water, air, wind, geology, ecology,

ecologic system, mines, energy, petrol and gas, rocks and sand, germs, forests and forest

products, wildlife, fish and aquatic resources”. The land use is divided by main groups such as

Agricultural land (24%), forest cover (56%), grassland (6%), shrub land 10%), soil-Rock (0.2%),

urban (0.1%) and water (3%).

Three departments under Cambodia’s Ministry of Agriculture, Forestry and Fisheries

(MAFF) are responsible for plant protection and pesticide management: (1) the Department of

Agriculture Legislation (DAL) as regulatory authority in charge of pesticide registration,

licensing, and inspection, 2) Department of Plant Protection Sanitary and Phytosanitary

(DPPSPS) under the General Directorate of Agriculture (GDA), playing a role as technical

adviser, provide training on pest management, pest control technology, pest monitoring, pest

forecasting, pest outbreak warning, invasive species control and general pesticide advisory. It

also conducts researches and developments on various pests to strengthen the implementation,

managing pesticides including pesticide registration (providing efficacy field testing), and

development of recommendation on pesticide use, and (3) the National Agricultural Laboratory

(DAL) of the General Directorate of Agriculture, which has a role in the analysis of pesticides

63

Pesticide use:

Cambodia is rich in bio-diversity and accordingly, agricultural crops are diversified. With the

introduction of modern agriculture in the early `60s, traditional crop varieties were replaced by

modern crop varieties (MCVs). These high inputs which are responsive to MCVs brought a

significant change in Cambodia’s agriculture. In due course, pest dynamics has also changed and

a number of pest outbreaks occurred frequently. To overcome these problems, use of chemical

pesticides also became frequent.

Cambodia’s agriculture policy has emphasized eco-friendly production system, organic

farming and IPM practice for sustainable agricultural development and food safety. Considering

all these issues, the Pesticide Registration and Management Department has also emphasized the

registration of bio-pesticide pesticides, which gradually reduce highly hazardous pesticides.

The Plan and policies of Cambodia also encouraged eco-friendly measures of agricultural

production, IPM practice and organic farming which directly or indirectly support the concept of

pesticide risk reduction in food safety. The preparation of pesticide policy and bio pesticide

promotion directives is under way which encourages for the production, registration and use of

bio-pesticides pesticides and bio-agent. The Royal Government of Cambodia (RGC) is regularly

organizing training and awareness program on the safe use of pesticides to stakeholders and

users of the pesticides.

Cambodia has also signed and ratified the Stockholm Convention (POP), Montreal Protocol

(Ozone Depletion Materials) and Basel conventions with full developed action plans for

implementation of the first two conventions with focal points placed in the Ministry of

Environment. Whereas MAFF / DAL is the focal point for the Rotterdam Convention which had

been acceded by Cambodia since May2013.

Cambodia’s pesticide market has continued to expand over the last decade, which is basically

a result of the liberalization of Cambodia’s economy. Cambodia has no pesticide manufacturing

capacity of its own, and most available pesticides are imported officially and illegally from

neighboring countries such as Thailand and Vietnam. Some of the most popular pesticides, such

as the organophosphates methyl parathion and Mevinphos are extremely hazardous and are

banned according to Cambodian law. The Environmental Justice Foundation (EJF) reported that

inappropriate pesticide use in agriculture is widespread and that products are used by untrained

and often illiterate farmers, who incur serious health consequences.

In recent years, MAFF has made strong efforts in pesticide management in Cambodia. The

Government has issued an order to all relevant units to strengthen pesticide management and

quality control including across border trade, distribution, sale and use of agrochemicals in the

country. The Government has enforced pesticide labeling regulations, including development of

labels in local Khmer language in line with the FAO Code of Conduct on the Distribution and

the Use of Pesticides. In addition, MAFF is reviewing and updating the pesticide list including

banned, restricted and permitted products through Ministerial proclamation No. 484 MAFF dated

26 November 2012 (Table 1).

64

Tabel1- List of Banned/Deregistered Pesticides in CambodiaS.N Name of pesticides Year of banned S.N Name of pesticides Year of banned

1 Azinphos Methyl 2012 50 Demeton-s 2012

2 Aldicarb 2012 51 Demeton-S-methyl 2012

3 Aldoxycarb 2012 52 Diamidafos 2012

4 Aldrin 2012 53 Dichlorophene/Antiphen 2012

5 Aminocarb 2012 54 Dieldrin 2012

6 Amitraz 2012 55 Difenacoum 2012

7 Amitrol 2012 56 Difenthialone 2012

8 Antu 2012 57 Dimefox 2012

9 Aramite 2012 58 Dimethilan 2012

10 Arsenic compound 2012 59 Dinoseb 2012

11 Beta-HCH 2012 60 Dinoterb 2012

12 Benomyl 2012 61 Dioxathion 2012

13 Binapacryl 2012 62 Edifenphose 2012

14 Bromethalin 2012 63 Elemental phosphorous 2012

15 Bromophos 2012 64 Endosulfan 2012

16 Butoxycarboxim 2012 65 Endothion 2012

17 Cadmium compound 2012 66 Endrin/ Nendrin 2012

18 Cadusafos 2012 67 EPN 2012

19 Calcium arsenate 2012 68 Ethoprop(Ethoprophos) 2012

20 Calcium cyanide 2012 69 Ethylene dichloride(EDC) 2012

21 Camphechlor 2012 70 Ethylene oxide 2012

22 Captafol 2012 71 Famphur(Famophos) 2012

23 Captan 2012 72 Fenamiphos 2012

24 Carbon tetrachloride 2012 73 Fenbutatin oxide 2012

25 Carbophenothion 2012 74 Fensulfothion 2012

26 Chlordane 2012 75 Fentin hydroxide 2012

27 Chlordecone 2012 76 Fluoroacetamide 2012

28 Chlordimeform 2102 77 Fonofos 2012

29 Chlorethoxyfos 2012 78 Fosthietan 2012

30 Chlorfenvinphos 2012 79 Furathiocarb 2012

31 Chlorophenols 2012 80 HCH(Insecticide) 2012

32 Chlormephos 2012 81 Heptachlor 2012

33 Chlorbenzilate 2012 82 Hexachlorobenzene(HCB) 2012

34 Chlorophacinone 2012 83 Isobenzan 2012

35 Chlorthiophos 2012 84 Isodrin(Isomer of Aldrin) 2012

36 Copper arsenate 2012 85 Isoxathion 2012

37 Coumaphos 2012 86 Leptophos 2012

38 Crimidine 2012 87 Lindane(Gamma-HCH) 2012

39 Crotoxyphos 2012 88 Medinoterb acetate 2012

40 Cupric acetoarsenite 2012 89 Mephospholan 2012

41 Cyanthoate 2012 90 Mercaptofostion 2012

42 Cycloheximide 2012 91 Methiocarb 2012

43 Cyhexatin 2012 92 Methomyl 2012

44 Daminozide 2012 93 Mevinphos 2012

45 DBCP(Dibromochloropane) 2012 94 Mexacarbate 2012

46 DDT 2012 95 Methamidophos 2012

47 Demephion-o 2012 96 Monocrotophos 2012

48 Demephion-s 2012 97 Oxamyl 2012

49 Demeton-o 2012 98 Parathion-methyl 2012

In the context of chemical management, the Ministry of Environment is starting to introduce

the GHS (Global Harmonization System) for pesticide labeling. Japan International Cooperation

Agency (JICA) has provided assistance for staff capacity building and facilities for strengthening

pesticide analysis laboratory. Frequent inspecting to the pesticides retailers, dealers, formulators

and others users are done in order to check whether they follow the code of conduct related to

pesticide use as regulated by the RGC. Pesticide laboratory has been established and brought into

operation especially on formulation and quality testing. With regards to pesticide management,

there are many challenges which remain unresolved, such as insufficient enforcement of rules

and regulations, uncontrolled importation, and broad availability of undesirable pesticides,

misuse and over use, limited data on health and environmental effects and high pesticide residues

in food. However, on a positive note, there has been a broad recognition throughout the

Government, NGOs and private sector with regards to current pesticide issues and their negative

65

implications for production, health, environment and trade. However, testing for residues in

fruits and vegetables is the first priority for the upcoming action plan.

Through Policy Component of project GCP/RAS/229/SWE of the Swedish-supported

Pesticide Risk Reduction Program activities were carried out to address the issue of highly

hazardous pesticides through capacity building for chemicals management in general. Cambodia

focused on initial steps to develop adequate regulatory framework including legal documents and

functional mechanism for the control of pesticides and capacity building for staff members in

national and provincial levels on pesticide management including training for retailers and

inspectors, pesticide quality control and inspection, and registration and data base development.

In addition, Cambodia participated in an FAO Regional High-Level Workshop on Licensing and

Inspection of Pesticide Sellers, which was organized in Hanoi on 10-11 November 2008. The

workshop facilitated the exchange of information and experiences, and to discuss common

issues/questions related to the establishment and operation of a functional licensing and

inspection schemes for pesticide importers and sellers in compliance with legal obligations.

Farmers and pesticide traders’ survey took part to determine the perceptions and practices of

pesticide use among farmers that incorporated agrochemicals into their farming strategy. Survey

results in nine provinces, identified to be potentially at elevated risk from pesticide use, were

surveyed. In Cambodia, specialist traders are found in larger towns that sell pesticides, together

with seeds and chemical fertilizers. In smaller towns general stores sell pesticides typically

alongside other products, including groceries and cosmetics. Of the 109 pesticide retailers

interviewed, 10 were owners of specialized stores and 99 were general store owners. Farmers are

more willing to trust the quality of pesticides purchased from specialized stores. It is often

suspected that general stores dilute their stocks and due to low turnover, some pesticides turn out

to be older stocks. However, sometimes distance prohibits farmers from travelling to major

towns to buy pesticides from specialized stores. In some cases, farmers do not have enough

money to buy pesticides and local stores will often sell on credit, whereas specialized stores will

not. Sometimes local stores will sell pre-mixed ‘pesticide cocktails’. The survey found that 97%

of traders were selling more pesticides and were giving them more shelf space. Pesticide traders

said they had no difficulty in acquiring pesticides and assumed that all pesticides were legal in

Cambodia. All pesticide retailers were unaware of the 1998 sub-decree on Standards and

Management of Agricultural Materials that lists pesticides that are banned or of restricted use in

Cambodia.

The chemical products used in Cambodian agriculture are mainly fertilizers and pesticides.

While Cambodia is not an agro-chemical producer (table2); and mainly imports from

neighboring countries (Thailand & Vietnam), there are some cases in which pesticides are

imported from China and European Union members states.

According to the law, agrochemical importing companies must be registered at the MAFF

before allowing them to be imported into Cambodia. However, there were not all agrochemical

importing companies which have been registered; some of them still imported without

registering their products. Some unofficial reports indicated that around60-80% of imported

pesticides were illegally imported bought along the Thai and Vietnam borders. Official data

recorded that till now December 2013 registered pesticides are used in Cambodia are 750

66

Bamate Monitor 50SC

Bent 600 Monitor 60SC

Filitox 50EC Monitor 70DD

Filitox 60DD Methon

Filitox 70SC Morris

Giant 50%WW EC Ovansu

Giant 70DD Sigtifos

L. Talon Siter-Nissan

Marathon Tom 50EC

Methamidophos 70SL Tom 60EC

Methamidophos 60 Tom 70EC

Methamidophos 70% U-T 70

Methaphos 70SC U-T 80

Monitor 50EC Vindo

Table2- Pesticide data in brief

No. Description Number

1 Pesticide registered from 01 Oct 2012 to 10th

Dec 2013 750

2 Pesticide registered till 01Oct 2012 to 10th

Dec 2013 750

3 License holder (Retailers) 35

4 Trained personnel (Safe storage and Use) 9300

common names and trade name of pesticides, while the retailers holding license, which directly

deliver to the users are only 35 holders (Table 2).

Pesticide use: Before1980, farmers who used pesticides in their farm production were only

7%. The number of pesticide consumption increased up to 49% in 1985-1994 in donation

framework (Anonymous, 2009). A basic study in 2004 reported that about 67% of farmers used

chemical pesticides in their production crops at least one time a year. The use of chemical

pesticides also varied depending on the seasonal crops; high usage of pesticides occurs during

the dry season crop, where 98% of the community use chemical pesticides in their vegetable,

tobacco, bean and dry season rice crop (8-15% of the wet season rice crop) (Anonymous, 2009).

There is no records of the exact volume of chemical pesticides used; but it were estimated about

3,570 tons were used in 2007.

Fig. 2. The 28 trade names of Methamidophos

Type of pesticide use: there are 522 trade names of 133 common names of chemical

pesticides available in local markets; most of them are unregistered pesticides. We found several

trade names in one common name of those unregistered pesticide such as Methamidophos and

Abamictin (Fig. 2 & Fig. 3). Unregistered/illegal pesticides mostly are extremely and highly

hazardous pesticides (Fig. 1). There were 13 common names of banned and restricted pesticides

for use observed in the markets in 2007; they are Methyl parathion, Mevinphos, Methamidiphos,

Methomyl, Monocrotophos, Dichlophos, DDT and Chlodane.

67

Ababest kabamax

Abalimec 36EC Masket

Abamectin Masta

Abamet Maxagro

Abatimec 1.8EC Naicer

Abatin 1.8EC Pivbek

Amerec 36EC Plutel 3.6EC

Avina Pro 3K

Bambin Saco-sdim

Citrameth-Luxen Samsin

Fanty 2.5 EC Sitramec

Geno mectin Ste Kodsilla

Hen Abid Supermectin

Intak Voi Thai 2.6EC

Inter Face Vertimec 1.8 EC

Jacket World mekin 2EC

43%

32%

9%

16%

0%

5%

10%

15%

20%

25%

30%

35%

40%

45%

50%

1a II 1b others

1a

II

1b

others

Fig. 3. Some trade names of Abamictin

Several studies on pesticide impact showed that Cambodian farmers used a highly hazardous

pesticide which has been well recorded. FAOs’ study indicated that the percentage of using

highly hazardous pesticides class ‘1a’ decreased from 77% to 43% in 2008. Moreover, the study

found that: among the 84% of farmers using highly hazardous, there are 43% using ‘1a’ class,

9% using ‘1b’ class and 32% using II class (Fig. 1).

Fig. 1. Percentage of pesticide used in different class of WHO’s classification

(Anonymous, 2009)

68

Cambodian farmers commonly use at least 12 insecticides belonging to the neonicotinoid

family (Cheang, 2013). These insecticides are not yet included in the country’s updated list of

banned or restricted pesticides (MAFF 2012).

Indiscriminate use of pesticides not only puts sustainable agricultural production at risk

through the disruption of vital ecosystem services, pesticide residues on fresh produce that

exceed the maximum(allowable) residue limits (MRLs) also raise food safety concerns and

jeopardize their export potentials. MRLs are standards set by individual countries for traded

agricultural commodities according to types of pesticides. Pesticide residues result from: 1)

heavy pesticide use on the growing crop;2) insecticide used in post-harvest management to

preserve food during storage; and 3) the persistence and carry-over effect of residues in the soil.

Survey studies of pesticide contamination of vegetables in Cambodian markets found

produce containing residues of organochlorine (Wang et al., 2011), organophosphate and

carbamate (Neufeld et al., 2010) exceeding MRLs. Cambodia ranks first among 13 countries in

the region with the highest pesticide residue on vegetables, particularly leafy vegetables from

Kandal province (Wang et al., 2011).

Based on pesticides displayed in the markets for sale, the study found that the majority of

pesticides being used in Cambodia were in ‘1a’ class of WHOs’ classification with the 18% of

common names and 2% of trade names. Moreover, the POPs’ pesticides such as DDT and

Chlodane were still found to be used in the farms (EJF, 2002).

Even with the National targets and efforts regarding pesticide risk reduction as well as

phasing out highly hazardous pesticides, Cambodia is still facing a big obstacle due to the

limitation of human resources for anti-counterfeiting/illegal pesticides and farmers still prefer to

use pesticides because their perception on pesticide use is that it is the more the effective option

(Anonymous, 2009).

Several unregistered shops have signages in foreigner languages (Thai & Vietnam), carrying

those pesticides which are banned for use in Cambodia and are not available in those countries.

Moreover, the labels are too old and already expired (EJF., 2002).

Pesticides are displayed for sale together with other

food stuff in unregistered shops.

Pesticide application service provider is available

at the farm level.

69

Pesticides disposal: However, the Ministry of Agriculture, Forestry and Fisheries (MAFF)

has taken the priority action to minimize this hot issue. The sub-decree no. 69 was not fully

implemented in the pesticide management departments; therefore the Law on the Management of

Pesticides and Fertilizers has been ratified by the National Assembly on December 21st, 2011.

Future actions:

As chemical pesticides are widely available almost practically anywhere and most of them are

illegal, counterfeit products in Cambodia should take priority action as suggested below:

- Improve pesticide movement inspection activities in the borders, which is a key part in

strengthening pesticide management;

- The limitation of law enforcement with lack of public awareness should have the following

priority actions: Strengthen Law enforcement especially on Law on the Management of

Pesticides and Fertilizers, The government should lead the push for the enforcement of

policies, regulations and legislation relating to environmental protection and the responsible

use of pesticides to reduce the risks and impacts of agro-chemicals use. The implementation

of the law should be mandatory, not voluntary. Financial or other penalties should be

imposed on traders for illegally importing, selling or distributing banned pesticides, and on

farmers for using them. Information about the law should be well disseminated and explained

to farmers, retailers, importers and border inspectors through newspapers, television, radio

and other forms of media.

- Focus on reviewing and revising pesticide management legislation toward reducing highly

toxic pesticides (Ministerial declaration on banned, restricted, and permitted pesticides for

use);

- Step-by-step phase out of highly toxic pesticides; use pesticides in accordance with

regulations related to ensure safety for humans, animals, plants, environment and food;

- Promote study and application of science and technology to production, trading and use of

bio-pesticides and other environmentally friendly control measures; develop pest-free areas.

Educational efforts, especially at the farm level, are needed to improve compliance with

ASEAN GAP standards. Furthermore, the global and domestic markets for organic foods and

beverages are growing and the demand for food safety is increasing.

- Build a system for waste container collection and treatment; use containers made from

recyclable materials;

- Encourage traders and plant protection service organizations to provide training and guiding

for safe and effective use of pesticides; provide technical training for manufacturers;

- Encourage IPM and GAP measures; encourage plant protection service activities, organize

specialized agricultural technical services.

70

CONCLUSION

The Ministry of Agriculture, Forestry and Fisheries (MAFF) in Cambodia is playing an

important role to solve the sensitive issue of the national socio-economic, environment and

agriculture sector; this ministry should lead the enforcement of policies, regulations and

legislation related to environmental protection and the responsible use of pesticides to reduce the

risks and impacts of agro-chemicals use. In MAFF there are three departments responsible for

the regulatory authority in charge of pesticide registration, licensing, inspection (DAL), technical

adviser infield evaluation of pesticides and efficacy field testing for registration (PPSPSD), and

analysis of pesticides for supporting counterfeit and illegal pesticides policy (NAL).

The problem with use of pesticides takes its roots based on the limited knowledge of

pesticide use along with lack of law enforcement. This has created a big challenge for pesticide

management in Cambodia. Most banned and restricted pesticides are still available in the local

markets. Cambodia shall take the prime actions to improve border inspection on the movement

of pesticides; this action is part of firing against counterfeit and illegal pesticide products; which

is very often found in unregistered pesticide shops/retailers.

REFERENCES

Anonymous. 2009. in Khmer language ‘Report on Chemical Management in Cambodia’

CEDAC. 2004. Pesticides use and consequence in Cambodia. Centre d’Education et de

Development Agricole Cambodgien.

EJF. 2002. Death in Small Doses: Cambodia’s Pesticides Problems and solutions. Environmental

Justice Foundation, London, UK.

Cheang H. (2013), Pesticides in Cambodia (PhnomPenh: Royal University of Agriculture)

Neufeld, D.S.G., H. Savoeun, C. Phoeurk, A. Glick and C. Hernandez (2010), “Prevalence and

Persistence of Organophosphate and Carbamate Pesticides in Cambodian Market

Vegetables”, Asian Journal of Water, Environment and Pollution, 7(4): 89–98

Ministry of Agriculture, Forestry and Fisheries (2012),“List of Banned Pesticides and Restricted

Pesticides”,Prakas No. 484

Wang, H.S., S. Sthiannopkao, J. Du, Z.J. Chen, K.W. Kim, M.S. Mohamed Yasin, J.H. Hashim,

C.K. Wongand M.H. Wong (2011), “Daily Intake and Human Risk Assessment of

Organochlorine Pesticides (OCPs) based on Cambodian Market Basket Data”, Journal of

Hazardous Materials, 192(3): 1441–1449

71

PESTICIDE RESIDUES IN FOOD AND THE ENVIRONMENT IN THE

PHILIPPINES: RISK ASSESSMENT AND MANAGEMENT

Cristina M. Bajet

National Crop Protection Center, Crop Protection Cluster

University of the Philippines at Los Banos

College, Laguna 4031, Philippines

email: [email protected], [email protected]

ABSTRACT

The Philippines as a developing country and with limited resources to implement intense monitoring of pesticide residues has focused on other ways to assess and manage risks. The top priority of the Philippines is food production/security and in the local market, pesticide residues analysis as a basis for market acceptance is secondary. Compliance to Maximum Residue Limits (MRLs) of major export crops like banana, pineapple, mango, okra, and asparagus, among others is the main focus of the Government. Rice is the largest consumer of pesticides in terms of volume but the pesticide application in high value crops is more intense.

The paper is a review of Philippines’ efforts on pesticide residue related problems. Focus is on pesticide regulation, monitoring and research activities of the Government Laboratories doing pesticide residue analysis on food and/or raw agricultural commodities. Rapid detection tools like colorimetric test kit and enzyme inhibition tests as solution for monitoring residues of organophosphates and carbamates on selected vegetables is the focus of the paper. Discussion includes the results of research on pesticide residues in water and soil and other environmental concerns related to pesticide usage in agriculture, to include ecotoxicological studies on non-target aquatic organisms.

Management options include regulatory control and promotion of organic agriculture through the enactment of the organic act. The food safety law puts together options for safe food available locally and export products by minimizing trade risks due to pesticide residues through generation of locally generated data.

Keywords: rapid test kit, regulatory control, organophosphate and carbamate pesticides, dietary risk, environmental risk



72

INTRODUCTION

The Philippines effort is to monitor various farming activities from conventional (to include

traditional, Integrated Pest Management (IPM), Good Agricultural Practices (GAPs), etc.) to

organic farming and with limited resources. This situation necessitates a concerted effort by

Government in partnership with stakeholders for pesticide residue monitoring and evaluation to

ensure consumer protection and food safety. With limited resources to implement intense

monitoring of pesticide residues, the Philippines have focused on other ways to assess and

manage risks. The top Government priority is food security. Food safety like pesticide residues

analysis as a basis for market acceptance is secondary. Compliance to Maximum Residue Limits

(MRLs) of major export crops like banana, pineapple, mango, okra, and asparagus, among others

is the focus and priority of the Government. Rice is the largest consumer of pesticides in terms of

volume but pesticide application in high value crops are more intense, resulting to residues on

the crops. Residues on rice resulting from pre harvest are minimal and most residues in rice

maybe related to post harvest application. For high value vegetables available in the market, the

intensity of pesticide application and the concentration of residues is correlated to the value of

the crop, production season and/or consumer demand. During festivities and the Christmas

season, consumer demand increases and the farmers are pressured to protect their crops because

the value is higher, resulting to higher pesticide residues.

Regulatory control

Pesticide residue risk assessment on food safety and management of pesticide use is

regulated through pesticide registration. The government agency mandated to regulate

availability and use of pesticides is the Fertilizer and Pesticide Authority (FPA) created with

Presidential Decree 1144, now under the Office of the President. All pesticide products are

evaluated by a team of experts on pesticide specification, bio-efficacy, toxicology, residues,

environmental effects and environmental fate and transport data. In cooperation with FPA, the

Bureau of Plant Industry National Pesticide Analytical Laboratory (BPI-NPAL) and its satellite

Pesticide Analytical Laboratories (PALs) provide laboratory services for formulation analysis as

part of the registration requirements to check the quality of pesticide products for registration. As

part of the food safety assessment and regulation and as a requirement for registration, or

approval for label expansion to other crops, a local residue data is required and a dietary risk

assessment is done based on Filipino diet.

Likewise, BPI NPAL and its five satellite PALs nationwide do residue analysis as part of

market basket monitoring of residues. Results of these monitoring activities are also a basis of

FPA regulation. Pre export analysis is also done by BPI-NPAL mainly for mango and okra

while crops from corporate growers like Dole, Del Monte, Stanfilco, among others, are

responsible for their exports in terms of pesticide residues.

Part of FPA’s regulation is training of stakeholders of the pesticide industry. Researchers

involved in bio-efficacy and supervised residue trials are to be licensed. Pesticide Industry

personnel undergo training and licensure as accredited responsible care officers and are actively

73

involved in the partnership with FPA on the Government’s Stewardship Program. Pesticide

Applicators and Pesticide Marketing Personnel are also certified and licensed by FPA.

The setting of Maximum Residue Limits (MRLs) is part of the FPAs regulatory process but

the Bureau of Agriculture and Fisheries Standards (BAFS) is the Government agency that is in

charge to convene all experts, stakeholders and relevant Government Agencies to set the national

MRL based on the existing data and pesticide registered usage as approved by FPA. Regulatory

control also include the passing of laws related to food safety Republic Act (RA) 10611 and to

organic agriculture act of 2010 RA 10068, which is one of the main focus of the current

Secretary of the Department of Agriculture (DA).

Monitoring of residues using rapid detection kits

In support of Government efforts to minimize risk due to the use of pesticide products, research

on pesticide residues in food and the environment is part of the research activities at the National

Crop Protection Center (NCPC), Crop Protection Cluster (CPC) at the University of the

Philippines Los Banos (UPLB).

A colorimetric Rapid Test Kit (RTK) was developed at the Pesticide Toxicology and

Chemistry Laboratory, NCPC, CPC, UPLB for organophosphate and carbamate residues (Fig 1).

The RTK was initially used as a teaching tool for conventional farmers to understand the concept

of residues and pre-harvest interval (PHI) and for them to be able to decide on the right time to

harvest.

Fig.1. Rapid test kit (RTK) for the detection of organophosphate and carbamate residues on selected vegetables developed at the National Crop Protection Center, University of the Philippines Los Banos and farmer trying to use the RTK.

74

The RTK was developed for easy and rapid detection of pesticide residues on vegetables with

tests durations of about 8-10 minutes. The RTK was tested on FPA registered organophosphate

(OP) and carbamate (CBM) pesticides on vegetable crops which are intensively applied with

pesticides. The RTK was tested on registered OP and CBM pesticides on eggplant (Solanum

melongena), yardlong beans (Vigna unuiculata), green beans (Phaseolus vulgaris), bittergourd

(Momordica charantia), pechay (Brassica chinensis), tomato (Solanum lycopersicum) and okra

(Abelmoschus esculentus).

The OP and CBM group are the commonly used/cheapest commercially available pesticides

with moderate to high toxicity. Pesticide residues are extracted using acetone concentrated and

spotted into a filter paper. After the test, the intensity of the color reaction will indicate the

concentration of the residues on the vegetable tested. The RTK is a semi quantitative

colorimetric test and completed test in a few minutes for immediate action/decision. It is cheap

(USD 0.5/test) and easy to handle. The RTK is user friendly and the procedure has been tested

and easily followed by ordinary farmers and agricultural technicians (Fig 1).

As part of expanding the applications of the RTK, it was tested on 53 MRLs on seven

commonly consumed vegetables where ten pesticides are specifically registered (Table 1).

Percentage of detection was from 19-100%. Test was sensitive to MRLs of fenthion and carbaryl

and limited sensitivity to triazophos and malathion. For the CBM test, the color of the tomato

extract interfered with the analysis of carbofuran and BPMC. However, a positive result for RTK

is still a good indication of a high concentration of pesticide residues. With the current focus of

the DA to promote organic agriculture (OA), there is a need for a rapid tool to be able to monitor

the compliance of farmer members of organic producers to ensure that no pesticide residues are

present in organic produce. For OA farmers, farmers in transition to OA, and new OA farmers, it

is necessary to monitor compliance to OA principles. The RTK may be used as a quick test of

farmer leaders and organic certifying bodies in monitoring OA farmers and help in their

regulatory work. Implementation of OA needs a technology to monitor compliance on the

avoidance of use of pesticides.

75

Table 1. RTK tested on MRLs of selected vegetables on ten FPA registered pesticides.

Pesticide No

MRLs

tested*

Range of MRLs

(mg/kg)

%

detection**

Remarks

malathion 6 0.2-8 31 Not sensitive to yardlong

beans

fenthion 2 0.1-0.5 100 Pechay and tomato

chlorpyrifos 6 0.2-1 63 Good for eggplant and

tomato

traizophos 3 0.1-0.2 19 Not sensitive to yardlong

beans and green beans

profenofos 6 0.05-10 54 Not sensitive to yardlong

beans and pechay

diazinon 6 0.05-0.5 44 Not sensitive to yardlong

beans and green beans

carbaryl 7 0.5-10 100 For all vegetables tested

carbofuran 7 0.1-0.5 87 Tomato color interferes with

detection

BPMC 4 0.3-1.5 67 Tomato color interferes with

detection

methomyl 6 0.2-5 67 Variable, good for eggplant

and green beans

*vegetables tested: eggplant, yardlong beans, green beans, bittergourd, pechay, tomato and okra

**Based on 3 persons with 3 replicates/person

MDL: 0.1 mg/kg for OPs, carbofuran,carbaryl and 1 mg/kg for BPMC and methomyl

Source: Sarmiento and Bajet (2015)

76

Table 2. Comparison between Rapid Test Kit (RTK) and Rapid Bioassay for Pesticide Residues

(RBPR).

*active ingredient registered but not on tested crops

**active ingredient not registered in the Philippines

*** tested on eggplant, yardlong beans, common beans, bittergourd, pechay, tomato and okra

The RBPR and/or RTK can be used by farmers who practice conventional, GAPs or IPM

farming to decide when to harvest, by organic and “in-transition organic” farmers to monitor

non-compliance of members and by DA technicians, traders and consumers for quick check on

the residues of vegetables. With increasing local awareness of pesticide residues and the

promotion and mushrooming of “organic farming” without any test except trust and farm visit,

the RBPR and/or RTK is expected to partly solve this need. This is cost effective compared to

monitoring using classical residue analysis. These rapid detection kits will enable the monitoring

of produce prior to and after marketing. Consumers may also opt to use RTK and/or RBPR for

personal monitoring of pesticide residues in vegetables bought from markets. Likewise, these

rapid detection kits may also be useful in regular residue analytical laboratories to minimize the

number of samples to be analyzed resulting to higher sample output and low usage of organic

solvents. In this case, only samples with known background and those which come out positive

will be analyzed.

Lastly, it can be a complimentary method for use in pesticide regulation programs of FPA,

Bureau of Agriculture and Fisheries Standards (BAFS) and Food Development Center (FDC).

This is in line with the Government’s focus on food safety.

Criteria RTK*** RBPR

Principle of test Colorimetric Enzymatic inhibition

Detection Visual,

semi quantitative

Spectrophotometer

Pesticides tested malathion, fenthion,

chlorpyrifos, triazophos,

profenofos, diazinon,

carbaryl, carbofuran,

methomyl

carbofuran, profenofos, chlorpyrifos,

methomyl, phenthoate* pirimiphos methyl*

acephate*, fenitrothion* dimethoate*,

mevinphos**, methidathion**,

metamidophos**, dichlorvos**

Time for testing

per sample

8-10 minutes 8-12 minutes

77

Monitoring of residues using classical analysis

The basis for all regulatory control is classical analysis of pesticide residues. Regulation

is based on results of locally generated residue data as part of research or regular monitoring

activities. Nationwide market basket sampling is part of monitoring residues done by BPI-NPAL

and data generated is a basis for regulatory control. If a specific pesticide residue is detected on a

vegetable crop where it is not registered, FPA will call the attention of the pesticide supplier as

part of the Pesticide Industry’s commitment on Stewardship. In cases when the residues detected

are consistently beyond MRL on specific crops, then the pesticide management practices or the

usage of the farmers are investigated. Risk management efforts are based on monitoring results

and this include dietary risk assessment using maximum residues detected or set MRLs and using

Filipino diet as basis. Dietary risk assessment is required for pesticide registration and normally

no label expansion is approved if there is > 100% of the acceptable daily intake of the pesticide

from all possible registered uses. Monitoring and risk management includes generation of local

pesticide residue data on specific crop-pesticide uses through supervised residue trials. Rice is

the crop more widely cultivated and consumes the highest volume of pesticide products but most

of the residue trials show that residues are in the husk compared to the grain. Residues are non

detectable at harvest because the last pesticide application is at milk dough stage of the rice.

Most residues on rice are due to post harvest application.

Strategies also include studies on enzymatic bioremediation of surface residues of

organophosphate insecticides (Scott et al., 2011). In the Philippines, the usefulness of the

enzyme for bioremediation was tested on mango, tomato and eggplant. Several researches on

washing with or without the aid of soap, cooking practices like broiling, boiling, frying was

found to reduce pesticide residues depending on crop surface, chemistry of pesticide, type of

formulation, and mode of action are among the researches completed to reduce dietary risks

(Bajet, 2015). The initiative of the Government to go organic farming is also one of the strategies

towards food safety, including the passing of the food safety law.

Export products like banana, pineapple, mango, okra, among others are also analyzed by

classical residue analysis for compliance to requirements related to MRLs. The results of the

analysis will be the basis for the decision to export or not and to change export destinations.

Pesticide residues are problematic especially for crops which are normally consolidated from

several farmers of different pesticide management practices like mango and okra. For mango,

Japan is the target export destination in terms of value but Hong Kong is the largest importer of

mango from the Philippines. Institutional crops like banana, pineapple and asparagus are closely

monitored in terms of pesticide and pesticide residue management by the Companies producing

these crops.

78

Environmental risk assessment and management

First line of environmental risk assessment is again through regulatory control. All pesticide

products for registration should submit data on environmental effects and environmental fate and

transport, as part of the requirements of FPA. Some pesticides result to more environmental risks

when applied to specific ecosystems, specific times of application or proximity to waterways and

other non-target organisms. Risk management involves knowledge of these situations and to

relate to toxicity like lethal dose resulting to 50% mortality (LD50) values of non-target

organisms and relate to the measured or expected environmental concentrations.

Researches related to environmental risk monitoring and assessment is based on classical

analysis of pesticide residues on targeted ecosystems and organisms. Several studies were done

at NCPC, UPLB and other institutions in the country. Environmental risk is greater for rice

cultivation since the probability of off site migration is higher due to the irrigation canals which

eventually find its way to bodies of water. Rice is also the largest consumer of plant protection

products due to the wide area of cultivation although the intensity of usage is lower than high

value crops. The pesticide application and resulting residues were documented by Bhuiyan and

Castaneda (1995) and Tejada et al., (1995) in the early `90s. Recently, Elfman et al. (2011)

reported that for 34 samples analyzed, 16 samples collected from rice ecosystem exceeded the

Swedish guidelines for lambda cyhalothrin, cypermethrin and deltamethrin in San Francisco,

Leyte Philippines (Table 3). Mollusicides, also used in rice farming for the control of the Golden

Apple Snail, could also be a source of contaminants in shellfish in the coastal regions. Tanabe et

al., 2000 measured total butyl tins (BTs) in green mussel with concentrations ranging from non

detectable to 787 ng/g and an average of 133 ng/g of the 11/13 samples found to contain BTs.

In Benguet Province, the vegetable bowl of the Philippines, Del Prado-Lu (2010) detected

chlorpyrifos in 2% of the water sampled and 43.5% of the soil samples analyzed contain

endosulfan sulfate, chlorpyrifos, profenofos, cyhalothrin, cypermethrin, endosulfan and

chlorothalonil residues. This is indicative that the soil is really the ultimate sink of pesticide

residues. Endosulfan is also already banned for use in the Philippines, which explains the

presence of the metabolite endosulfan sulfate.

Varca (2012) monitored the off site migration of pesticides in the Pagsanjan-Lumban

watershed which is the fresh water source of Laguna Bay, the second largest freshwater Lake in

Southeast Asia. This watershed runs through vegetable and rice production areas and an average

of 0.85 ug/L malathion was detected in 2008 and 0.113 ug/L in 2009. A concentration of <1.0-

15.4 ug/L prefenofos was also detected in 2008 (Table 3). The maximum measured

concentration of malathion and profenofos in water was related by Bajet et al. (2012) to toxicity

to aquatic non target organisms.

79

Table 3. Selected measurements of pesticide residues in Philippine environment.

Ecosystem (Place) Environmental

substrates

analyzed

Results Reference

Rice (San

Francisco, Leyte)

water lambda cyhalothrin (0.006

ug/L); cypermethrin and

deltamethrin (0.0002 ug/L)

Elfman et al.,2011.

Agricultural Water

Management 101:

81-87

Vegetable (La

Trinidad, Bugias

and Atok, Benguet

Province)

water

soil

1/49 (0.07 mg/L

chlorpyrifos)

34/78 samples with residues

(endosulfan sulfate,

chlorpyrifos, profenofos,

cyhalothrin, cypermethrin,

endosulfan, chlorothalonil))

Del Prado-Lu. 2010.

Archives on

Environmental

Contamination

Toxicology 59:175–

181

Vegetable and rice

(Pagsanjan-

Lumban

Watershed, Laguna

de Bay)

water 0.85 ug/L average malathion

2008 and 0.113 ug/L in 2009

<1.0-15.4 ug/L prefenofos in

2008

Varca, 2012.

Agricultural Water

Management 106:

35-41

Environmental

Monitoring (Metro

Manila, Cavite,

Leyte, Capiz,

Bataan)

Green mussel Total butyl tins (range: non

detectable to 787 ng/g with

average of 133 ng/g)

Tanabe et. al., 2000.

Ocean and Coastal

Management 43:819-

839.

Ecotoxicologcal

impact (Pagsanjan-

Lumban

Watershed, Laguna

de Bay)

Tilapia

fingerlings

Shrimps

Duckweed

Environmentatl

concentration of profenofos

and malathion with hazard/

impact on local shrimps

species of Laguna de Bay

Lesser risk of herbicides to

duckweed

Bajet et al., 2012.

Agricultural Water

Management 106:

42-49.

Environmental impact management includes measuring the toxicity of pesticides on local

species like fish, shrimps, and duckweed, among others. Toxicity and toxicity index were

measured by Tejada et al. (1993) on Tilapia fingerlings. Toxicity of non-target aquatic organisms

80

were related to the measured environmental concentration in the Pagsanjan-Lumban watershed

which is the main tributary to Laguna Lake and passes through agricultural areas mainly rice,

vegetables, plantation crops and ornamentals (Bajet et al. 2012). The maximum measured

environmental concentration of profenofos and pyrethroids were greater than the 48h LC50 of

local shrimp Macrobrachium sp. indicating that this species is sensitive to the use of these

pesticides in the watershed (Table 3). The local duckweed Lemna sp, used as a fish food for

Tilapia sp. is also used to measure of the impact of herbicides on the aquatic environment (Bajet

et al., 2010)

CONCLUSION

The Philippine Government’s effort towards food and environmental safety in terms of pesticide

residues is focused on regulatory control of a number of Government agencies and passing of

laws related to food safety and organic agriculture in partnership with stakeholders. In support,

monitoring activities through research as well as development of rapid test kit and pilot testing of

RBPR are efforts to monitor residues of local markets and farms. Environmental monitoring is

also part of researches of Government agencies and Institutions to be able to gain knowledge in

management of these risks. Lastly, with limited funds for food monitoring, the effective way to

increase food safety is to promote organic agriculture, strengthen programs that can guide

farmers on good farming practices, alternative methods for pest control like IPM, GAP, use of

plants with pest repellent properties, recognizing beneficial insects, know about pesticide

residues and judicious/proper use of pesticides, all of which can reduce violative residues on

food resulting to minimal dietary and environmental risk.

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Bioremediation of Pesticides. A Case Study for the Enzymatic Remediation of

Organophosphorous Insecticide Residues. In Pesticide Mitigation Strategies for Surface

Water Quality. ACS Symposium Series, 1075: 155-174.

Tanabe, S., M.S. Prudente, S. Kam-atireklap, A. Subramanian. 2000. Mussel watch: Marine

pollution monitoring of butyltins and organochlorines in coastal waters of Thailand,

Philippines and India. Ocean and Coastal Management, 43:819-839.

Tejada, A.W., L.M. Varca, S.M. F. Calumnpang and C.M. Bajet. 1998. Enzyme inhibition and

other rapid techniques for pesticide residue detection. In seeking agricultural produce free of

pesticide residues. ACIAR Proceedings No. 85: 223-228.

Tejada, F.R. 1995. Development of a Rapid method for the detection of carbamate and

organophosphate insecticides using enzyme inhibition technique. University of the

Philippines Undergraduate thesis, College, Laguna Philippines.

http://pubs.acs.org/isbn/9780841226562

http://pubs.acs.org/isbn/9780841226562

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Tejada, A.W., C.M. Bajet, E.D. Magallona, M.G. Magbanua, N.B. Gambalan and L.C. Araez.

Toxicity and Toxicity Indices of pesticides to some fauna of the lowland rice fish ecosystem.

Philippine Journal Agriculture 76:373-382.

Tejada, A.W., L. M. Varca, S.M.F Calumpang, P.P. Ocampo, C. M. Bajet, E.B. Paningbatan,

J.R.Medina, V. P.

Justo, C.D.L. Habito, M. R. Martinez and E.D. Magallona. 1995. In Pingali PL & PA Roger (eds)

Impact of Pesticides on Farmer Health and the Rice Environment. Kluwer Acad. Pub.,

Massachusettes, USA and Dordrecht, Netherlands. 149-180.

Varca, L.M. 2012. Pesticide residues in surface waters of Pagsanjan-Lumban catchment of

Laguna de Bay,

Philippines. Agric. Water Management 106: 35-41.

SUMMARY

The Philippines in its effort to promote food safety has mainly focused on Government

regulatory control through pesticide registration, monitoring, training and partnership with

stakeholders. There is limited monitoring of local markets but data generated is used for

regulatory control. Efforts to augment pesticide residue monitoring activities are a consequence

of research activities, stakeholders and Industry efforts using classical pesticide residue analysis

and rapid detection tools. Environmental safety is also a concern with researches generated

through measuring the impact of agricultural activities on water, soil, and other environmental

compartments. Ecotoxicological studies on the impact on non target organisms is also

determined by measuring toxicity and relate data to expected and/or measured environmental

concentrations of pesticide residues.

83

Session II : Novel technologies in practical: Agrochemicals residues

reduction and food safety improvement

84

APPLICATION OF ULTRASOUND TECHNOLOGY FOR

AGRICULTURAL PRODUCT IMPROVEMENT AND ENVIRONMENTAL

RENOVATION

Nakao NOMURA1, Takumi Sasaki

2,

Kanda Whangchai3, Sarunya Pengphol

4 and Niwooti Whangchai

5

1 Global Commons Organization (Faculty of Life and Environmental Sciences), University of

Tsukuba, Japan 2 Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan

3 Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 4 Department of Agriculture, Faculty of Agricultural and Industrial Technology, Nakhon

Sawan Rajabhat University, Nakhon Sawan, Thailand 5 Faculty of Fishery Technology and Aquatic Resources, Maejo University, Chiang Mai,


ABSTRACT

Ultrasound radiation has been applied for analytical observation of lake sediment as well as

improvement of aquaculture and agricultural products. Optimum conditions for each application

was determined by using small ultrasound device where different frequency, input power can be

adjusted. Under optimum condition, lake sediment can be observed with information of textures

of bottom soil, which could indicate contents of organic matters and degree of pollution. For the

improvement of sea water aquaculture products and fresh water aquaculture products, one of the

major programs in fresh water aquaculture was applying used to ultrasound technology to

minimize musty odor. As a result, growth of phytoplankton which produces musty odor

compounds was able to be controlled by destroying gas vacuole in it cells and musty odor

compounds were able to be decomposed by applying ultrasound radiation coupled with

photocatalytic material. For decomposition of residual agri-chemicals in agricultural products,

ultrasound radiation coupled with ozonation was applied and results showed that ultrasound

radiation coupling with ozonation was able to achieve removing more than 75% residual agri-

chemicals.

Keywords: Ultrasound radiation, Phytoplankton, Musty odor compounds, Agri-chemicals,

Decomposition


85

INTRODUCTION

Ultrasound technology has been widely utilized for 2 purposes. First purpose is to obtained

ultrasound signals from target substances through reflection echoes: a) Imaging the target

substances with minimum physical damage or stress on the substances (Nakao et al., 2003); b)

Medical application and clinical uses: devices to observe some parts of animal bodies such as

internal organs, tumor masses or fetus have been developed. C) Fish finder: fish finder has been

used to detect school of fish for sea-catch fishery activities. Second purpose is by utilizing

kinetic power of ultrasound for physico-chemical reaction to improve quality of target

subsntances (Takumi et al., 2015, Sarunya et al., 2011, Kanda et al., 2013). For this purpose,

ultrasound technology was applied to a) Observe lake sediment, and to b) Removal or

decomposition of chemical substances causing quality issues in aquaculture and agricultural

crops.

ANALYTICAL OBSERVATION OF LAKE SEDIMENT

Many of enclosed water bodies in Japan are suffering from water quality deterioration due to

eutrophication. Although local and central government has been improving water quality in

rivers or creeks flowing into these water bodies, water quality in the water bodies is still in same

situation in most of the cases, meaning that internal nutrition load is a main cause of water

quality deterioration. Through several analysis and survey to clarify source of internal load such

as nutritional input by rainfall, birds, fishery industries, it was found that main part of internal

nutrition load in enclosed water bodies are from nutrient release from sediment in many cases.

However, monitoring and analysis of sediment in large scale takes lots of time and cost. This is

because current sediment analysis depends on laboratory analysis of the sediment sample taken

from the water bodies by using core-sampler or Ekman bottom grab sampler. Therefore, the

ultrasound technology was applied to detect the bottom of the water body and analyze the

textures of sediment, particularly softness and particle sizes of the bottom soil. The team aim to

determine optimized ultrasound condition for the testing site Kasumigaura Lake. When

observing bottom soil of Lake Kasumigaura, it was found that the optimum frequency is 20- 40

kHz. The same methodologies also applied in different locations in Japan (Mikawa Bay in Aichi

prefecture, Lake Senba in Ibaraki prefecture) and were both found effective.

86

IMPROVEMENT OF FRESH WATER FISHERY PRODUCTS BY

MINIMIZING MUSTY ODOR COMPOUNDS

Nile Tilapia is one of the successful and profitable aquaculture products in Thailand. Since

Tilapia can be cultured both in sea and fresh water, it has been widely cultured in many regions

of Thailand. Nowadays, inland farming of Tilapia is increasing due to coastal regions farmers

tend to shift from Tilapia to prawn aquaculture, which bring higher profit. Tilapia cultured in sea

water often encounter problems of pathogenic halophilic bacteria outbreak. And inland Tilapia

culture has serious musty odors problem that derived from some metabolic products of

phytoplankton, those are abundant in fresh water Tilapia farms. In this study, ultrasound

technology applied to solve musty odor problem in 2 ways. First was to minimize growth of

phytoplankton by destroying gas vacuole in the cell of phytoplankton. The optimum condition of

ultrasound to treat phytoplankton was determined (200kHz) and the treatment was applied. Fig. 1

illustrate photographs of phytoplankton before (0 min) and after (20, 60 min) ultrasound

treatment, showing that majority of the phytoplankton sank down to the bottom of volumetric

cylinder due to disruption of gas vacuole. Second was to decompose musty odor substance such

as geosmin and MIB (2-methyl sobornol). As shown in Fig. 2, results indicated that with

ultrasound radiation only (sonolytic treatment) is not enough for full-decomposition of geosmin

(Result with MIB was in almost same trend). This was confirmed by determining concentration

of residual geosmin or MIB by GC-MS. However, when applying TiO2, a photo-catalytic

material, together with ultrasound radiation (sono-photocatalytic treatments) effectively

decomposed musty odor substances. This indicated ultrasound radiation coupling with other

oxidation materials could assist the decomposition process.

Fig. 1. Photographs of phytoplankton in a fresh water pond after treatment of ultrasound

radiation

Treatment

Time

(minutes)

0

bottom Surface (1) Surface (2)

20

60

87

Fig. 2. Geosmin removal rate after photocatalytic, sonolytic and sono-photocatalytic treatments.

IMPROVEMENT OF AGRICULTURAL PRODUCTS BY

REMOVING RESIDUAL AGRI-CHEMICALS

Ultrasound radiation was applied to remove agri-chemicals in agricultural crops. Similar result as

above mentioned “musty odor substance elimination” experiment shown that single ultrasound

radiation treatment also not enough to remove target chemicals in agricultural crops.

Ultrasound has effect to wash and polish up solid surface of several substances; it was expected

to enhance detachment of the agri-chemicals from crop surfaces into liquid phase by applying a

chamber equipped with ultrasound transducers (Figure 3) filled with water and agricultural crops

contaminated with target agrichemicals. Ethion is widely applied to fruit tangerine and

Chlorpyrifos widely applied to bird chili. The objective of this experiment is to remove agri-

chemicals Ethion and Chlorpyrifos. Results showed that ultrasound radiation coupling with

ozonation was able to remove more than 75% of Ethion and Chlorpyrifos.

0

10

20

30

40

50

60

70

80

90

100

Photocatalysis Sonolysis Sono-photocatalysis

Ge

os

min

re

mo

val ra

te (

%)

Sample 1 Sample 2 Sample 3 Sample 4

Treatment

88

Fig. 3. Photographs of ultrasound chamber for treatment of agricultural crops

CONCLUSION

In this study, ultrasound radiation was applied for environmental renovation and improvement of

aquaculture and agricultural products. The optimum condition for each ultrasound radiation

application was determined. It was found that ultrasound radiation coupled with photocatalytic

materials or ozonation was effective for decomposition of musty odor compounds and residual

chemicals in crops. The information obtained in this study is expected to contribute for further

development of commercial devices towards environmental renovation or products improvement

in industrial scale.

ACKNOWLEDGEMENTS

Authors would like to offer special thanks to Honda Electronics Co. Ltd. To provide

experimental devices and support most of research studies. Authors also would like to offer

special thanks to Sydney Water to provide experimental site and technical support for the

experiments in Sydney Olympic Park.

89

REFERENCES

Nakao N. et. al., 2003, A novel device to monitor bottom soil of water basin using

ultrasound technology. Japanese Journal of Water Treatment Biology 39(3) 139

Takumi S. and Nakao N. et. al., 2015, Removal of Off-flavor compounds and

Cyanobacteria in Fish Ponds Using Sonophotocatalysis, The

9th Fisheries Academic Conference, MaejoUniversity, Chiang Mai

Sarunya P. and Nakao N. et al. 2011, Effect of Frequency and Sonication Time of

Ultrasonic Irradiation in Combination with Ozone on Postharvest Chlorpyrifos

Residue Reduction in Fresh Bird Chilli (Capsicum frutescens Linn.) Fruits.

Agricultural Science Journal 42(1) 236, 2011

Kanda W. and Nakao N. et. al. 2013, The effects of ultrasonic irradiation in combination

with ozone on the reduction of residual ethion of tangerine (Citrus reticulata Blanco

cv. Sai Nam Pung) fruit after harvest Agricultura

90

USING SANITIZER AND FINE BUBBLE TECHNOLOGIES

TO ENHANCE FOOD SAFETY

Warapa Mahakarnchanakul1,

Pitirat Klintham1, Sasitorn Tongchitpakdee

1, Wannee Chinsirikul

2

1

Department of Food Science and Technology, Faculty of Agro-Industry, Kasetsart University,

Bangkok, THAILAND 2

National Metal and Materials Technology Center, National Science and Technology

Development Agency, Ministry of Science and Technology, Pathumthani, THAILAND

email: [email protected]

ABSTRACT

Fine bubble is defined as the small bubbles with a diameter ranging in micro and nanoscale.

The sizes of micro bubble are smaller than millimeter and have distinctive properties. The

applications of micro bubbles technology has been successfully proved in waste water treatment.

More application was investigated in washing process to reduce microorganisms and pesticides.

The effectiveness of micro bubble water to reduce E. coli and Salmonella contamination on six

Thai fresh vegetables during washing step was conducted. A preliminary study was performed

on artificially contaminated coriander, Marsh mint, asparagus, okra, lemongrass and ginger.

Washing samples for 15 minutes by micro bubble water at flow rate of 4.5 L/ min successfully

reduced both pathogens in these tested vegetables. Second experiment was done to investigate

the combination of micro bubbles with sanitizers, sodium hypochlorite, acetic acid and citric

acid in washing Chinese kale. The results showed the promising methods to reduce microbial

load compared to normal washing, although no significant among the different at concentration

levels. These techniques have potential to apply in washing steps to enhance the safety of food,

particular in fresh produce process.

Keywords: Micro bubbles, washing produce, foodborne pathogen, sanitizers, produce safety

91

INTRODUCTION

Microbial contamination in fresh produce

Recently, there has been an increasing consumption of fresh produce worldwide due to the

concerning of consumers on their health benefits. Meanwhile food safety problems particularly

the pathogenic microorganisms linked to contaminated fresh produce have been repeatedly

reported (Kirezieva, et al., 2015). Foodborne pathogens could contaminate on fresh produce at

many steps along the production from farm to table at point of consumption. The contamination

of foodborne pathogen can arise from environmental, animal or human sources (Gil, et al.,

2009), thus, the prevention and control to the contamination on fresh produce has been focused

(Fan and Sokorai, 2015).

In terms of food safety not only the significant microbiological contamination issues

concerning as threats to health, but also the issues associated with the market of fresh produces,

and in particularly international trade. In 2005, 32 out of 244 herbs imported from non-EU

countries, sold in London, UK, were contaminated with Salmonella, those including four

varieties of basil grown in Thailand. Again in 2006, 5 out of 298 fresh herbs in the UK including

coriander, curry leaves, and holy basil that imported from India and Thailand were contaminated

with Salmonella (Elviss et al., 2009).

The majority of pathogens that implicated in fresh produce outbreaks are enteric micro-

organisms that originated from the gut of warm-blooded animals (Xuetong et al, 2009). For

example the enteric pathogens such as Salmonella spp. Among these enteric bacteria, both genus

Salmonella and Escherichia coli O157:H7 are the major pathogens contributing to outbreaks of

foodborne illness associated with fresh produce (Delbeke et al., 2015; Olaimat and Holley, 2012)

Factors contributing to the contamination in fresh produce

There are numerous factors leading to microbial contamination, human pathogens can be

introduced to fresh produce production systems through animal feces, from wildlife or livestock

or by the application of manures as fertilizers (Beuchat, 2002; Xuetong et al, 2009). Number of

the presence microorganisms differed depending on the type of produce, agronomic practices,

geographical area of production, and weather conditions. Other than the hygienic conditions,

pre-harvesting, post-harvesting, transportation, further processing and handling of fresh produce

can also significantly influence on the microbiota pattern (Ramos et al., 2013).

Leaf topography is an importance factor for microbial adhesion. The surface roughness of

the leaves, crack in the cuticle and other damages are often sites at which bacteria colonize. Leaf

stomata provide protective niches for the bacteria. E. coli O157:H7 cells have been found that

cells could adhere better to cut lettuce leaf surfaces than intact or normal lettuce leaf surfaces

(Seo and Frank, 1999). Many reports showed human pathogen can enter stomata and cut edges

of fresh produce (Berger et al., 2010; Golberg et al., 2011) as well as the ability of E. coli

O157:H7 and Salmonella to internalize or infect the vascular system of growing plant (Olaimat

and Holley, 2012).

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Washing fresh produce

Fresh produces washing is a significant step for removing soil, debris, and improving the

appearance of the commodity. Washing also to lowering the produces’ temperature to limit the

development of physiological changes. Moreover, washing helps to reduce the microbial load on

the surface which has impact to quality, shelf-life, and safety of the produces (Herdt and Feng,

2009). Washing produces with water or sanitizing agents could help to remove pesticide

residues (Sapers, 2014).

In contrast, washing can transfer microbial contaminants to wash water and also to other

uncontaminated raw materials, as well as to the processing equipment (Gil et al., 2009).. Adding

sanitizing agents to wash water significantly reduce the population of bacterial cells, and thus the

risk of cross-contamination (Sapers, 2014). Washing with sanitizers assist to avoid cross-

contamination between clean and contaminated product and significant enhance produce hygiene

(Gil et al., 2009). Typically a microbial load of fresh produce can be reduced 1 to 2 logs by

washing and disinfection. However, washing is restricted to treat the microorganisms on the

surface and internalized microorganisms of fresh produces (Lee et al, 2004).

Not all washing methods and washing solutions are effective (Olaimat and Holley, 2012), the

success of washing depends on many factors: type of washing, exposure time, the concentration

of sanitizing agent, pH, temperature as well the target microorganisms, the characteristics of

produce surface, the attachment of cell to produce surface, the formation of resistant biofilms and

the internalization of microorganisms (Allende et al., 2008). The challenges of choosing

sanitizing treatment are how to reach pathogens on the surface and in subsurface areas of fresh

produce in an active formand and at the same time compromise the quality of fresh produce after

treatment (Beuchat, 2004).

Sanitizing agents for fresh produces sanitation

Three factors could influence the efficacy of disinfection need to be noted. Firstly, sanitizing

agents including pH, temperature, and water quality; secondly the availability of disinfectant in

water which are bound to the inorganic and organic compounds; and lastly the accessibility of

sanitizing agents to the target microorganisms (Barbeau et al., 2005).

The chosen of chemical sanitizing agent during the washing disinfection step is a critical step

for keeping quality and safety of fresh produces. Ideal sanitizing agent should show sufficient

level of antimicrobial activity, and be a negligible effect on the sensory quality of the product

(Allende et al., 2008). Most of the common type of sanitizers are oxidizers, working by

increasing the oxidation potential in the water system, and are considered to be the Generally

Recognized As Safe (GRAS).

Chlorine (Cl) is the most widely sanitizing agent that used to sanitize fresh produce (Sapers,

G. M., 2014). It is a very potent disinfectant with powerful oxidizing properties. However an

antimicrobial activity of chlorine depends on the pH of wash solution, for the best compromise

of activity and stability of hypochlorous acid (HOCl) should be potentially control and maintain

at the pH of wash water between 6.5 and 7.5. Currently in the fresh produce industry used

Sodium hypochlorite (NaOCl) to disinfectant produce at a concentration of 50-200 mgL-1

and a

contact time of 1-2 min (Beuchat, 1998;Fan and Sokorai, 2015;Goodburn and Wallace, 2013).

Another was reported at 50-150 mg/L (Martínez-Hernández et al., 2015). However the

chlorination water containing free chlorine between 20 to 200 mg/L in sanitizing solution cannot

93

eliminate pathogens completely, the reduction of 1 to 3 log CFU/g are common (Aruscavage et

al., 2006).

Alternative sanitizing agents used for fresh produce such as electrolyzed oxidizing water,

chlorine dioxide, ozone, peroxyacetic acid, acidified sodium chlorite or peracetic acid have been

studied (Alvaro et al., 2009;Alwi and Ali, 2014;Artés et al., 2009;Gil et al., 2009;Gómez-lópez,

2012;Goodburn and Wallace, 2013;Graça et al., 2011;Hao et al., 2015; Herdt and Feng,

2009;Olaimat and Holley, 2012;Ramos et al., 2013)

Electrolyzed oxidizing water (EO water or EOW) is an emerging decontamination

technology. The EOW system was developed in Japan in 1992 and EOW has been attracted

much attention in recent years. It is not only effective solution for inactivation of

microorganisms but also is considered as environmentally friendly chemical (Cheng et al., 2012).

The role in disinfection mechanism of EOW consists of high oxidation-reduction potential

(ORP), pH and available chlorine content (ACC) ae well as the OH radical in EOW (Hao et al.,

2015;Hao et al., 2012;Stan and Daeschel, 2005;Stan et al., 2005). The acidic EOW which had a

low pH (2.5 to 3.5), high ORP (1000 to 1200 mV), high dissolved oxygen and free chlorine (30

to 90 mg/L) has been regarded as an effectively antimicrobial activity (Ding et al ., 2015; Huang

et al., 2008).

Chlorine dioxide (ClO2) is a biocide with 2.5 times the oxidation capacity of chlorine

(Rodgers et al., 2004). Chlorine dioxide is highly stable and less corrosive than ozone and

chlorine (Joshi et al. , 2013). It can apply as an aqueous solution, or gas phase (Sapers, 2014).

Moreover, ClO2 is not active to organic compounds (Han et al., 2004; Lee et al. , 2004). FDA

approved the formulation of ClO2 as an antimicrobial agent in water (aqueous ClO2)

(21CFR173.300) and can use ClO2 for wash fresh fruit and vegetables. However the residual of

ClO2 in final produce should not over than 3 mg/L of ClO2 and suggestion to rinse with potable

water after treatment with ClO2 (López-Velasco et al., 2012; Sapers, 2014).

Emerging technology: Fine Bubble Technology

Fine bubble is defined as the small bubbles with a diameter ranging in micro and nanoscale.

Microbubbles are the small bubbles with diameter between 10 to 50 µm and decreasing in size

and lastly disappear under water (Hideki, 2014; Parmar and Majumder, 2013; Takahashi, 2005;

Takahashi et al., 2007), while the nanobubbles is evensmaller bubbles with diameter less than

200 nm (Agarwal et al., 2011). Stabilized nanobubbles are created when microbubbles are

collapse in aqueous electrolyte solution (Masayoshi, 2014).

Microbubble composts of three main components, gas phase, shell material, and aqueous or

liquid phase. The gas phase is referred to the gas inside the bubbles which may be single gas or

the combination of gases. The second is the shell material, an aqueous phase surrounding the gas

phase. The formation of the bubbles and the mechanical properties of microbubbles depend on

the property of shell material. The last important component is aqueous phases which are the

liquid or combined solution surrounding the shell material (Parmar and Majumder, 2013). These

three components mainly contribute to the properties of microbubbles.

Several interesting characteristics of microbubbles comparing with a millimeter or

centimeter-sized bubbles are described. Although the microbubbles are defined as the bubbles

having diameter in order of micrometer (µm) (Hideki, 2014; Parmar and Majumder, 2013),

however the range of the bubbles diameter can be varies. Usually the microbubbles have a slow

94

rising speed. Besides, the Reynolds number of microbubbles is nearly 1 and its shape is

spherical (Hideki, 2014).

Microbubbles could reduce frictional resistance, therfore the coefficient of friction decreased

with increased in volume fraction of microbubbles (Hideki, 2014; Parmar and Majumder, 2013).

The internal pressure of bubbles depend on the bubbles diameter and surface tension of bubbles.

Decreasing of bubbles diameter caused the increasing of internal pressure inside of the bubbles,

that may describe by the Young-Lapace equation. The occurrence of this phenomenon can be

explained as increasing the gas dissolution will increase the mass transfer rate due to driving

force of gas dissolution in the internal pressure (Parmar and Majumder, 2013). Lastly,

decreasing microbubbles in water affect an increasing of the large gas-liquid interfacial area and

the changes of the physical properties of water (Hideki, 2014).

Microbubbles have been suggested that they possses a negative charged on their surface

(Hideki, 2014; Takahashi, 2005). The studied of the zeta potential of microbubbles in aqueous

showed the zeta potential of microbubbles in distilled water was -35 mV (Takahashi, 2005). The

zeta potential of the particle is a physical property that is possessed by any particle in suspension

and emulsion. Usually the zeta potential of emissions used to predict the stability of suspension

and emulsion stability. Likewise, the fine bubbles in the solution similar characteristic like the

particle in suspension and having a significant negative potential tend to repel the other particle

(Parmar and Majumder, 2013).

Similarly to the nanobubbles, they are tiny bubbles with the diameter in nanoscale and

smaller size than microbubbles. It is invisible in solution while microbubbles are visible. Before

the generation of microbubbles water is clear and its change from the clear solution to cloudy

and milky after the generating the microbubbles (Masayoshi, 2014 ; Li, 2007). Then the smaller

bubbles are shrunk faster and disappear almost instantaneously, and the nanobubbles are created

after the microbubbles collapse. The half-life of nanobubbles depends on the water condition,

many researchers are trying to clarify the mechanism of nanobubbles stability but the difficulty is

the measurement the properties of these tiny particles (Tsuge, 2014)

There are many fields application research of microbubbles and nanobubbles such as

environmental field, industrial field, agricultural field, medical and food industry due to the

different function properties. Recently bubble technology is applied in waste water treatment

particularly in food industries, and thus this technology influences the quality, production

efficiency or cost of food products. In food process the development of foods using microbubble

(MB) or micro-nano bubble (MNB) technology is expected to obtain a new characteristic

because the quantity and size of the food bubbles may contribute to their appearance, physical

properties and texture of the foods. The application extends to control over the growing and pre-

postharvest physiological properties of fresh produce, sterilization of food or equipment, besides

the treatment of public water and wastewater (Xu et al., 2014).

Newly developed or quality-improved food/food materials are expected to be produced by

microstructure control. Regarding to the process of cream production, Kukizaki (2009)

described that controlling the surface characteristics of Fine bubbles (Micro- and nano-Bubbles)

will contribute to improving the stability of the emulsion. Apart from that fine bubble is

expected to be used as a delivery system of functional or aroma components in foods and used as

a carrier of drug or antimicrobial agents to maintain food hygiene or safety. Micro bubbles

treatments were applied for sterilization, some biocidal gases are often incorporated, therefore

with the combined effect of ozone bubbling and ozone nanobubble treatment have been shown

significant microbial efficiency. Moreover the combination microbubbles with ozone gas

95

effectively reduced the pesticide residual on fruits and vegetables ( Ikeura et al, 2011a; 2011b

and 2013)

The application Micro/Nano bubble on reducing pesticides in fresh produce

In 2011, Ikeura and coworker compared the efficacy of two types of ozone microbubbles

generator, the decompression-type and the gas-water circulating-type, to remove the residual

pesticide fenitrothion in lettuce, cherry tomatoes and strawberries. The decompression type

produces a sufficient amount of gas that dissolved in water under a 3-4 atmospheric pressure to

cause a supersaturated condition. Under such a condition, supersaturated gas is unstable and

when escapes from the water generating a lot of air bubbles, which are microbubbles. While the

gas–water circulation type, gas is introduced into the water vortex, and the formed gas bubbles

are broken into microbubbles by breaking the vortex. They found that the concentration of

dissolved ozone was higher in the decompression type solution than in the gas–water circulation

type solution, although the concentration of dissolved ozone decreased gradually with time.

Washing with ozone microbubbles water effectively removed residual fenitrothion in all

vegetable samples (Fig. 1), and the decompression type was shown to be better. Hydroxyl

radicals that are generated by the collapse of ozone microbubbles in solutions are highly

effective at decomposing organic molecules (Ikura et al., 2011a), resulting in pesticides

degradation.

Reduction rate of fenitrothion on cherry tomatoes was slightly different from the other

vegetables because the dissolved ozone and hydroxyl radicals could hardly penetrate through the

thick pericarp of the cherry tomatoes. While strawberries fruit that have a rougher surface and

larger surface area than cherry tomatoes, fruits can contact with ozone efficiently, therefore

removing fenitrothion are easily compare to tomatoes. The difference in the pesticide-removing

effect between the decompression type and the gas–water circulation type may be caused by the

difference in the size and the number of the bubbles (Ikura et al., 2011b).

Later Ikura and co-worker (2011b) found ozone microbubbles (OMCB) produced by

continuous microbubble generator could retain the concentration of dissolved ozone at 2.0 ppm

(Fig 2.), resulting in effectively removal of fenitrothion better than the ozone millibubble

(OMLB). Continuous microbubble generator removes the pesticide better than the bath

generation or non-continuous generating. Later different type of pesticides were investigated by

Ikeura et al. (2013), fenitrothion is represented as organophosphate insecticide while benomyl is

carbamate fungicide. Since allowable pesticides residues in food have been implemented in

Japan and other countries, including persimmon wrapping leaves for special sushi in Nara, Japan.

Red and green persimmon leaves were chosen as a sample model.

The pesticide-spiked persimmon leaves were subjected to the treatments by different micro

bubble generation methods. Ozone concentrations in water varied from 0.2, 0.5, 1.0 and 2 ppm,

at immerge times 5, 10 or 15 min, were subjected to leaf samples. The residual of fenitrothion

on persimmon leaves were decreased when increased washing time from 5 to 15 min, and again

continuous generating the microbubbles was shown better effective removal of the fenitrothion

than batch generating, likewise the reduction of benomyl on these leaves. No effect of bubbling

or continuous microbubbles during treatment to the color and the pulling strength of Persimmon

leaves were found (Ikura et al., 2011b).

96

Experiment I : Removal of foodborne pathogen in Vegetables using combination of

sanitizers and microbubbles. (Mahakarnchanakul et al., 2010)

Six types of popular Thai fresh produce were selected and classified into three types;

leafy type: coriander (Coraindrum sativum) and peppermint or Marsh mint or Thai mint (Mentha

arvensis); Pod and stem type: asparagus (Asparagus officinalis) and okra (Abelmochus

esculentus); Root type: ginger (Zingiber officinale) and lemongrass (Cymbopogon citrates).

All fresh produce were stored at 5°C and placed on the laboratory bench for 30 min, then

selected similar size and shape. The 18 h suspension of Salmonella Hvittingfoss and Escherichia

coli were prepared for inoculum, the final load on each vegetable type was expected to be 103-

104 CFU/ml. No wash before inoculating. Then, vegetables were inoculated by adding culture

suspension in Polyethylene bag, manual shaking for 2 min, placing under a laminate flow cabinet

to let them dry for 30 min. Samples were analyzed for initial loads.

Micro bubble water was prepared by circulating the tap water through a generator in sterile

plastic tank (5L). MB generator was obtained from Shigen Kaihatsu Co., Ltd., Japan. Five

hundred grams of each sample was submerged in 5 L of micro bubble water (at flow rate of 4.5

L/min) by ratio 1:10 and, agitating for 15 min. Then vegetables were drained and dried under

laminate flow cabinet at 20± 3oC (65-70% relative humidity) for 30 min before microbial

analyze.

Total viable counts were determined on Plate Count Agar (PCA) while E. coli was counted

on McConkey agar, likewise S. Hvittingfoss on Xylose lysine desoxycholate (XLD). Plates were

incubated at 35 °C for 24 h before enumeration the survivors (Figure 4). All samples were

enumerated by rinse test to avoid the natural antimicrobial from smashed vegetables.

The fragile leaves of coriander and Marsh mint are easily damaged from the harsh washing

process. Washing by using MB water showed distinctively good appearance and fresh green

leaves due to dissolved oxygen during bubbles generating. The reduction of microbial

population by washing with micro bubble water was better than normal washing with tap water.

MB treatments could reduce microflora on coriander by 87.6% log reduction, while tap water

could reduce microflora load about 56% log reduction. While washing Marsh mint showed

microbial reduction by tap water and micro bubbles treatment as 95 and 80 % reduction,

respectively (Figure 3A and 3B).

Thai asparagus and okra were known as major exporting produce particularly for Japan

market. Washing with MB water and tap water could reduce natural microflora on asparagus

and okra. The reductions of total microbial on asparagus and okra using MB water were 99.6

and 98 % compared to washing with tap water, which gave reduction less by 92 and 95 %.

(Figure 3C, and 3D). The MB showed the high efficiency in reducing normal flora in asparagus

and okra by 1.7-2.4 log CFU/g.

In case of root type, lemongrass and ginger, normally the natural load was high as 6-7 log

CFU/ml due to the nature of root plants. Rough and cracked surfaces of ginger prone to be

heavy contaminated with soil and difficult to remove soil, resulting in high microbial population

and E. coli presence. Fig 3E and 3F showed the remarkably reduction of microflora on

lemongrass and ginger samples. Before washing with MB, microflora on lemongrass and ginger

were 5.8 and 6.8 log CFU/ml and after washing population was reduced to 4.3 and 5.8 log

CFU/ml (97 and 89% reduction). Washing lemongrass and ginger using tap water showed

slightly less reduction by 93 to 94%.

97

In conclusion the number of total bacterial count (microflora) deposited on surface of tested

vegetables was contaminated from 105 to 10

7 CFU/ml, microbial population was varies

depending on the different physiology of plant. Each type of vegetables has different in texture

and level of microbial contamination, besides the presence of small fur on Marsh mint leaves, or

the crevices of surfaces of ginger resulting in the possible habitat. The chance of soil contact

during harvesting may also cause the high microbial contamination in produce. The efficiency

of MB washing varied from 78 to 99.6% and presented better microbial reduction comparing to

tap water wash at 56 to 95.6%, respectively.

Artificially inoculation acquired the initial load of Escherichia coli (E. coli) and Salmonella

on coriander and Marsh mint by 2.5 to 3.2 and 4.2 to 4.6 log CFU/ml. The reductions of E. coli

and Salmonella on coriander after MB washing are shown in Figure 4A and 4B. Populations of E.

coli on coriander decreased more than 90.0% or 1 log reduction after washing with MNB water,

as well as tap water. Salmonella on coriander washed with MNB water and tap water was

decreased by 99.5% and 99.0%, respectively.

The effect of MB water and tap water for reducing Marsh mint contaminated with E. coli and

Salmonella were shown in Figure 4C and 4D. E. coli was reduced by 95.3 % reductions after

MB washing, while tap water was by 73%. Results showed better Salmonella reduction by 98 to

99.8 % with MB water and tap water. Different level reduction between these two strains may

result from their ability to attach on produce surface which may affect to less washing out,

particularly E. coli.

The best reduction among the six types of tested vegetables was found in asparagus

inoculated with E. coli. Completely reduction from the initial of 2.2-2.7 resulting in 100 %

reduction (not detect at dilution of 10-1

) after washing with MB and tap water (Fig 5A).

However the effect of washing might not be great compared to Salmonella. Washing with MB

water and tap water assisted to reduce Salmonella populations on asparagus by 74 and 80 %

(Figure 5B). Likewise less reduction when washing okra with MB and tap water, microbial

reduction was 52 to 86%. Great result was shown when using MB as washing water, 100%

Salmonella reduction on okra while using tap water gave 98 %. (Fig 5C and 5D)

The initial population of E. coli and Salmonella on Lemongrass were 2.4 to 2.5 log CFU/ml

and 3.3 to 3.7 log CFU/ml (Fig 6A and 6B). Reduction of Salmonella was 98 % reduction. MB

water washing could be the promising method to reduce E. coli and Salmonella on lemongrass,

as well as 98 % reduction on E. coli and 90% Salmonella on ginger (Fig 6C and 6D).

The advantage of using MNB water as washing agent is the possible on less chemical agent

added. This preliminary study was conducted to investigate the possibility and feasibility of

Micro bubble water washing to replace chemical in order to sanitize fresh produce. The results

revealed that MB water contribute to the promising washing agents on microbial reduction which

may be potentially applied to the industry and food services.

However, it was found that additional factors such as water flow rate and washing time

assisted to reduce the microbial load. Application in industry need to consider all factors

involved in washing produce. Apart from the chemical use, the success of the washing depends

on several factors such as target microorganisms, characteristics of produce surface, attachment

of cells to produce surface, formation of resistant biofilms and internalization of microorganisms

(Allende et al., 2008).

98

Expeiment II: Effect of combined microbubbles and various sanitizer agents to reduce

Escherichia coli on Chinese kale (Klintham et al., 2013)

E. coli TISTR780 and Chinese kale was chosen as the model in this study. The final load on

Chinese kale was 5-6 log CFU/mg. The ratio of 200g of Chinese kale per 400 ml of culture

suspension was mixed in PE plastic bag and then let it soaked for 15 min, before let them dry in

laminar flow cabinet for 20 min. Chinese kale samples were MB treated and the samples were

taken for E. coli analysis. Washing condition was done as; 100g of Chinese kale to wash water

10000 mL, with washing time 5 min under continuous generated the microbubbles.

The combination treatments between generated microbubbles and various concentration of

NaOCl (50 ppm and 100 ppm), Acetic acid and Citric acid (0.5% and 1.0% w/v) in washing

contaminated kale showed remarkably the efficiency of sanitizer to eliminate E. coli on Chinese

kale compared to normal washing by tap water. Using the microbubbles water without or with

chemical added have no significantly affect microbial reduction, especially E. coli from Chinese

kale, however, increasing the concentration of sanitizers (NaOCl, Acetic acid and Citric acid) in

washing process contributed to be better microbial reduction (Fig. 7).

Recent experiment: Determination on the efficacy of fine bubbles combined with sanitizers

on reducing Salmonella Typhimurium and Escherichia coli on Thai Mint and Sweet Basil

Recent study by Klintham et al. 2014 (unpublished) was performed to determine the effect of

washing by using the fine bubbles combined with two types of sanitizers on reducing Salmonella

Typhimurium and Escherichia coli on Thai Mint and Sweet Basil. Acidic electrolyte water (20

and 40 ppm free chlorine, AEO), and chlorine dioxide (3 and 5 ppm ClO2) combined with batch

bubble generated by IDEC (Model UFBGALF™) were investigated. The condition of washing

was at 5 min with shaking (60 rpm at 25±2°C).

Determination of oxidation-reduction potential (ORP) in AEO at 20 and 40 ppm AEO were

1090-1200 mV (pH 2.9-3.0) and 1110-1260 mV (pH 2.7-2.9), respectively, which presented

active oxidation, while chlorine dioxide has less ORP value, as 750-850 mV at 3 ppm (pH 5.8),

and 810-890 mV at 5 ppm (pH 5.1). After washing ORP value were decreased although the pH

of wash water were slightly increased. Using 40 ppm AEO with fine bubble treatment

effectively reduced Salmonellae cells on sweet basil and Thai mint with 90.7% and 99.5% log

reduction respectively (P≤0.05). Compared to the results on microbial reduction by aqueous

ClO2, when combined with fine bubbles, washing treatments could reduce Salmonellae on sweet

basil and Thai mint with 99.0% and 90.3% log reduction respectively. Noticeably, E. coli

washing with AEO was reduced to less than 74 % and 86 %, similar to the effect of 5 ppm ClO2

on E. coli reduction (9% and 16%). None of microbes was observed in wash water, which

showed that adding these two sanitizers in bubble water resulting in strong antimicrobial effects

on planktonic bacteria. Although tap water with and without FB also showed microbial

reduction, but there are still viable cells 5-6 log10 CFU/ml found in wash water .

99

CONCLUSIONS

Fine bubble technology provides the opportunities to apply in agriculture and food

products. Using of the fine bubbles technology has unclear mechanism on pesticides degradation,

but Japanese laboratory showed remarkably pesticides reduction when combining fine bubbles

with oxidizing agents like ozone. Recently Thai research group applied combination of fine

bubbles with chemicals like chlorine compounds, organic acids, and others oxidizing agents.

Results showed this technology assist to reduce the pathogenic microorganisms in fresh

vegetables. More studies need to be done to further clarify the antimicrobial mechanism, and to

control possible factors to maximize the utilization of this technology. This technology has the

potential to decrease chemical use, and may be considered as the friendly environmental

technology. Lastly, the challenge of the fine bubbles technology is to find appropriate use, and

to apply to other food process.

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103

Fig. 1. The residual percentage of fenitrothion (FT) in lettuce (A), cherry tomatoes (B) and strawberries (C) at 5 and 10 min after the immersion into the solutions of ozone micro bubble (OMB) treatments by using the gas–water circulation type and the decompression type. Modified from: Ikeura et al., 2011a.

104

Fig. 2. The residual fenithothion (FT) on lettuce, cherry tomatoes and strawberry treated with the

Ozone microbubbles (OMCB) and Ozone millibubbles (OMLB) solutions. Modified

from: Ikeura et al., 2011b.

105

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0 min (Before wash) 15 min (After wash)

Fig. 3. Populations of total microbial count on un-inoculum coriander (3A), Marsh mint (3B),

asparagus (3C), okra (3D), lemongrass (3E), and ginger (3F) before (0 min) and after (15 min) washing by tap water and MNB water.

Ref: Mahakarnchanakul et al., 2010.

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

To

tal m

icro

bia

l (lo

g C

FU

/ml)

92.3% 99.6%

To

tal m

icro

bia

l (lo

g C

FU

/ml)

3C Asparagus 3D Okra

89 %

98% 95.4%

3E Lemongrass

To

tal m

icro

bia

l (lo

g C

FU

/ml)

94.4% 93.4% 97%

3F Ginger

56.5% 87.6%

3A Coriander

95.6% 78.5%

3B Marsh mint

106

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water


Fig. 4. Populations of E. coli on coriander (4A), Marsh mint (4C), and population of Salmonella

on coriander (4B), Marsh mint (4D) before (0 min) and after (15 min) washing by tap

water and MNB water.


E. coli (lo

g C

FU

/ml)

4A Coriander

4C Marsh mint

E. coli (lo

g C

FU

/ml)

95% 90%

Salm

onella

(lo

g C

FU

/ml)

4B Coriander

Salm

onella

(lo

g C

FU

/ml)

99.1% 99.5%

4D Marsh mint

98.2%

107

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water


Fig. 5. Populations of E. coli on asparagus (5A), okra (5C), and population of Salmonella on

asparagus (5B), okra (5D) before (0 min) and after (15 min) washing by tap water and

MNB water.

Ref: Mahakarnchanakul et al., 2010

E. coli (lo

g C

FU

/ml)

100% 100%

Salm

onella

(lo

g C

FU

/ml)

74% 80.6%

E. coli (lo

g C

FU

/ml)

5C Okra

51.6% 86.0%

Salm

onella

(lo

g C

FU

/ml)

5D Okra

98.3%

100%

5A Asparagus 5B Asparagus

108

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water

0

1

2

3

4

5

6

7

8

Tap water MNB water


Fig. 6. Populations of E. coli on asparagus (6A), okra (6C), and population of Salmonella on

asparagus (6B), okra (6D) before (0 min) and after (15 min) washing by tap water and

MNB water.


E. coli (lo

g C

FU

/ml)

100% 100% Salm

onella

(lo

g C

FU

/ml)

74.% 80.6%

E. coli (lo

g C

FU

/ml)

6C Okra

51.6% 86.0%

Salm

onella

(lo

g C

FU

/ml)

6D Okra

98.3%

100%

6A Asparagus 6B Asparagus

109

Fig. 7. Survivor of E.coli on Chinese kale after treatments.

Ref: Klintham et al., 2013.

110

Session III : Analytical approaches for agrochemical determinations

111

APPLICATION OF NON-TARGET ANALYSIS BY HIGH-RESOLUTION MASS

SPECTROMETRY

Chung-Hung Wang1, Yu-Tsung Lee

1, Pei-Ling Huang

1, Yan-Hwa Chu

1

1Product and Process Research Center,

Food Industry Research and Development Institute, Hsinchu, Taiwan


ABSTRACT

High-resolution mass spectrometry (HRMS)-based non-targeted metabolomic analysis is a new

technology used for unbiased collections of food chemical data in food safety inspections

involving unexpected factors. HRMS with high resolving power and high mass spectral accuracy

allows screening of unknown and suspected analytes without reference standards. Its reliable

identification is due to increased selectivity against the food matrix background and thus it

assigns to correct molecular formula of unknown compounds. To introduce the concept and

framework of non-target analysis, we reviewed recent researches involving screening and

identification of chemical contaminant in foods and water by liquid chromatography (LC)-

HRMS. These results indicate that this new detection technique can be used as an early warning

system before running thorough sample analysis for scrutiny.

Keywords: Non-Target Analysis, High-Resolution Mass Spectrometry, Mass Accuracy

112

INTRODUCTION

As the last defense line in food safety evaluation, food inspection and analysis usually target

on specified pesticides, veterinary drugs, and food additives, etc. The problem of these target

specified tests is not being able to detect any chemicals which are not defined in the initial

settings. For examples, incidents of China’s melamine adulterant in milk; illegal dye used in tofu

products and olive oil; phthalate plasticizer added in drinks, jam, jelly, and capsule food in

Taiwan. Nowadays, the international trend is towards to identify emerging food safety issues at

their early stages through non-targeted monitoring analysis technology.

The comparison of the systematic workflows for target and non-target analysis (includes

suspect and unknown screening) is shown in Fig 1. Target analysis is conventional analysis

based on establishing a specified method with standards, i.e. interested analytes, prior to analysis

and monitoring real samples. Compounds are characterized during method development, and

information (retention time, characteristic fragmentation and their ratios) is obtained for further

identification and quantitation of these compounds in real samples. Mass spectrometer with triple

quadrupole (QqQ) technology is useful analyzer for target analysis, due to their high specificity

in multiple reaction-monitoring (MRM) mode and their low limit of detection (LOD). In contrast

to target analysis, non-target analysis starts without any a priori information on the compounds to

be detected and uses general method.

To meet the challenges of analyzing unexpected compounds at low concentrations in

complex matrices, a range of different LC-MS technologies have been bring up in recent years.

Moreover, since sample preparation for non-target analysis is often non-selective and

chromatographic resolution in LC is limited, mass-resolving power (R; defined at full width at

half maximum, FWHM) plays the key role. High resolution mass spectrometry (HRMS) with

high mass accuracy and resolving power has emerged as a powerful tool for non-targeted

analysis. Selectivity in HRMS-based extracted-ion chromatograms is obtained by using narrow

mass-extraction windows (MEWs). About the R of HRMS, EU defines it as 20,000 FWHM

(2002/657/EC, EU/589/2014, SANCO/12571/2013) while World Anti-Doping Agency (WADA)

defines it as 10,000 FWHM. As carrying out an experiment, the resolving power selected

depends on the ratio of the analyte concentration relative to co-eluting matrix interferences and

the sample preparation before LC-MS analysis. With straightforward generic extraction

procedures, typically used in broad screening methods, a first evaluation shows that a resolving

power of 7,000–10,000 can be sufficient for detection of analytes in samples of intermediate

complexity, at levels down to 25 ng/g, with mass errors below 5 ppm (Kellmann et al. 2009). For

consistent and reliable mass assignment (<2 ppm) of analytes at low levels in complex matrices,

a high resolving power (≧50,000) was found to be required (Kellmann et al. 2009; Nielen et al.

2007; van der Heeft et al. 2009). Krauss et al. (2010) overviewed commercially available mass

spectrometers and showed that Orbitrap is increasingly applied due to the combination of high R

(up to 100,000), high mass accuracy (<2 ppm), and a sensitivity down to the femtogram range.

Moreover, in Orbitrap instruments different resolving-power settings can be chosen, involving a

trade-off in data-acquisition rate at increased resolving power (e.g., 12 Hz at 17,500 FWHM and

1 Hz at 140,000 FWHM).

A major advantage of LC-HRMS is the possibility of retrospective analysis of full-scan data,

which enables laboratories to search for new contaminants years after data recording. Other

113

advantages of these instruments are the possibility for a posteriori analysis and their usefulness in

identifying unknown compounds. Moreover, full scan HRMS data can also be recorded under

conditions where fragmentation energy is applied, but without involving a previous precursor

selection (e.g., data dependent MS2 (ddMS

2), variable data-independent acquisition (vDIA).

Newer acquisition modes lie somewhere between all-ion fragmentation (AIF) and traditional

unit-mass MS/MS, and, in this “in-between zone”, the measurement involves multiple scan

events in which consecutively smaller m/z ranges (25–100 Da) are isolated and fragmented (e.g.,

vDIA), thereby still covering the entire m/z range of interest. Hernandez et al. (2012) provided a

critical review about the use of HRMS instruments in environmental sciences, outlining the main

characteristics of HRMS and its great potential in that field. Regarding different fields of food

analysis, Kaufmann (2012) discussed the role of HRMS instruments, including screening

techniques and other aspects related to the potential, or limitations, in quantification and

confirmation processes. In this article, we reviewed recent researches involving with screening

and identification of chemical contaminant in foods and water by LC-HRMS to introduce the

concept and framework of non-target analysis.

SUSPECT SCREENING OF CHEMICAL CONTAMINANTS

For non-target analysis, suspects screening is easier than unknown screening. The use of

HRMS for suspect screening benefits from the advantages of using the full-scan operation mode

which can provide high specificity without limiting the number of simultaneously observed

compounds. The suspect ion list just including the molecular formula allows for the calculation

of an exact m/z of the expected ion, which is in turn extracted from the chromatogram. In the

case of positive findings, further confirmatory steps based solely on structure-derived

information can be employed. The information such as fragment, retention time and so on, can

be established by experimental data of standard. Furthermore, an in-house database can be set up

using this information. Once a database has been established, the parameters of the data

processing software for analyte detection need to be optimized in order to obtain a fit-for-

purpose balance between false positives and false negatives reported by the software (Fig. 2).

About the criteria for the acceptable rates of false positives and false negatives for screening

methods, the USDA Food Safety and Inspection Service (FSIS) requires ≤5% and ≤10%,

respectively (Lehotay et al. 2012; Schneider et al. 2015). Nowadays, most researches of suspect

screening using HRMS focused on pesticides, mycotoxins, fungal metabolites, veterinary drug

(Dzuman et al. 2015; Jia et al. 2014; Gomez-Perez et al. 2012; Wang et al. 2012; Lehner et al.

2011) and plant toxins (Mol et al. 2011).

The procedure of data processing in suspect screening is shown in Fig. 3. Criteria for the

confirmation of suspects need to be carefully chosen in order to minimize the possibilities for

both false-positive and false-negative results. The parameters’ values of criteria are strongly

related to the analysis instruments and their characteristics. The accurate mass and retention time

(RT) is common criterion. In most cases, the mass accuracy in full-scan mode, for different

matrices and concentration levels, is lower than 5 ppm. Gomez-Ramos et al. (2013) indicated

that this value is an acceptable mass accuracy threshold for the confirmation of elemental

composition. In order to avoid both false positives and negatives but, at the same time, missing

the compounds present the sample, the RT window was set at ±0.15 min. Typically, with the

column and gradient selected in that study, the RT precision was very high (<0.02 min for most

of the compounds). Satisfactory results were obtained when combining an accurate mass

114

tolerance of 1 mDa or 5 ppm with a ±0.15 min RT window. The obtained rates for pesticide

identification with the selected parameters at different concentration levels were over 95%

(Mezcua et al. 2009).

One of the major limitations of non-target approach is derived from unexpected matrix

effects or a lack of MS parameter optimization which leads to unexpected false negative results.

It is possible to estimate the efficiency of the automatic screening method in identifying the

selected compounds related to the tested matrix. Jia et al. (2014) successfully developed a

simultaneous screening of 333 pesticide and veterinary drug residues in baby food. In order to

increase sample throughput, full MS scan mode was used for screening to distinguish between

negative and positive samples, and ddMS2 (TopN) fragment spectra were acquired in order to

confirm the presence of the suspected analytes. They showed that as the R set as 70,000 FWHM

(17,500 FWHM for dd-MS2), coeluting matrix compounds from the matrix or noisy peaks can be

easily excluded. The identification of the suspected compounds was based on the measurement

of the accurate mass, RT and fragments. The best results were obtained when mass extraction

windows of 2.5 ppm were employed, providing a high selectivity and reduced probability of

false positives. Mol et al. (2012) studied 21 different vegetable and fruit commodities, a

screening database of 556 pesticides for evaluation of false positives, and a test set of 130

pesticides spiked to the commodities for evaluation of false negatives. The criteria based on

accurate mass of one diagnostic ion and RT resulted in too many false positives. Using relative

response thresholds and one second diagnostic ion effectively reduced false positives. The

threshold was set at half the lowest response obtained for any of the studied matrices. With this

value, the number of false positives was lowered from 88 to 14 without affecting the overall

percentage of detected pesticides. The two-ion approach (other adduct ions, M+1 or M+2

isotopes, fragments) was considered most useful in daily practice. The number of false positives

out of 11,676 pesticide/commodity combinations targeted was 36 (0.3 %). The percentage of

false negatives, assessed for 2,730 pesticide/commodity combinations, was 13 %, 3 %, and 1 %

at the 0.01-, 0.05-, and 0.20-mg/kg level, respectively. Lehner et al. (2011) discussed the

identification criteria for a suspects screening for 200 fungal secondary metabolites in food

samples. The criteria included (1) presence of at least two adduct ion species (protonated

molecules or ammonia/sodium adducts, accurate mass tolerance 3 ppm), (2) the response

threshold of the most intense adduct ion species above 10,000 counts, (3) of the most intense ion

species (I) the peak corresponding to the first 13

C isotopologue had to be present (accurate mass

tolerance 5 ppm) and the ratio of the measured intensity of the I+1 ion to the calculated

(theoretically expected) intensity [Int(I+1)meas]/[Int(I+1)calc] had to be 0.65–1.05, (4) criteria

(1) to (3) had to be fulfilled in at least five scans within a period of 25 s. Their results showed

that applying criteria (1) and (2) to standards in pure solvents and standards in matrix maize led

to zero false-negatives in both cases and to 13 (pure solvents) and 37 (matrix maize) false-

positives. Application of criteria (1) to (3) led to only three false-positives but also two (pure

solvent) and 12 (matrix maize) false-negatives. Applying criteria (1) to (4) led to the nine (pure

solvent) and 16 (standards in matrix maize) false-negatives and zero false-positives.

Acceptable method performance criteria should be established for qualitative analysis

purposes to suit the analytical needs for given applications, and empirical method validation

should be conducted to demonstrate the qualitative performance capabilities of the method.

Lehotay et al. (2015) made recommendations for validation of qualitative methods that meet

common needs for monitoring of chemical contaminants in foods. For regulatory applications in

the European Union (EU), single-laboratory validation of MS-based methods (screening and/or

115

identification) involves analysis of a set of ≥20 different samples each of blanks and spikes at the

RL (Reporting Level, fit-for-purpose threshold concentration above which the laboratory reports

the presence of an analyte in the sample) and/or other concentrations of interest.

Unknown screening for chemical contaminants

In contrast to suspects screening, unknown screening analyzes without any priori information

of those detected compounds. Therefore, comprehensive online databases are indispensable to

provide tentative information of structures identity while processing a huge quantity HRMS data.

Here are some useful databases and mass spectral libraries shown in Table 1 (Milman 2015).

ChemSpider, which is a free database provides structural search access to over 32 million

structures from hundreds of data sources, properties and associated information. There are

several databases that can be used for the searching of pesticides. Commercial applications

include the Merck Index (10,000 compounds total, 500 pesticides estimated) and ChemIndex

(77,000 compounds total, 600 pesticides estimated) both from Cambridge Software.

Here we used the strategy of unknown screening to explore the suspected hazards or

unexpected compounds existed in water samples. The HRMS data were processed and analyzed

by Progenesis QI software (Waters ® ). The workflow includes alignment, peak detection,

deconvolution, component intensity comparison and statistics. In order to increase the efficiency

of data processing, we select the potential peaks with criteria which ANOVA p value < 0.05 and

max fold change >2. The results of multivariate statistical analysis (principal components

analysis, PCA) of these selected peaks showed that reverse osmosis (RO) water was different

from others. Then the S-Plot was drawn for extracting the compounds prominent in RO and other

groundwater samples. Y axis indicated the credit and X axis indicated the response. We picked

up the potential compounds with higher credit and response (shown in Fig. 4). The results of the

tentative identification were shown in Table 2. The mass error of candidate is less than 2 ppm

and isotope similarity is higher than 87.11288. The presence of a distinct isotope signal at M+2,

as is the case for Cl-, Br-, and S-containing compounds, is important for the elemental

composition elucidation. Thus, unknown identification is currently biased to these types of

compounds. We also obtained the structure information of each formula from ChemSpider. Next

step, we will find or predict those characteristics of structure, such as fragment ions and

retentions to check the possibility of each candidate. An unknown screening is challenging to

accomplish the methodology and explore the reality of unknown structures. More experiences of

suspect screening supply the more information of unknown screenings.

CONCLUSION

Nowadays, simultaneous trace analysis of hundreds of unexpected compounds from

different classes is required, preferably in just one run. HRMS instruments can adequately

address such issues and generate meaningful structure suggestions of suspected and unknown

compounds present in experiment samples; however, the main drawbacks are as a result of

insufficient prior optimization of the operational parameters during non-target analysis in full-

scan mode and due to software and database shortcomings. The problem comes from the

software which needs to be enhanced for a better fit-for-purpose performance in non-target

analysis and for an increase in data processing speed in quantitation processes. Overcoming this

drawback will improve non-target analysis technology to perform in a routine detection and an

116

early warning detection system. Additionally it could be used to continuously monitor for new

food contaminants as a complement to the specific targeted analysis that is today’s foundation of

food safety analysis.

REFERENCES

Dzuman, Z., M. Zachariasova, Z. Veprikova, M. Godula, and J. Hajslova. 2015. Multi-analyte high performance liquid chromatography coupled to high resolution tandem mass spectrometry method for control of pesticide residues, mycotoxins, and pyrrolizidine alkaloids. Analytica Chimica Acta 863:29-40.

Gomez-Perez, M. L., P. Plaza-Bolanos, R. Romero-Gonzalez, J. L. Martinez-Vidal, and A. Garrido-Frenich. 2012. Comprehensive qualitative and quantitative determination of pesticides and veterinary drugs in honey using liquid chromatography-Orbitrap high resolution mass spectrometry. Journal of Chromatography A 1248:130-138.

Gomez-Ramos, M. M., C. Ferrer, O. Malato, A. Aguera, and A. R. Fernandez-Alba. 2013. Liquid chromatography-high-resolution mass spectrometry for pesticide residue analysis in fruit and vegetables: Screening and quantitative studies. Journal of Chromatography A 1287:24-37.

Hernandez, F., J. V. Sancho, M. Ibanez, E. Abad, T. Portoles, and L. Mattioli. 2012. Current use of high-resolution mass spectrometry in the environmental sciences. Analytical and Bioanalytical Chemistry 403 (5):1251-1264.

Jia, W., X. G. Chu, Y. Ling, J. R. Huang, and J. Chang. 2014. High-throughput screening of pesticide and veterinary drug residues in baby food by liquid chromatography coupled to quadrupole Orbitrap mass spectrometry. Journal of Chromatography A 1347:122-128.

Kaufmann, A. 2012. The current role of high-resolution mass spectrometry in food analysis. Analytical and Bioanalytical Chemistry 403 (5):1233-1249.

Kellmann, M., H. Muenster, P. Zomer, and H. Mol. 2009. Full scan ms in comprehensive qualitative and quantitative residue analysis in food and feed matrices: How much resolving power is required? Journal of the American Society for Mass Spectrometry 20 (8):1464-1476.

Krauss, M., H. Singer, and J. Hollender. 2010. LC-high resolution MS in environmental analysis: from target screening to the identification of unknowns. Analytical and Bioanalytical Chemistry 397 (3):943-951.

Lehner, S. M., N. K. N. Neumann, M. Sulyok, M. Lemmens, R. Krska, and R. Schuhmacher. 2011. Evaluation of LC-high-resolution FT-Orbitrap MS for the quantification of selected mycotoxins and the simultaneous screening of fungal metabolites in food. Food Additives and Contaminants Part a-Chemistry Analysis Control Exposure & Risk Assessment 28 (10):1457-1468.

Lehotay, S. J., A. R. Lightfield, L. Geis-Asteggiante, M. J. Schneider, T. Dutko, C. Ng, L. Bluhm, and K. Mastovska. 2012. Development and validation of a streamlined method designed to detect residues of 62 veterinary drugs in bovine kidney using ultra-high performance liquid chromatography - tandem mass spectrometry. Drug Testing and Analysis 4:75-90.

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Lehotay, S. J., Y. Sapozhnikova, and H. G. J. Mol. 2015. Current issues involving screening and identification of chemical contaminants in foods by mass spectrometry. Trac-Trends in Analytical Chemistry 69:62-75.

Mezcua, M., O. Malato, J. F. Garcia-Reyes, A. Molina-Diaz, and A. R. Fernandez-Alba. 2009. Accurate-mass databases for comprehensive screening of pesticide residues in food by fast liquid chromatography time-of-flight mass spectrometry. Analytical Chemistry 81 (3):913-929.

Milman, B. L. 2015. General principles of identification by mass spectrometry. Trac-Trends in Analytical Chemistry 69:24-33.

Mol, H. G. J., R. C. J. Van Dam, P. Zomer, and P. P. J. Mulder. 2011. Screening of plant toxins in food, feed and botanicals using full-scan high-resolution (Orbitrap) mass spectrometry. Food Additives and Contaminants Part a-Chemistry Analysis Control Exposure & Risk Assessment 28 (10):1405-1423.

Mol, H. G. J., P. Zomer, and M. de Koning. 2012. Qualitative aspects and validation of a screening method for pesticides in vegetables and fruits based on liquid chromatography coupled to full scan high resolution (Orbitrap) mass spectrometry. Analytical and Bioanalytical Chemistry 403 (10):2891-2908.

Nielen, M. W. F., M. C. van Engelen, R. Zuiderent, and R. Ramaker. 2007. Screening and confirmation criteria for hormone residue analysis using liquid chromatography accurate mass time-of-flight, Fourier transform ion cyclotron resonance and orbitrap mass spectrometry techniques. Analytica Chimica Acta 586 (1-2):122-129.

Schneider, M. J., S. J. Lehotay, and A. R. Lightfield. 2015. Validation of a streamlined multiclass, multiresidue method for determination of veterinary drug residues in bovine muscle by liquid chromatography-tandem mass spectrometry. Analytical and Bioanalytical Chemistry 407 (15):4423-4435.

van der Heeft, E., Y. J. C. Bolck, B. Beumer, A. Nijrolder, A. A. M. Stolker, and M. W. F. Nielen. 2009. Full-scan accurate mass selectivity of ultra-performance liquid chromatography combined with time-of-flight and orbitrap mass spectrometry in hormone and veterinary drug residue analysis. Journal of the American Society for Mass Spectrometry 20 (3):451-463.

Wang, J., W. Chow, D. Leung, and J. Chang. 2012. Application of ultrahigh-performance liquid chromatography and electrospray ionization quadrupole orbitrap high-resolution mass spectrometry for determination of 166 pesticides in fruits and vegetables. Journal of Agricultural and Food Chemistry 60 (49):12088-12104.

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Table 1. Useful databases and mass spectral libraries for the identification of the non-target analysis

Name Coverage Information

Chemical and biochemical database

CA Over 154 millions of CAS registry number, references and abstracts,

compounds/substances partly properties and NMR spectra, reactions,

commercially available and regulated chemicals

ChemSpider Over 32 millions of structures Accurate ion mass, elements, structural

descriptors/identifiers,

chemical and physical, properties, spectra,

biomedical information, pharmacological action,

literature references and patents, related databases,

chemical vendors, and so on

PubChem 19 Millions of unique compound Structural descriptors/identifiers, related compounds,

structures (in 2008) biomedical information, bioassay, pharmacological action,

literature references and patents, chemical vendors, and so

on

Mass spectral libraries

NIST, MS2

9,344 Compounds, 234,284

spectra Containing peptides

METLIN 12,057 Compounds, 61,872

spectra Metabolites

MassBank >15,000 Compounds, 40,889

spectra Biocompounds (include MS

1,MS

n)

LipidBlast 119,200 Compounds, 212,516

spectra Lipids (predicted MS

2)

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Table. 2. The compounds prominent in groundwater and RO

Compound RT Formula Adducts

Mass Error

(ppm)

Isotope Similarity

The number of structures

in ChemSpider

for each formula

b

Compounds prominent in groundwater(Hsinchu)

302.1728n a 16.90 C15H26O6

M+H, M+NH4, M+Na, M+H-H2O

-0.58043 95.09865 129

262.2373m/z 7.15 C18D10H8 M+NH4 -0.76413 94.85578 1

C15H31N2+ M+Na -0.57578 97.16693 9

452.3350n 5.11 C23H48O8 M+H, M+Na, M+NH4 0.20794 97.11860 5

C26H49N2PS M+H, M+Na, M+NH4 -0.87312 92.90380 1

C24H44N4O4+2 M+H, M+Na, M+NH4 -0.26516 98.38726 1

498.4002m/z 5.11 C25H52O8 M+NH4 0.34617 98.20752 5

385.3168m/z 4.45 C20H37N3O3 M+NH4 -0.02682 95.46042 3

C20H48O2S2 M+H -0.08002 87.11288 1

290.2685m/z 10.88 C17H35N2+ M+Na -0.78842 96.33369 1

Compounds prominent in RO

546.3240n 4.97 C29H46N4O4S M+NH4, M+Na, M+H 0.01019 90.41096 3

502.2981n 4.74 C26H48O5P2 M+NH4, M+Na, M+H 0.82518 96.71692 1

C27H42N4O3S M+NH4, M+Na, M+H 0.70373 90.10872 14

C35H38N2O M+NH4, M+Na, M+H -0.57240 88.23453 7

590.3499n 5.15 C31H50N4O5S M+NH4, M+Na, M+H -0.61506 90.06301 1

C30H56O7P2 M+NH4, M+Na, M+H -0.40840 96.69614 1

120

678.4021n 5.59 C35H58N4O7S M+NH4, M+Na, M+H -0.78947 89.65370 3

634.3761n 5.37 C32H60O8P2 M+NH4, M+Na, M+H -0.35113 96.97243 1

C33H54N4O6S M+NH4, M+Na, M+H -0.44729 90.29635 9

458.2719n 4.56 C25H38N4O2S M+NH4, M+Na, M+H 0.82041 90.22029 65

C27H40NO3S+ M+NH4, M+Na, M+H -0.91218 89.03847 1

C29H37N3P+ M+NH4, M+Na, M+H -0.11353 91.83047 1

C33H34N2 M+NH4, M+Na, M+H -0.57832 88.32950 10

282.1673n 3.83 C11H20N7O2+ M+H, M+Na, M+NH4 0.00113 98.52783 2

238.1412n 3.57 C14D10H2O3 M+H, M+Na, M+NH4 -0.96292 95.01603 1

326.1935n 4.01 C21H28NS+ M+NH4, M+Na, M+H -0.69436 87.87890 2

C13H24N7O3+ M+NH4, M+Na, M+H -0.13335 98.03543 3

370.2197n 4.20 C23H32NOS+ M+NH4, M+Na, M+H -0.47734 88.60777 4

a "n" means "neutral mass"

b The criteria of the accurate mass tolerance is 2 ppm

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Fig. 1. Comparison of systematic workflows for target and non-target analysis in detecting samples

122

Fig. 2. Proposed workflow for identification strategies of non-target analysis

123

Fig. 3. The procedure of data processing in suspect screening

124

Fig. 4. Statistics analysis of S-Plot for extracting the compounds prominent in groundwater and RO.

125

CHEMOMETRIC APPROACH TO THE OPTIMIZATION OF HS-

SPME/GC-MS FOR THE DETERMINATION OF MULTICLASS

PESTICIDE RESIDUES IN FRUITS AND VEGETABLES

Lukman B. Abdulra’uf * and Guan H. Tan

†

Department of Chemistry, Faculty of Science,

University of Malaya, Pantai Valley, Kuala Lumpur, 50603, Malaysia. *[email protected],

†[email protected] (+60379674193)

ABSTRACT

A Headspace Solid Phase Microextraction (HS-SPME) method was developed using multivariate

experimental designs, which was conducted in two stages. The significance of each factor was

estimated using the Plackett-Burman (P-B) design, for the identification of significant factors,

followed by the optimization of the significant factors using central composite design (CCD). The

multivariate experiment involved the use of Minitab® statistical software for the generation of a

27-4

P-B design and CCD matrices. The method performance evaluated with internal standard

calibration method produced good analytical figures of merit with linearity ranging from 1 – 500

µg/kg with correlation coefficient greater than 0.99; limit of detection (LOD) and limit of

quantification (LOQ) were found between 0.35 and 8.33 µg/kg and 1.15 and 27.76 µg/kg,

respectively. The average recovery was between 73 % and 118 % with relative standard

deviation (RSD = 1.5 – 14 %) for all the investigated pesticides. The multivariate method helps

to reduce sampling time and improve analytical throughput.

Kewords: GC-MS, SPME, multivariate design, central composite design, pesticide residues,

fruits and vegetables

INTRODUCTION

An estimate of 1 billion people went hungry in 2010, and with the ever increasing world

population, there is need for 70% increase in global food production by the year 2050.1 The

increase in world population which has led to drastic increase in demand for food supply has also

led to immeasurable rise in the application of chemical pesticides and fertilizers.2 To increase

agricultural production and meet the growing demand for food, pesticides are used for control of

pest and vector of plant diseases.3 Pesticides are also used in non-agricultural activities to control

and eradicate carriers of vector borne diseases, such as malaria, yellow fever, typhoid fever and

dengue, which are major public health concerns.4-8

The production and applications of pesticides

in agriculture and non-agricultural purposes have led to steady increase in food production, high

food quality and reduced incidence of illness due to insect-borne diseases. However, their

126

continuous use has negative impact on the environment and their presence in soil, air, water and

food poses a potential health risk due to their biocide activity.2, 3, 9-12

Therefore, pesticides must

be used efficiently and effectively in order to strike a balance between their expected benefits

and the possible risk to human health. This will enable their economic viability and

environmental sustainability.13

Although the use of pesticides has helped to increase food production, there is need to strike

balance between the expected benefit and the possible risks to human health.3 Therefore, there is

an urgent need for quality control monitoring of the use of such pesticides on fruits and

vegetables for safety purposes.14

Analytical study is undertaken in order to obtain information on

the quality and quantity of contaminants present in the sample. The extraction and subsequent

analysis of pesticide residues and other contaminants in fruit and vegetable samples is becoming

increasingly important due to the health hazards caused by their accumulation in human tissues.15

Solid Phase Microextraction (SPME) is a very attractive extraction technique in sample

preparation that results in high throughput analysis, and remarkable analytical characteristics,

including linearity, reproducibility, repeatability, low and improved limits of detection and

quantitation, high selectivity, sensitivity and versatility with minimum matrix interferences.16-19

It combines sampling, isolation, concentration and enrichment and sample introduction into

analytical instruments in a single and uninterrupted sampling step, which results in high

throughput analysis.16, 19-21

SPME was developed to overcome the problems associated with the

solvent-based, time consuming conventional techniques, which are multistep and usually require

a large amount of samples and solvents that can cause environmental pollution and be hazardous

to human health.

There are different parameters that influence the partitioning of the analytes between the

sample matrix and the SPME fiber. When considering the optimization of parameters for fruits

and vegetables analysis, the complex nature of the sample should be taken into consideration.22

The amount of analytes extracted from fruit and vegetable samples depend on the nature of the

stationary phase (fiber) and on the properties of the sample matrix. The most important method

used in the optimization of extraction parameters is the consideration of the thermodynamic and

theoretical models,23

in the selection of a particular procedure for the development of method for

the determination of pesticides in fruits and vegetable samples.

The univariate optimization of SPME parameters involves optimizing each factor one at a

time, in which other factors are kept constant except for the one being optimized and it involves

many experiments.24

This does not allow the estimation of possible interaction between the

studied factors. Multivariate experimental design helps to identify the significant factors that

maximize the response of an experiment. It also helps to improve the yield or chromatographic

separation by optimizing the significant factors using response surface methodology or central

composite design (CCD). It saves time and requires few experimental runs and can be used for

quantitative modeling of mathematical relationships between factors and response.25, 26

Its use is

aimed to understand the effect of each factor and model the relationship between the factors and

response with a minimal number of experiments carried out in an orderly and efficient manner.27

In this study, a fast and robust Headspace (HS)-SPME-gas chromatography-mass

spectrometry (GC-MS) method was developed for the simultaneous determination of fourteen

multiclass pesticide residues in fruit and vegetable samples using Plackett-Burman (P-B) design

to study the significance of various factors and its optimization using CCD.

127

MATERIALS AND METHODS

Reagents and Solutions

Pesticide standards (fenobucarb, ethoprop, diazinon, chlorothalonil, fenitrothion, methyl

parathion, chlorpyrifos, thiobencarb, quinalphos, endosulfan I, endosulfan II, bifenthrin,

fenpropathrin and permethrin) at 100 µg/mL and 1-chloro-3-nitrolbenzene (1000 µg/mL) used as

internal standard with more than 95 % purity, were purchased from AccuStandard Inc. New

Haven CT, U.S.A. A working standard solution containing the pesticides was prepared daily by

diluting the stock solution in methanol to a concentration of 10µg/mL, and kept at 4 0C before

use. All solvents used were pesticide grade: methanol, acetone and acetonitrile were purchased

from Fisher Scientific, Loughborough, U.K. Sodium chloride, sodium sulphate, ammonium

chloride were purchased from Merck. The pH buffer solutions 4, 6, 8 – 10 and 5 – 7 were

purchased from Fisher Scientific and Sigma-Aldrich respectively. Millipore filtered (0.45 µm)

deionized water was used for method development.

Sample Preparation

For SPME method development, 100 g of pesticide free fruits and vegetables were accurately

weighed, finely chopped and homogenized in a blender. A known aliquot of the homogenized

sample was then weighed into a separate 20 mL amber glass vial containing the internal standard

and diluted accurately with Milli-Q filtered deionized water containing 10 % of NaCl to make up

a total mass of 5 g. The mixture was then spiked with a known amount of the working standard

solution to prepare a concentration of 50µg/kg. The mixture was then homogenized at 3000 rpm

for 5 min and subjected to SPME procedure. Fruit and vegetable samples used for method

development, calibration and recovery studies were first analyzed to ensure the absence of the

target pesticide residues.28-31

HS-SPME Procedure

The SPME fibers (Supleco, Bellefonte, PA USA) were conditioned in the GC/MS injection at

250 0C for 30 min (PDMS and PDMS/DVB) and 280

0C for 1 hr (PA), prior to their first use as

recommended by the manufacturer. Optimization of parameters and analysis were performed in a

20 mL amber glass vial containing 5 mL of Milli-Q filtered deionized water containing 10 % of

NaCl and spiked with 50 µL of the working standard solution to give a concentration of

0.1µg/mL. The vial containing the sample was shaken ultrasonically for 5 min, agitated and

incubated for 5 min at 60 0C in the autosampler agitator, followed by the exposure of the fiber to

the headspace of the sample in the vial sealed with PTFE/silicone septum.

GC Condition

The extraction and analysis of pesticides were carried out with CTC CombiPAL autosampler

equipped with agitator and needle heater (for fiber conditioning and inter-extraction clean up)

coupled to a GC-MS (Shimadzu QP2010Series) and operated in the split/splitless mode at an

injection temperature of 270 0C. The separation of target analytes was achieved on a DB-5MS

fused capillary column containing 5 % diphenyl and 95 % dimethylpolysiloxane (30 m x 0.25

128

mm i.d. x 0.25 µm film thickness). The injection port of the GC was equipped with a high-

pressure Merlin Microseal septumless injection kit and a silanized narrow bore liner (78.5 x 6.5

mm o.d x 0.75 mm i.d). Helium (carrier gas) was set to a constant flow rate of 1.3 mL/min with

linear velocity of 42 cm/sec. The GC column oven temperature program was set as follows.

Initially set at 60 0C for 2 min, ramped at 30

0C/min to 180

0C, then ramped to 210

0C at 5

0C/min,

and finally to 270 0C held for 5 min, for a total runtime of 24.50 min. The MS operation

condition includes transfer line of 300 0C, ion source of 200

0C, electron ionization (EI) of 70 eV.

The optimization of methods was done in scan mode while quantitation was done in selected ion

monitoring (SIM) mode. A target ion (most abundance ion) and two other reference ions were

monitored for the target analytes. The investigated pesticides were identified by comparing the

mass spectrum obtained for each analyte to that of the reference compound in GC-MS library

using the US National Institute of Standard and Technology (NIST) and PESTANA libraries

search. In case of co-elution, easy spectral identification and integration was achieved by using

the deconvolution feature of the GC-MS system. The P-B and the CCD matrices were performed

and estimated with Minitab® statistical software package version 16 (Minitab Inc., State College,

USA).

RESULTS AND DISCUSSION

Several parameters affecting the efficiency of SPME of pesticide residues in fruits and

vegetables were optimized. The optimization involved a two-step design: the screening design

was used to determine the significant factors and the optimization design for estimating the best

experimental conditions

P-B Design

The P-B design matrix with a 27–4

(resolution III) reduced factorial was generated for the

screening of the most important factors affecting the SPME efficiency and recovery of pesticide

residues from fruit and vegetable samples. It contains experimental runs of a multiple of four (4,

8 , 12, 16, etc.) and the factors are one less than the number of experiments.26, 32

It helps for the

estimation of the significant factors affecting extraction efficiency. It does not yield the exact

quantity, but provides valuable information on each variable with relatively few and reasonable

experimental runs.32, 33

The factors and level of variables selected for P-D design are shown in

Table 1. A 27–4

P-B design matrix (Table 2), was used to run the experiment for the

determination of main effects of the factors under investigation. The P-B design consists of 12

runs, conducted in duplicate, to annul the effects of extraneous variables. 34

The main effect of each factor was estimated using least square regression which indicates

the significance in relation to the response (TCPA). In the Pareto chart (Fig 1a), the length of the

bar is proportional to the absolute value of the main effect 32-34

, while the vertical line indicates

95 % confidence level. The normal plot (Fig 1b) shows the significance of each factor (estimated

using ANOVA test) and the magnitude of various effects, while the residual plots (Fig 1c) show

that the measurement deviation is randomly distributed around the mean. The main effect plot

(Fig 1d), as indicated by the slope of the plots, shows that when extraction temperature and

extraction time increase from low value to high value, the extraction efficiency also increases

with decrease in stirring rate and pH, while other factors such as salt addition, desorption time

129

and desorption temperature show no significant effect. The extraction temperature is the most

important factor followed by the extraction time. As can be observed from the normal plot (Fig

1b) extraction temperature and time shows positive effects, while pH and stirring rate show

negative effect. Therefore, for the optimization step, all other factors were fixed, while extraction

temperature, time, pH and stirring rate were considered for further optimization.

CCD

The screening experiment obtained by the use of P-B design indicates thatdesorption time,

desorption temperature and salt addition do not affect extraction efficiency to a significant extent.

Therefore, they were fixed according to the optimal value estimated using the univariate

experiments (desorption time, 7 min; desorption temperature, 270 0C; salt addition, 10 %) (Data

not shown). The extraction time, extraction temperature, salt addition and pH, which are the

significant variables, were further optimized by the use of second-order CCD utilizing a response

surface methodology (RSM). The number of points in CCD contains a factorial run of 2k, axial

runs of 2k and Co center point runs. Therefore, the total experimental runs (N) of CCD is given

by: N = 2k + 2k + Co, where k and Co are the number of variables and the number of center points,

respectively.34, 35

In order to reduce the effect of uncontrolled variables, the CCD experiments

were run in a random manner. The CCD design includes 16 cube points, 7 center points in cube,

8 axial points and 0 center point in axial with α = 2 (selected to establish rotatability conditions)

and a total of 31 randomized runs. The significant variables involved in the generation of CCD,

their levels and the design matrix are shown in Table 3.

The total chromatographic peak area (TCPA) corresponding to the 14 investigated pesticides

for the experimental runs presented in Table 3, were used to obtain the response surface plot as

shown in Fig 2. The desirability function was first fixed by assigning values of 0.0 (undesirable),

0.5 (medium desirability) and 1.0 (very desirable). The global desirability surface response in 3D

plot was obtained for the optimized parameters as shown in Fig 2. The second order response is

utilized because it has flexibility, the ability to give an approximation of the true value and the

parameters to be easily estimated 36

.

The surface plot (Fig 2) and response optimizer plot (Fig 3) were used to indicate the optimal

conditions, and it was observed that the overall response desirability of the independent variables

in the experimental domain was obtained at extraction temperature greater than or equal to 62 0C,

while optimum extraction time can be for 34 min or longer, the optimum stirring rate was at 351

rpm or lower and the optimum pH was greater than or equal to 6. The result is in good agreement

with the P-B design as represented by the main effect plot (Fig 5.24) where extraction efficiency

is increased by increasing of extraction temperature and time while it is increased by decreasing

of stirring rate and pH value. Consequently, the optimal extraction conditions are as follows:

Temperature, 65 0C; time, 35 min; salt addition, 10 %; stirring rate, 350 rpm; pH, 6; desorption

time, 7 min; desorption temperature, 270 0C. The chromatogram of the 14 investigated pesticides

spiked in water sample and analyzed under the optimized conditions is shown in Fig 4. The

chromatogram was integrated for each peak and the ion fragmentations obtained were compared

with the NIST library (See Supplementary Information Fig S1-S15).

130

Validation of Analytical Figures of Merit

The validation of analytical methodology has been observed to be a quality assurance step in

method development,37

used to confirm the method performance and its suitability for the

intended purpose. In the present study, the figures of merit of analytical methodology of the

developed method was validated in terms of linearity, accuracy, intra- and inter-day precisions,

limits of detection (LOD) and quantification (LOQ) using the optimized HS-SPME parameters.

Although, validation of figures of merit of analytical methodology has been described to be a

time consuming activity, it is very essential in order to ensure optimal utilization of analytical

resources. 38, 39

The linearity was estimated using set of calibration curves prepared with concentrations

ranging from 1 – 500 µg/kg, with an internal standard calibration method. The peak area ratio

which is the ratio of the peak area of analytes to the peak area of internal standard was plotted

against the concentration of analytes. As shown in Table 4, the calibration curves were linear

over the tested concentration range and the correlation coefficients (r2) were greater than 0.99 for

all the investigated pesticides. Table 5 shows the precisions and accuracies (relative recoveries)

of the developed method in fruit and vegetable samples. The intra-day precision varies from 1.56

to 13.9 % while the intermediate precision varies from 2.4 to 14.9 %. The relative recoveries of

the spiked fruit and vegetable samples range from 73.3 to 118.5 % which were acceptable

according to the SANCO guideline.40

The recoveries obtained in vegetable samples were slightly higher than those obtained in

fruit samples, this could be attributed to the presence of suspended solid particles and high

molecular mass substances such as pectin and sugar present in the fruit samples,41-43

although

matrix interference were completely eliminated by appropriate dilution of the samples. It was

also observed that, better recoveries and precisions were achieved at higher spiked levels. All the

parameters validated in this study were based on the method validation requirements of the

European Union.40

The LOQ (S/N =10) and LOD (S/N=3) values obtained (Table 6) are in most

cases below the first calibration level and are lower than the maximum residue levels (MRL)

allowed by Codex Alimentarius and the European Union.44

The LOD range from 0.35 to 8.33

µg/kg, while the LOQ was between 1.15 and 27.76 µg/kg.

The HS-SPME method developed in this study was subsequently applied to the analysis of

apple, tomato, cucumber and cabbage samples purchased from a local Malaysian wet market.

The real sample analysis was conducted in order to further verify the reliability and robustness of

the developed method. A total of 220 samples were analyzed, and three samples each of tomato

and cabbage were found to contain chlorothalonil, while one sample of tomato contains

permethrin. One sample of apple was also found to contain chlorpyrifos, with three samples of

cabbage found to contain chlorothalonil. However, all fruits and vegetables found to contain the

target pesticides were far below the maximum residue levels allowed by the European Union

and the Codex Alimentarius Commission.44

All other pesticides investigated in the selected

commodities were either not detected or were detected below the limits of quantification and

thus were not quantified.

The use of chemometric approach to the screening and subsequent optimization of extraction

parameters has helped to reduce analysis time and also helped to determine the best optimized

parameters. The combination of microextraction and chemometrics, as can be observed in this

study, enhances better recoveries and precisions and also improves detectability of the target

analytes as an improved method validation. Further studies should be focused on the use of sol-

131

gel prepared, ionic liquid, supramolecular molecules and molecularly imprinted polymer

coatings as the extraction phase to increase the range of analytes that can be qualitatively and

quantitatively analyzed in a wide range of environmental samples.

AUTHOR INFORMATION

Corresponding Author

*Phone: +60379674247. Fax +6079674193. E mail: [email protected]

Funding Sources

Financial support of the University of Malaya Center for Research Management through the

IPPP grant (PV09/2011A) and UMRG grant (RG227/12AFR)

Notes

The authors declare no competing financial interest.

ACKNOWLEDGMENT

The University of Malaya Research Management Center is gratefully acknowledged for the

permission given to publish this research work.

ABBREVIATIONS USED

SPME, solid phase microextraction; HS, headspace; PDMS, polydimethylsiloxane,; DVB,

divinylbenzene; PA, polyacrylate; GC-MS’ gas chromatography-mass spectrometry; TCPA, total

chromatographic peak area; ANOVA, analysis of variance; CCD’ central composite design;

RSM, response surface methodology; S/N, signal to noise ratio; SIM, selective ion monitoring;

RSD, relative standard deviation

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135

Table 1. Factors and levels of variables

Variables Levels

Low (-) High (+)

Extraction temperature (0C) 30 60

Extraction time (min) 30 60

Salt addition (%, v/v) 5 10

Stirring rate (rpm) 300 600

pH 4 8

Desorption time (min) 5 10

Desorption temperature (0C) 250 270

Table 2. Plackett-Burman design matrixa

Ext. temp

(0C)

Ext. time

(min)

Salt add

(%)

Stirring rate

(rpm)

pH Des. time

(0C)

Des. temp

(0C)

60 30 5 600 4 10 270

60 60 10 600 8 10 270

30 60 5 600 8 10 250

60 30 10 600 8 5 250

60 30 5 300 8 5 250

30 30 5 600 4 5 270

60 60 5 300 4 10 250

30 30 5 600 4 5 270

60 30 5 600 4 10 270

60 60 10 300 4 5 270

30 30 10 300 8 10 270

60 30 10 600 8 5 250

30 30 10 300 4 10 250

30 60 5 300 8 5 270

30 30 10 300 4 10 250

30 60 5 300 8 5 270

60 60 10 600 8 10 270

30 60 10 600 4 5 250

30 60 5 600 8 10 250

60 60 5 300 4 10 250

30 30 10 300 8 10 270

60 30 5 300 8 5 250

30 60 10 600 4 5 250

60 60 10 300 4 5 270 a Generated using Minitab® statistical software

136

Table 3. Factors, levels and CCD design matrix

Variables Level Star points (α=2)

Low (–) Central (0) High (+) –α +α

Extraction temp. (0C) 30 45 60 15 75

Extraction time (min) 30 45 60 15 75

pH 4 6 8 2 10

Stirring rate (rpm) 300 450 600 150 750

StdOrder RunOrder PtType Blocks Temp Time Stirring pH

10 1 1 1 60 30 300 8

6 2 1 1 60 30 600 4

8 3 1 1 60 60 600 4

13 4 1 1 30 30 600 8

18 5 -1 1 75 45 450 6

30 6 0 1 45 45 450 6

26 7 0 1 45 45 450 6

11 8 1 1 30 60 300 8

14 9 1 1 60 30 600 8

29 10 0 1 45 45 450 6

15 11 1 1 30 60 600 8

20 12 -1 1 45 75 450 6

24 13 -1 1 45 45 450 10

5 14 1 1 30 30 600 4

27 15 0 1 45 45 450 6

12 16 1 1 60 60 300 8

31 17 0 1 45 45 450 6

2 18 1 1 60 30 300 4

3 19 1 1 30 60 300 4

7 20 1 1 30 60 600 4

22 21 -1 1 45 45 750 6

9 22 1 1 30 30 300 8

25 23 0 1 45 45 450 6

4 24 1 1 60 60 300 4

16 25 1 1 60 60 600 8

17 26 -1 1 15 45 450 6

19 27 -1 1 45 15 450 6

23 28 -1 1 45 45 450 2

21 29 -1 1 45 45 150 6

28 30 0 1 45 45 450 6

1 31 1 1 30 30 300 4 a Generated using Minitab® statistical software

137

Table 4. Linearity range (µg/kg) of the developed HS-SPME method

Pesticides Range Apple Tomato Cucumber Cabbage

(µg/kg) r2 r

2 r

2 r

2

Fenobucarb 2.5 – 500 0.9975 0.9996 0.9985 0.9976

Ethoprophos 2.5 – 250 0.9981 0.9986 0.9975 0.9979

Diazinone 2.5 – 250 0.9987 0.9948 0.9981 0.9980

Chlorothalonil 10 – 500 0.9987 0.9975 0.9978 0.9989

Parathion-methyl 1 – 250 0.9986 0.9994 0.9988 0.9964

Fenitrothion 2.5 – 200 0.9989 0.9995 0.9983 0.9952

Chlorpyrifos 5 – 500 0.9980 0.9979 0.9981 0.9985

Thiobencarb 5 – 250 0.9982 0.9950 0.9984 0.9977

Quinalphos 2.5 – 125 0.9985 0.9991 0.9981 0.9968

Endosulfan I 5 – 250 0.9980 0.9967 0.9990 0.9976

Endosulfan II 10 – 250 0.9988 0.9992 0.9978 0.9987

Bifenthrin 1 – 500 0.9985 0.9989 0.9983 0.9982

Fenpropathrin 1 – 50 0.9976 0.9938 0.9984 0.9978

Permethrin 5 – 100 0.9969 0.9976 0.9989 0.9973

138

Table 5. Accuracy, intra- and inter-day precisions of the pesticides in fruit and vegetable samples

Pesticides Added

(µg/kg)

Apple Tomato Cucumber Cabbage

Intra

(%)

Inter

(%)

Accuracy

(%)

Intra

(%)

Inter

(%)

Accuracy

(%)

Intra

(%)

Inter

(%)

Accuracy

(%)

Intra

(%)

Inter

(%)

Accuracy

(%)

Fenobucarb 50

100

150

11.5

8.2

2.74

12.9

9.0

4.0

80.9

96.2

103.5

2.8

2.2

2.2

6.0

2.4

3.1

105.0

75.6

95.5

7.4

5.1

3.1

10.4

6.1

4.4

76.40

80.70

89.19

9.1

4.9

4.9

12.6

6.1

6.8

76.8

77.4

92.6

Ethoprophos 50

100

150

13.6

8.3

4.3

14.9

8.9

4.5

79.1

95.0

102.4

13.2

3.0

3.2

14.5

3.3

5.0

80.0

75.4

91.6

8.9

3.0

5.3

9.6

3.5

5.3

83.11

86.89

88.59

12.2

7.1

5.1

14.3

8.0

5.7

89.2

81.8

89.4

Diazinon 50

100

150

5.5

5.1

7.4

9.2

6.0

6.3

77.8

88.7

101.9

10.4

3.7

3.2

13.7

5.6

6.0

82.3

75.5

102.8

8.4

5.0

2.8

12.4

6.4

3.2

88.94

91.72

103.85

9.9

9.0

8.3

10.8

9.3

8.7

85.8

87.6

106.1

Chlorothalonil 50

100

150

11.5

5.1

6.1

12.4

9.8

7.0

76.2

81.0

104.2

8.8

7.1

3.8

10.2

7.6

4.7

112.0

109.3

115.0

11.4

2.7

4.2

13.2

4.5

4.5

81.86

105.11

117.0

12.9

9.0

7.8

14.8

10.2

8.7

113.8

113.6

90.5

P. Methyl 20

50

100

6.8

3.2

4.3

10.7

10.6

6.4

73.3

80.4

98.0

10.5

6.1

4.4

11.0

6.7

4.7

80.3

96.9

105.0

10.7

8.6

4.4

12.0

9.4

4.8

77.66

78.43

78.56

12.5

10.5

8.8

14.1

12.0

10.0

76.0

78.4

103.3

Fenitrothion 50

100

150

5.6

4.4

4.3

6.6

7.0

4.3

102.4

106.4

107.6

12.1

4.0

5.5

13.6

6.4

7.4

85.5

99.8

107.6

10.8

4.6

3.7

11.4

5.3

4.2

90.71

109.55

93.06

12.7

9.3

5.6

14.4

10.8

6.4

117.9

106.6

105.1

Chlorpyrifos 50

100

150

7.8

4.4

6.6

14.3

8.9

6.6

91.6

96.6

98.2

7.4

4.6

3.4

13.6

6.4

7.4

92.2

109.8

107.5

8.0

3.2

3.0

10.8

5.6

5.4

108.24

105.72

116.41

12.3

7.9

9.7

13.9

8.5

11.2

118.5

110.2

117.0

139

Table 5. Accuracy, intra- and inter-day precisions of the pesticides in fruit and vegetable samples (cont’d)

Pesticides Added

(µg/kg)

Apple Tomato Cucumber Cabbage

Intra

(%)

Inter

(%)

Accuracy

(%)

Intra

(%)

Inter

(%)

Accuracy

(%)

Intra

(%)

Inter

(%)

Accuracy

(%)

Intra

(%)

Inter

(%)

Accuracy

(%)

Thiobencarb 50

100

150

8.2

5.6

4.9

8.2

6.5

5.6

102.4

104.3

106.0

9.50

6.62

3.49

13.0

7.0

6.0

89.1

91.6

101.6

9.8

8.5

5.9

11.9

10.5

6.6

98.9

109.6

113.1

11.8

9.1

5.9

12.1

9.8

6.6

108.9

113.3

113.5

Quinalphos 20

50

100

8.9

7.5

3.9

13.2

11.2

4.4

105.8

89.0

100.6

6.31

9.16

3.36

12.1

11.0

8.2

86.5

88.5

103.3

11.7

10.4

8.3

12.7

11.3

8.9

102.7

115.1

112.7

11.0

11.1

4.5

12.4

13.0

7.5

88.3

108.6

111.5

Endosulfan I 50

100

150

5.2

6.0

4.6

15.5

6.8

5.1

99.1

102.4

109.3

11.83

7.03

1.93

12.0

8.0

6.7

87.5

98.2

98.3

8.3

6.4

6.0

9.3

7.2

7.1

80.8

89.5

87.9

10.0

7.2

4.4

10.2

9.2

4.9

96.8

94.6

90.0

Endosulfan II 50

100

150

7.8

5.8

3.3

8.2

6.5

4.2

90.9

99.8

102.4

5.30

2.68

2.74

5.8

3.9

3.0

87.0

96.2

95.4

10.3

7.5

6.2

10.8

8.3

6.4

76.7

81.8

85.6

11.0

9.7

5.3

11.9

10.8

6.2

96.7

97.1

107.5

Bifenthrin 50

100

150

6.4

6.5

4.1

7.4

6.5

4.1

101.1

106.2

104.9

6.84

4.3

1.56

9.3

6.0

5.3

90.9

96.6

90.9

12.4

6.9

6.4

13.1

7.8

6.7

84.7

83.4

87.2

10.3

6.4

6.8

13.3

7.0

7.2

91.3

88.8

98.5

Fenpropathrin 5

10

20

10.8

10.0

10.7

10.6

10.8

11.4

94.3

102.8

98.2

7.21

4.95

7.09

9.1

9.1

8.3

94.8

97.3

97.8

13.9

11.7

7.0

14.2

12.5

8.1

77.4

83.3

87.2

12.5

11.2

8.6

14.4

14.1

9.6

106.9

90.7

113.2

Permethrin 20

50

100

8.7

5.8

3.4

8.8

6.4

5.01

101.6

98.8

103.5

11.74

6.18

4.71

13.1

7.2

14.7

102.3

104.9

98.4

11.6

7.3

5.1

12.8

10.9

7.7

111.3

108.5

110.6

11.0

10.5

3.4

12.4

12.0

4.6

112.8

88.3

112.2

140

Table 6. Figures of Merit of the Developed Method

Pesticides Apple

(µg/kg)

Tomato

(µg/kg)

Grape

(µg/kg)

Cucumber

(µg/kg)

Fenobucarb LOD

LOQ

MRL

2.41

8.03

300

2.49

8.33

1000

2.17

7.22

300

1.74

5.81

300

Ethoprophos LOD

LOQ

MRL

1.31

4.36

20

0.23

0.77

20

1.20

4.00

20

0.35

1.15

20

Diazinon LOD

LOQ

MRL

0.88

2.92

10

0.21

0.68

10

1.05

3.50

10

0.32

1.05

10

Chlorothalonil LOD

LOQ

MRL

2.16

7.21

1000

6.94

23.12

2000

0.43

1.44

10

8.33

27.76

1000

P. Methyl LOD

LOQ

MRL

0.24

0.79

10

0.62

2.24

10

0.22

0.72

10

0.50

1.65

10

Fenitrothion LOD

LOQ

MRL

0.53

1.77

10

1.35

4.48

10

0.20

0.66

10

0.26

0.85

10

Chlorpyrifos LOD

LOQ

MRL

3.30

11.01

500

3.71

12.36

500

2.79

9.29

500

2.96

9.87

500

Thiobencarb LOD

LOQ

MRL

3.48

11.58

100

4.34

14.47

100

3.19

10.62

100

4.03

13.43

100

Quinalphos LOD

LOQ

MRL

2.16

7.37

50

1.94

6.48

50

2.24

7.47

50

1.94

6.48

50

Endosulfan I LOD

LOQ

MRL

2.30

7.67

50

3.91

13.14

50

3.45

11.50

50

3.25

10.83

50

Endosulfan II LOD

LOQ

MRL

2.17

7.23

50

3.19

10.63

50

3.28

10.95

50

2.08

6.92

50

Bifenthrin LOD

LOQ

MRL

0.11

0.38

300

0.99

3.31

300

0.75

2.50

100

0.89

2.96

300

Fenpropathrin LOD

LOQ

MRL

0.14

0.47

10

0.52

1.72

10

0.55

1.83

10

0.75

2.50

10

Permethrin LOD

LOQ

MRL

1.01

3.36

50

1.50

5.00

50

1.94

6.44

50

2.42

8.05

50

141

Des. time

Salt add

Des. temp

Stirring

pH

Ext. time

Ext. temp

181614121086420

Te

rm

Standardized Effect

2.12(response is TCPA, Alpha = 0.05)

151050-5

99

95

90

80

70

60

50

40

30

20

10

5

1

Standardized Effect

Pe

rce

nt

Not Significant

Significant

Effect Type

pH

Stirring

Ext. time

Ext. temp

(response is TCPA, Alpha = 0.05)

a b

142

Fig 1. Plackett-Burman design plot (a) Pareto chart of standardized main effect (b) Normal plot of standardized main

effect (c) Main effect plot (d) Residual plot

6030

200

150

100

6030 105

600300

200

150

100

84 105

270250

200

150

100

Ext. temp

Me

an

Ext. time Salt add

Stirring pH Des. time

Des. temp

Fitted Means

50250-25-50

99

90

50

10

1

Residual

Pe

rce

nt

25020015010050

30

15

0

-15

-30

Fitted Value

Re

sid

ua

l

24120-12-24

4

3

2

1

0

Residual

Fre

qu

en

cy

24222018161412108642

30

15

0

-15

-30

Observation Order

Re

sid

ua

l

Normal Probability Plot Versus Fits

Histogram Versus Order

c

a

d

143

Fig 2. Desirability response surface plot from CCD design (Ext. temp vs. Ext. time)

144

Fig 3. Response optimizer for optimized parameters

CurHigh

Low1.0000D

New

d = 1.0000

Maximum

TCPA

y = 163.0587

1.0000

Desirability

Composite

2.0

10.0

150.0

750.0

15.0

75.0

15.0

75.0Ext. tim Stirring pHExt. tem

[62.4863] [34.0710] [351.6393] [6.0729]

145

Fig 4. GC-MS Chromatogram of Aqueous Sample spiked at 50 µg/kg; 1. I.S (Internal Standard; 2.

Fenobucarb;3. Ethoprophos; 4. Diaxinon; 5. Chlorothalonil; 6. Parathion Methyl; 7. Fenitrothion; 8.

Chlropyrifos; 9. Thiobencarb; 10. Quinalphos; 11. Endosulfan I; 12. Endosulfan II; 13. Bifenthrin; 14.

Fenpropathrin; 15. Permethrin

TOC Graphic

Reprinted (adapted) with permission from (45

). Copyright (1994) American Chemical Society

146

Session IV: Key success for Good Agricultural Practices (GAP)

147

THAI GOOD AGRICULTURAL PRACTICE

Roongnapa Korpraditskul1 and Chainarong Ratanakreetakul

2

1 Research and Development for Agricultural Commodity Standard Center, Faculty

of Agriculture, Kasetsart University, Kamphaengsaen, Nakhonpathom, Thailand 2Department of Plant Pathology Division, Faculty

of Agriculture, Kasetsart University, Kamphaengsaen, Nakhonpathom, Thailand


ABSTRACT

In Thailand, GAP; Good Agricultural Practice for fresh fruit and vegetable (FFV) had been

actively strengthened during 2004-2012. During that period, GAP provided by Ministry of

Agriculture and Cooperatives was developed to support government policy: kitchen of the

world. Global organization such as FAO/WHO approached DOA to implement knowledge

structure of risk assessment, HACCP, GMP and GAP into the production flow from farm to

table. At the same time GLOBALGAP (EUREPGAP), a private standard for retailer and

supplier in European market had been established to prevent unsafe fresh products from farm

to shelf. Thai exporters of FFV to EU market linked to supply chain and must comply

withthese farm gate standards. Components of the standard ensure farm production concerns

food safety, environmental conservation, welfare of working condition and traceability. In

2007, cluster initiatives of western GAP were developed in compliant to GLOBALGAP

standard under the collaboration of various organizations of which administrative offices are

located in Kasetsart University, Kamphaengsaen Campus. Western GAP guideline has

renamed to THAIGAP. Principles and content of private and public standard are compared

in this study. The challenging strategy is to keep smallholder farmers attached to a supply

chain. Thus, THAIGAP standard was used as a key tool for SME (Small Medium

Entrepreneur) to access global market. Thai Q GAP (Q is for quality), government standard,

belongs to Ministry of Agriculture and Cooperatives, had been actively implemented during

2004-2012 as well. Current number of Thai Q GAP certificate issued is reported.

Nevertheless, being sustained in high end market, linking in value chain is one key element as

well as supporting system i.e., service of inputs, farm advisor and third party certifier for

GLOBALGAP/THAIGAP. Development process for extending the best practice of

smallholder farmer was analyzed for key success factors and the gap along the value chain. It

was found that smallholder farmer groups gained knowledge of good practice are willing to

get certification under conditions of market pricing and sustainability. Up to now, THAIGAP

standard is strongly involved and linked to domestic market. Thai Q GAP standard is

mandatory for FFV export to some countries. The current status of THAIGAP standard for

domestic hypermarket and implementation will be stated. This studyaimed to identify the key

success factor of using private standard to build capacity of smallholder farmer group.

Pyramid of GAP development, strategy of public- private standard for sustainability of Thai

GAP are also reported in this paper.

Keyword: THAIGAP Standard, Small Holder Farmer, Building Capacity, Pyramid for

Development

148

INTRODUCTION

Thailand’s agricultural sector plays an important role in the country’s economy in terms of its

GDP distribution and export earnings. In 2003, Thai government announced the national food

safety policy under two authorizations i.e., Ministry of Agriculture and Cooperatives and the

Ministry of Public Health. In 2005, the “Framework on Monitoring and Control of the

Quality of Agricultural Commodity and Food” was formulated. Meanwhile, the concept of

food safety from farm to table has been included in the plan of both ministries. Ministry of

Public health controls food for domestic consumption while Ministry of Agriculture takes

responsibility for trade and export of agricultural products. Thai agricultural commodity

standards were announced continually to facilitate trade. Good Agricultural Practice (GAP)

was announced as a standard for producers and be promoted ever since .In 2010, Thailand’s

exported agricultural products were valued at THB 1,099,035 million (USD 28,044 million),

accounting for 18% of the total exports and making Thailand one of the biggest agricultural

exporters in the world. Among the agricultural products, fresh fruits and vegetable (FFV)

play an important role as basic raw materials in agribusiness sectors for primary processed

products or fruit in can.There are approximately 1 million small farms farming in FFV sector

for the Thailand agribusiness. To keep FFV sectors sustainable in the world market, one of

the more viable strategies is to differentiate FFV as a safe and reliable agricultural product

with high quality. This action enables Thailand to increase and maintain its market share;

especially in EU countries where require higher food safety, environment and working

conditions ‘standards. GLOBALGAP is considered to be an effective and internationally

recognized standard to access the European market. GLOBALGAP is an independent

business to business certification system founded and managed by the European retail

industry as a way of assuring themselves and their customers’ basic food safety standards.

Globally recognized GAP system certification was proposed as shown in Fig. 1. At farm

level, the basic requirement of food safety must be announced across the country. To access

hypermarket, quality and add-ons are included in the requirements. Then, the international

markets require product certification by independent body complying international standard.

149

Fig. 1. Pyramid for GAP development of smallholder farmers and step up to achieve high

end market

The high cost of getting certification is a major drawback to achieve better market access,

especially for small scale farmers. In 2008, the activities of increasing service capability to

support larger scale farmer groups’ linked the groups to exporters and GLOBALGAP

certification needed. Key players in this value chain are the farmers, exporters, buyers, and

the quality infrastructure service providers like those who train farmers and the certification

bodies. These interventions are to support and increase effective capacity of the service sector

serving the certification needs of the FFV sector. In 2013, THAIGAP standard announced the

domestic concern rather than international market. Currently, THAIGAP standard is

implemented locally to elevate SME groups. This study aimed to identify the key success

factor of using private standard to build capacity of smallholder farmer group.

METHODOLOGY

Public-private GAP standard: rational and development of private standard setting were

collected from the starting point of cluster development (Cluster of Western GAP). The

technical content and relevant stakeholder of THAIGAP standard has been described as well

as the difference between of GLOBALGAP and THAIGAP standards. The content and

comparison were summarized in Table.

Process of developing farmer group to comply private standard requirements: Six

groups of producers linked with exporter were interviewed to understand the cost of

GLOBALGAP certification, and reasons for GLOBALGAP Standard adoption and neglected

adoption. Also they were interview on smallholder farmer group capacity building process,

knowledge management, experience sharing, and lessons learn from the cluster

competitiveness and collaboration.

GG/THAIGAP,

Organic

Thai Q GAP, THAIGAP domestic

Platform Food Safety (FFV)

3rd party Certification

Accreditation

3rd party Certification

Awareness and

achievement

150

RESULTS AND DISCUSSIONS

Public-private standard development: The crisis of mad cow disease or bovine spongiform

encephalopathy (BSE) incidence in England and Ireland reported in 1980s, andpeaking in

1993. These incidences had great impact on European community to launch Food Safety

Law, which strictly required traceability system to prevent and protect consumers from

unsafe food. On December 2003, BSE has been also found in a dairy cow in state of

Washington, USA, which forced the European communities to set their own standard to

ensure that all food are safe, from farm to table. GLOBALGAP is one of the most influential

private standards in the area of food safety, traceability and sustainability. GLOBALGAP is

an independent verification system for GAP as a base for supplier compliance.

GLOBALGAP began as Europa in 1997 as a non-profit organization with an initiative by

several European market chains. One important reason for the effort to set up the

GLOBALGAP is the Food Safety Act imposing more liabilities on retailers in terms of food

safety called due diligence. As a consequence, retailers are responsible for the inputs used for

the branded commodities and unbranded FFV is regarded as a brand of the retailer.

GLOBALGAP was developed by retailers for procuring foods that meet the requirements

based primarily on the European agricultural policy. This EU policy is to promote agriculture

in responsible to sustainable development with giving attention for natural resource

utilization most effectively. The policy was also used to give subsidies to producers who

implement measures to protect the environment. If farmers want to receive subsidy under the

policy, farmers need to fulfill a mandatory criteria such as to keep land in good agricultural

condition and care for environment. In Thailand, initiative program of western cluster for

GAP was started in 2004 with the collaborations of FFV exporters, Kasetsart University, and

provincial technical officers. In 2007, Western GAP requirement changed its name to

THAIGAP. THAIGAP activities were performed by 3 partners: Thai Chamber of Commerce,

National Food Institute, and Kasetsart University. During 2009-2010, the benchmarking

process of THAIGAP with GLOBALGAP was carried out emphasizing on approved

modified checklist and the process has been completed in 2010.

Food safety control system in Thailand involved with different departments and different

ministries. Government sectors under Ministry of Agriculture and Cooperatives promote

GAP certificate which called Thai QGAP. Thai Q GAP for food crops was categorized in the

scope of herbal/medicinal plants, field crops, cutting flowers, fruit, and horticulture crops.

Thai Agricultural Standard (TAS) of food crops or GAP food crops newly issued and

announced for completely alignment with ASEAN GAP, the content consist of a pillar of

food safety, quality, welfare and environmental concerns (TAS, 2013).

GAP mentioned in TAS 2013 was grouped in 8 sections: 1. Water source, 2. Site History,

3. Pesticide usage, 4. Quality Management, 5. Harvesting and Produce Handling, 6.

Storage, transporting in farm, 7. Personnel hygiene, and 8. Record keeping and traceability.

THAIGAP, a private standard for domestics, maintained a similar structure of

GLOBALGAP/THAIGAP checklist. There are 3 modules: 1) All Farm Base Module (AB),

the foundation of all sub-scopes defining all the requirements that all producers must first

comply with to gain certification; 2) Crop Base (CB) scope, the clear criteria based on the

food production; 3) FFV Sub-scope Module, the control points and compliance criteria

(CPCC) covering all the requirements of FFV in the supply chain. The number of

GLOBALGAP certified farms in the scope of FFV decreased from 900 farms in 2009 to 300

151

farms in 2012 (Annual report by GLOBALGAP). Re-benchmarking of THAIGAP to

GLOBALGAP was suspended according to the constraint of decreasing numbers of

GLOBALGAP certified farms, especially smallholder farmer as a group certification. It is

also noted that, although the number of GLOBALGAP certified farms has deceased, the

export value of FFV has not decreased.

After completion of benchmarking for export to global market, THAIGAP standard was

rearranged to be more practical for domestic market. In 2014, THAIGAP standard for

domestic was prepared in collaboration with a group of suppliers and Certification bodies. In

order to promote local producer to use the standard as a tool to access retailer and

hypermarket, Thai Chamber of Commerce (standard owner), plays an active role to expand

the implementation and adoption of THAIGAP standard for domestic market.

According to Thai Q GAP, the numbers of certified farms in the whole country are more

than 140,000. Report of certified farm in 2015 (DOA, 2015) showed that number of certified

farms produced longan, mangosteen and oil palm are 35,125, 8,259 and 6,387 farms,

respectively. The numbers of Thai Q GAP certified farms are required for exporters. Recently,

the movement of food safety for FFV in Thailand is active under the collaboration of public

and private stakeholders. THAIGAP domestic standard for retailers in Thailand has launched

for implementation. Initiative project of domestic THAIGAP has adopted in 18 suppliers. All

producers /suppliers supply products to Retailers such as Makro, Tesco Lotus, Tops

supermarket etc. It is noted that the verification system is carried by certification body and

certificate will be issued by THAIGAP Institute. Requirements of THAIGAP standard for

domestic and international markets were shown in Table 1.

Table 1. Comparison of THAIGAP for domestic market and THAIGAP for international

market

THAIGAP domestic THAIGAP/GLOBALGAP

General Regulation ruled by Thai Chamber

of Commerce General Regulation ruled by Food Plus

All Farm Base 27 51

Crop Base 83 113

Fruit and Vegetable 57 70

Traceability (QR code) Traceability (QR code)

Certification Body of ISO 17065 Certification of ISO 17065 GLOBALGAP

approval

Process of developing farmer group to comply private standard requirements:

Development process of smallholder farmer group was learned from smallholder

farmer group implementing GLOBALGAP. At the beginning, linkage between farmer and

exporter must be clarified for responsibility to participate and support the activities through

the project. In the first phase, 3 farmer groups were able to be certified with GLOBALGAP

152

option 2. The scaling up phase was continued to outreach all activities to increase FFV sector

to access higher value markets in EU. It is clearly noted that service providers such as farm

advisors, internal auditors and farm inspectors were required (Fig. 2). For farmer group,

investment of infra-structure, and training and certification cost were major burden to

smallholder farmer. In most cases, exporters paid for certification cost and provided farm

advisors to implement and train farmers. GLOBALGAP option 2 is a tool for quality

management across the whole groups. In general, farmers or producers of vegetables and

fruits owned small land (less than 1 ha.), especially baby corn, chilli pepper, and etc.

Suppliers can play many roles such as being a farmer (run their own farm) as well as

collecting various types of products from various locations of different farms. It is also found

that collectors and suppliers along the supply chain should play participatory role as the

quality assurance to farm practice, when they buy produce directly from farm. For

smallholder farmers, there should be someone or coordinator to manage production plan and

provide understanding of practice on farm (Korpraditskul, et al., 2010).

Fig. 2. Farmer group must be linked with exporters for GLOBALGAP standard certification

Investment cost for GLOBALGAP adoption for 64 farmer members was shown in Table

2. The crop types are Pomelo, Durian, Roseapple, Leafy vegetable, and Mangoesteen. To

comply with GLOBALGAP standard, farm must keep pesticides in a safe and secure place.

Those incur expense, such as toilets and hand washing facilities, living quarters for workers,

pesticide disposal pit, resting area, cloth and storage for protective clothing etc. There are big

variations of investment cost according to the baseline of conceptual consideration of each

member (data not shown here). For example, one producer of Durian farm was willing to

build the pesticide storage and fertilizer storage whereas another one may just modify the

existing building.

Supplier/export

er

Farm Advisor

Internal auditor

Training

Infrastructure

Supporting fund

QMS

153

Wattanawekin (2011) presented reason of adoption of GLOBALGAP standard in target

farmer group. The motivation of farmers to adopt GLOBALGAP showed that they pay most

attention to the quality of the produce. Fifty percent of farmers agreed that GLOBALGAP

standard enhances family and farm workers’ health. However, farmers do not agree that the

produce under GLOBALGAP certificate will be able to bargain price or buyer will offer a

price premium (Tables 2 and 3).Several reasons of non-adoption of GLOBALGAP standard

by the farmers were also interviewed. It was found that once they already get experience in

practice to comply standard, the top rank of the reasons for non-adoption is the absence or

discontinuation of support. However, only one repliesthat the reason is the difficulties with

record keeping.

Ratanakreetakul et al. (2008) showed the difference of sale price between GLOBALGAP

certified and Thai QGAP certified asparagus farms; GLOBALGAP certified asparagus has

13.2% higher sale price than Thai Q–GAP certified one. Price incentive was a significant

factor to set effective quality management system (QMS) within the farmer group. This

investment cost for implementing quality management system includes activities on a)

traceability and understanding, b) Manage all training cost, c) Sample analysis, d)

Administration, and e) Organizationof third party audit. The payback period for these

investmentsis 1.07 year. This report also mentioned that GLOBALGAP certified

farmerswereable to access broader market levels and worldwide than Thai QGAP certified

farmer.

154

Table 2. Investment costs for GLOBALGAP adoption

Investment items N Mean

(USD)

Standard

deviation Min

Pesticide storage/ Fertilizer storage 56 22,577 62307 0

Toilets and hand washing facilities 38 6,610 10074 300

Living quarter for workers 27 14,022 38066 45

Pesticide disposal pit, grading shed, resting

area 22 10,524 25476 150

Protective clothing and storage for

protective clothing 22 3,125 4314 250

Signs, registration fee, trainings, other 4 5750 5560 0

Wattanavaekin, 2011

Table 3. Farmers’ motivations to adopt GLOBALGAP

Motivations N=59 %

Increase the quality of the produce 55 93.22

Enhance family’s and farm workers’ health 50 84.75

Make finding buyers easier 49 83.05

Enhance management practices 49 83.05

Decrease costs for chemicals 48 81.36

Increase access to high-value markets 44 74.58

Buyer offered a purchase guarantee 40 67.80

Buyer required GlobalGAP 32 54.24

Enhance reputation 31 52.54

Buyer offered a price premium 28 47.46

Enhance bargaining power 28 47.46

Wattanavaekin, 2011

155

Comparison study of GAP standard proposed 5 key issues as a platform for producers,

and one of 5 key issues is management system, which consists of record keeping, internal

assessment and corrective action, complaint and recall procedures (Korpraditskul et al., 2010).

It is necessary to implement quality management system in farmer group. There is still a need

to study the relationship and roles of collectors and suppliers in the market chain. It is

suggested that best practice for farmer and collector should be simply clarified with

understanding. Farmers or producers are willing to follow good practice for the sustained

price guaranteed and market. When buyer demands for GAP certificate for export, farmers

are also obligedto have Thai Q GAP (in a significant numbers), this certificate is issued by

Ministry of Agriculture and Cooperatives. Compared with GAP case in Japan, Ministry of

Agriculture, Fishery and Forestry (MAFF) promoted adoption of GAP in Japan in 2007.

MAFF considers GAP as the best practice, which means that farmers and producers are

allowed to apply on their own justification and no need to be certified by the nation (but must

comply with Food Safety Law). The verification of GAP is based on self-assessment.

Adoption of GAP in Japan, JA (Japanese agriculture cooperation) has promoted their own

GAPs. JA organizations have group members. About 50% of agricultural products produces

in Japan are through local JA. It is said that members of JA are all contract farmers. Local

divisions of JA are responsible for collecting produce from farm and distributing them to

fresh market or processed factory (as a supplier or subcontractor) (Nabeshima et al., 2015).

Therefore, it is noted that market categories are involved directly with promoting quality of

produce and its safety in both domestic and international market. Again, pyramid for

development of GAP will offer the role of provincial responsibility to implement

understanding of food safety and work for risk assessment of food safety situation on their

area. Sufficient but simple knowledge of best practice for food safety at farm production must

be available. Beyond farm or ex-farm gate, buyers or collectors always have significant

influence on farmers’ practice. Generally, Thai exporters provide technical advisors who are

responsible for quality management and required document preparation for traceability and

food safety i.e., control of pesticide use, withholding period, and maximum residue limits.

THAIGAP private standard owned by Thai Chamber of Commerce showed clear strategy to

promote this domestic standard together with traceability system (QR code) along the retailer

supply chain. Technical advisors are available to provide general documents and work

instruction or training materials. For a group of smallholder farmers, quality management and

control within the group must be installed as well.

CONCLUSIONS AND SUGGESTIONS

There are 2 GAP standards in Thailand; one belongs to Ministry of Agriculture and

Cooperatives (Thai Q GAP), and the other belongs to Thai Chamber of Commerce

(THAIGAP). Platforms of the two standards are mainly similar under food safety, quality,

worker health and welfare, and environment. However, traceability is strongly concerned for

THAIGAP private standard with QR code. The development of group certification and

quality management system must include service providers, technical knowledge and

training. The most significant factor for 2 GAP standards’ adoption is actually the

sustainability of market or buyers (who require standard). Certification cost is a burden for

smallholder farmer, therefore, it is recommended to evaluate risk and seek professional

assistance for quality and food safety control.

156

REFERENCES

Department of Agriculture, 2015.accessed to http://gap.doa.go.th/ (In Thai)

Korpraditskul, R; Suwannamook, S, Adulyarattanapan, S and Damsiri, W, 2010.Comparison

study of GLOBALGAP, IGAP and ASEANGAP standard, Research and Development

for Agricultural Commodity Standard Center. 173 pp

Korpraditskul, R., Ratanakreetakul, C, Aroonrungsikul and Taenkam, P., 2010. Scaling up

the Farmer Groups producing Fruit and Vegetable under GLOBALGAP Standard option

2, Cluster of Western GAP, 119 pp.

Nabeshima, Kaoru, Michida, Etsuyo, Houng Nam, Vu and Suzuki, Aya, 2015; Emergence of

Asian GAPs and its relationship to GLOBALGAP ; Institute of Developing Economics,

IDE DISCUSSION PAPER No. 507

National Bureau of Agricultural Commodity and Food Standards (ACFS), 2013, Thai

Agricultural Standard (TAS 9001-2013) : Good Agricultural Practices for Food Crop

Rattanakreetakul, C; Aroonrungsikul, C and Korpraditskul, R; Asparagus farm practical

against a high GAP standard: Practice and Transformation cost . J. of ISSAAS vol. 15 No.

1: 249-250 (2008)

Wattanavaekin, W; presentation at THAIGAP seminar 25 May, 2011at Jupiter 12, Impact

Arena, MuangtongThani, Bangkok, Thailand

http://gap.doa.go.th/

157

DEVELOPMENT OF GOOD AGRICULTURAL PRACTICES (GAPs)

MODELS FOR TEA, RICE AND VEGETABLES IN VIETNAM

Dao Bach Khoa*, Nguyen Thi Nhung, Nguyen Van Liem,

Nguyen Ba Huy, Hoang Thi Ngan.

Plant Protection Research Institute (PPRI), Vietnam Academy of Agriculture

Science (VAAS)


ABSTRACT

Elimination of pesticides and heavy metals residues in agricultural products is key to

successful adoption of Vietnamese Good Agricultural Practices (VietGAPs). However, abuse

of pesticides and fertilizers in agricultural production still exists in some crops. The objective

of this study is to explore the application of VietGAPs in reducing the using of pesticides and

fertilizers during the production of rice, vegetables and tea in Vietnam. The results of survey

and analysis showed that soil and water in the rice fields of Bac Ninh province, vegetable

farm of Hanoi and tea farm in Phu Tho provinces were not contamination of pesticides and

heavy metals. By training and guidance of farmers for VietGAPs implementation, the number

of pesticides and chemical spray is reduced 2-3 times per crop season. This also reduced the

amount of chemical fertilizers and increasing organic fertilizers in the models. Finally, the

GAPs manual should be simplified to suit each GAPs crop for farmers to easily understand

and apply.

Keywords: Good Agricultural Practices (GAPs), pesticides and fertilizers, rice, vegetables,

tea, Vietnam

158

INTRODUCTION

Over the past decade, food production has markedly changed on area, yield, production, types

and especially in the requirements for food safety in Vietnam. Food safety has been a major

concern for Vietnamese central and local government and has led to the decree No. 379/QD-

BNN-KHCN that established Vietnamese Good Agricultural Practices (VietGAPs) on 28

January 2008 by the Ministry of Agriculture and Rural Development (MARD). The aim of

VietGAPs is to prevent and minimise the risk of hazards which may occur during production,

harvesting, and post-harvest handling of crops. VietGAP was developed based on

GlobalGAPs and provide standard for: a) site assessment and selection, b) planting material,

c) soil and substance management, d) fertilizers and soil additives, e) water and irrigation, f)

crop protection and use of chemicals, g) harvesting and post harvest handling, h) waste

management and treatment, i) workers’ health and welfare, and j) record keeping, traceability

and recall (VietGAPs, 2008).

MARD has issued the farming products which include vegetables, fruits, tea, coffee,

pepper and rice. These agricultural products have completed the certification process and

enjoyed a number of policies for supporting good agricultural practices application.

According to statistics of MARD, total rice production area in Vietnam is about 7.89 million

ha, of which about 550 ha have been certified by VietGAPs. Total vegetable production is

estimated to be about 735,000 ha, of which around 62,503 ha have been certified by

VietGAPs. Total tea production is estimated to be about 140,000 ha, of which about 400 ha

have been certified by VietGAPs (MARD, 2013).

There were many documents that were issued regarding policy and management of food

safety, but actual food poisoning still occurs which is caused by residue of chemicals and

harmful microorganisms in food. So, the propaganda and guidance of farmers who practice

good agriculture to produce safe products is still being implemented in Vietnam. Three

models of VietGAPs were conducted in rice, vegetables and tea by the Plant Protection

Research Institute (PPRI) in collaboration with the local agricultural extensionists and

farmers in the Northern part of Vietnam.

METHODOLOGY

Three local locations in Nothern of Vietnam were selected for the development of VietGAP

models, including: 50 ha rice (Oryza sativa) in Yen Phu village - Yen Phong district - Bac

Ninh Province; 15 ha vegetable at Linh Nam village - Hoang Mai ward - Hanoi City; and 25

ha tea (Camellia sinensis (L) O. Kuntze) at Phu Ho village - Phu Tho town - Phu Tho

province. Fertilizers (chemical fertilizers and organic fertilizers), pesticides (chemical

pesticides and biological control agent).

The pre-survey was conducted by interviewing 30 farmers, local agricultural extensionists

in each local area regarding information on rice, vegetables and tea production, especially

focusing on technical application including fertilizers and pesticides. Compilation of

technical documentation for farmers and local extentionists about VietGAPs implementation

was also done. Testing of heavy metals and pesticide residues in soil and water before

159

conducting models and post-harvesting of agricultural products were conducted by using

GC/MS, HPLC and AAS.

RESULTS

Survey of current status of areas for VietGAP implementation in rice, vegetables and

tea production.

Location for rice production following VietGAPs standards, about 50 ha, belong to Yen Phu

village - Yên Phong district - Bac Ninh province in Red river delta, total area for rice

production about 293 ha, with two seasons per year. Water, irrigation and local transportation,

pesticides and fertilizer storages are suitable for requirements of VietGAP sstandard. The

heavy metals and pesticide residue analysis of soil and water in the area for VietGAPs

implementation showed under standard level of National technical regulation on the

allowable limits of heavy metals in soils (QCVN 03:2008/BTNMT) and the National

technical regulation on the allowable limits of heavy metals in water (QCVN

039:2011/BTNMT) (table 1,2).

Table 1. Results for analysis of heavy metal in water samples in Yen Phu - Yen Phong - Bac Ninh

No Samples As Cd Pb Hg

1 MN1 0,0005 0,0003 0,0003 0,00003

2 MN2 0,0004 0,0004 0,0005 0,00003

3 MN3 0,0004 0,0004 0,0003 0,00003

Unit mg/kg (mg heavy metal per litter of waterl)

Table 2. Results for analysis of heavy metal in soil samples in Yen Phu - Yen Phong - Bac Ninh


1 MN1 1,68 0,35 27,43 25,30

2 MN2 2,22 0,43 22,18 34,78

3 MN3 2,11 0,40 30,53 17,20

Unit mg/kg (mg heavy metal per kilogram dried soil)

160

Linh Nam village - Hoang Mai ward - Hanoi is a commune with intensive vegetable

production with 100 ha for growing vegetables. Cooperative management board has been

performing a process to produce safe vegetables with high quality products. Farmers have

their own land therefore vegetable safety products depends on the level of technical

application by households. So, the Cooperative has decided to allot 10 ha of vegetables

following VietGAPs model. Infrastructure includes net house to cover rain and sun to

produce leafy vegetables, plastic pine system for irrigation, processing and packaging system.

Farmers have many experiences in production of safe vegetables but they lack knowledge in

VietGAPs, especially the use of chemical pesticides with high rate spay and mixed several

pesticides (Table 3). The heavy metals and pesticide residue analysis of soil and water in the

area for VietGAPs implementation showed under standard level of National technical

regulation on the allowable limits of heavy metals in soils (QCVN 03:2008/BTNMT) and

National technical regulation on the allowable limits of heavy metals in water (QCVN

039:2011/BTNMT) (Tables 4,5).

161

Table 3. List of chemical pesticides use for control pests in vegetable field in Linh Nam – Hoang Mai - Hanoi

Active ingredient

Pests

Striped

flea

beetle

Diamond

back

moth

Oriental

leafworm

moth

Cabbage

aphid

Late

blight

Downy

mildew

Abamectin x x

Azadirachtin x x

Bacillus thuringiensis x

Diafenthiuron x

Dinotefuran x

Emamectin benzoate x x

Etofenprox x

Fenvalerate x

Imidacloprid x

Matrine x

Nitenpyram x x

Permethrin x

Thiamethoxam

Chlorothalonil

Difenoconazole x

Hexaconazole x

Mancozeb x

Metalaxyl-M x

Propiconazole x

Thiophanate-Methyl x

Validamycin

162

Table 4. Results for analysis of heavy metal in water samples in Linh Nam - Hoang Mai - Hanoi


1 MN1 0,0006 0,0004 0,0005 0,00004

2 MN2 0,0005 0,0004 0,0005 0,00003

3 MN3 0,0005 0,0004 0,0004 0,00005


Table 5. Results for analysis of heavy metal in soil samples in Linh Nam - Hoang Mai - Hanoi


1 MN1 1,78 0,45 28,43 21,30

2 MN2 2,28 0,23 20,18 37,78

3 MN3 2,61 0,30 33,53 27,20


Phu Ho village, Phu Tho town, Phu Tho province lead the midlands of the mountains in

the Northern part of Vietnam. Natural condition in Phu Tho province is suitable for growing

tea with a total of 15.700 ha of tea plantation in this region. The area for developing

VietGAPs is around 25 ha, with infrastructure which includes water, irrigation and local

transportation, pesticide and fertilizer storages. The heavy metals and pesticide residue

analysis of soil and water in the area for VietGAPs’ implementation showed under standard

level of the National technical regulation on the allowable limits of heavy metals in soils

(QCVN 03:2008/BTNMT) and National technical regulation on the allowable limits of

heavy metals in water (QCVN 039:2011/BTNMT) (Table 6, Table 7).

163

Table 6. Results for analysis of heavy metal in water samples in Phu Ho - Phu Tho


1 MN1 0,0003 0,0003 0,0005 0,00004

2 MN2 0,0003 0,0004 0,0003 0,00004

3 MN3 0,0004 0,0003 0,0004 0,00005


Table 7. Results for analysis of heavy metal in soil samples in Phu Ho - Phu Tho


1 MN1 1,88 0,35 18,43 24,13

2 MN2 1,21 0,23 21,18 27,43

3 MN3 1,91 0,25 13,53 25,30


VietGAPs models

Training farmers for VietGAPs.

The new technical documentations were released by a combination of local production

process for vegetables and VietGAP standards. Fifteen classes of VietGAPs for rice,

vegetables and tea were organized with 300 famers attending. Farmers understand more

about the technical methods for rice, vegetables and tea safety production, such as: how to

use the manure and chemical fertilizers in products without excess NO3- and without harmful

micro-organism, using pesticides according to four true, no arbitrary increasing the dosage

mixing two or three pesticides in one. Ensure that no pesticide residues exceed within the

permitted level of the FAO/WHO or standard level of Vietnam in rice, vegetables and tea,

and encourage farmers to use biological control agents and only use chemical pesticides in

case it is necessary. The farmers families keep a record of their activities for VietGAPs that

is more easy

164

Development of VietGAPs models for rice, vegetables and tea production.

Fifty hectares of rice with sticky rice seeds, attending 200 households, were cultured

following the VietGAPs standards. Process of fertilizer application for VietGAPs model is

shown in Table 8. The survey results showed that the main pest appeared inside and outside

of models with pesticides and time for spray (Table 9). Chemical pesticides were used for

inside and outside models AS showN in table 9. Analysis of pesticide residue after

harvesting of rice samples is shown in Table 10. Economic efficiency of reduced number of

pesticide sprays between inside and outside VietGAP models is shown in Table 11.

Ten hectares of vegetables, with many kinds, attended by 50 households, were cultured

following the VietGAPs standards. The survey results showed that the main pests appeared

inside and outside models with pesticides and time for spray (Table 12). Economic efficiency

of reducing the number of pesticide sprays between VietGAPs and normal models are shown

in Table 13..

Twenty-five hectares of land planted to tea, attended by 50 households, were cultured

following the VietGAPs standards. The survey results showed that the main pests appeared

inside and outside of models with pesticides and time for spray (Table 14). Table 15 shows

the economic efficiency of reducing the number of pesticide sprays between VietGAPs and

normal models.

Table 8. Process for application of fertilizers in VietGAP model for rice production

Time for application Manure N P K

First 10.000 36 85

Second 36 50

Third 18 50

Total 10.000 90 85 100

Unit kg/ha

165

Table 9. Main pests, pesticide and time for spray in season in rice field

Pest Pesticides

Time of sprays

VietGAP

model

Normal

model

Stemborer (Scirpophaga

incertulas Walker)

Thiamethoxam (Virtako 40WG),

chlorantraniliprole (Prevathon

5SC)

1 1

Brown planthopper

(Nilaparvata lugens Stal)

Nitenpyram (Elsin 10EC) ,

Thiamethoxam (Actara 25WG)

1 2

Rice skipper (Parnara guttata

Bremer et Grey)


Deltamethrin (Ebato 160SC)

1 2

Bacterial blight (Xanthomonas

oryzae)

Xanthomix 20WP, Staner 20WP 1 1

Rice blast (Piricularia oryzae

Cavara)

Hibim 31WP, Beam 75WP 1 1

Sheath blight (Rhizoctonia

solani Kuhn)

Anvil 5SC, Tungvil 5SC 0 0

Table 10. Analysis of pesticide residue in rice samples with VietGAP afer harvesting

Sample

Results

Thiamethoxam Chlorantraniliprole Nitenpyram Fenobucarb Fipronil

M1 nd nd nd nd nd

M2 nd nd nd nd nd

M3 nd nd nd nd nd

M4 nd nd nd nd nd

M5 nd nd nd nd nd

nd: none detection

166

Table 11. Economic efficiency of pesticide using between VietGAP and normal models in

rice

production per hectare

Unit VietGAP model Normal model

Time for sprays (time/ha) 5 7

Pestidice price (1000 VND) 2.000 2.800

Labor for sprays (1000 VND) 2.000 2.800

Total (1000 VND) 4.000 5.600

Save amount (1000 VND) 1.600

167

Table 12. Main pests, pesticide and time for spray in season in vegetable fields

Pest Pesticides

Time of sprays

VietGAP

model

Normal

model

Striped flea beetle (Phyllotreta

striolata)


Nitenpyram (Elsin 10EC)

1 2

Oriental leafworm moth

(Spodoptera litura)

Permethrin (Pounce 50EC),

Fenvalerate (Sudin 20EC), Permethrin

(Perkill 10EC).

1 1

Diamondback moth (Plutella

xylostella)

Bacillus thuringiensis (V-Bt);

Emamectin (Proclaim 1.9EC);

Lambda-Cyhalothrin (Match 50EC);

1 2

Aphids Abamectin (Elincol 12ME);

Etofenprox (Trebon 10EC);

Imidacloprid (Admire 50EC) ;

1 2

Thrip (Thrip palmi) Imidacloprid (Confidor 100SL);

Thiamethoxam (Actara 25WG);

Dinotefuran (Oshin 20WP)

1 1

Death of seedlings (Rhizoctonia

sp)

Hexaconazole (Anvil 5SC);

Chlorothalonil (Daconil 75WP);

Metalaxyl (Ridomil Gold 68WP);

1 1

Late Blight (Phytophthora

infestan)

Metalaxyl (Ridomil Gold 68WP);

Fosetyl Aluminium (Aliette 80WP)

1 1

Black spot disease

(Colletotrichum sp)

Difenoconazole (Score 250EC);

Carbendazim (Bavistin 50SC);

Propiconazole + Difenoconazole (Tilt

Super 300EC)

1 1

Powdery mildew (Podosphaera

xanthii)

Hexaconazole (Anvil 5SC);

Propiconazole + Difenoconazole (Tilt

Super 300EC)

1 2

https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&cad=rja&uact=8&ved=0CCIQFjAAahUKEwi4-LGQ6PTIAhUBqqYKHSqLB4Q&url=http%3A%2F%2Fvegetablemdonline.ppath.cornell.edu%2Ffactsheets%2FPotato_LateBlt.htm&usg=AFQjCNHNcPGz3KnsDBOV8iETNGdwEreu2g&sig2=0KkMyXkYrcqps8YoIg4INA

168


vegetables production per hectare

Unit

VietGAP model Normal model

Cabbage Chinese

cabbage Tomato Bean Cabbage

Chinese

cabbage Tomato Bean

Time for sprays

(time/ha) 4,2 4,1 4,7 6,1 6,1 5,1 5,8 7,8

Pestidice price

(1000 VND) 1.470 1.430 1.645 2.135 2.135 1.785 2.030 2.730

Labor for sprays

(1000 VND) 1.680 1.640 1.880 2.440 2.440 2.040 2.320 3.120

Total (1000

VND) 3.150 3.070 3.525 4.575 4.575 3.825 4.350 5.850

Save amount

(1000 VND) 1.425 755 825 1.275 - - - -

169

Table 14. Main pests, pesticides and time for spray in season in tea fields

Pest Pesticides

Time of sprays

VietGAP

model

Normal

model

Green leafhopper (Jacobiasca

formosana)

Abamectin + Matrine (Sudoku

58EC), Buprofezin + Isoprocarb

(Superista 25 EC), Abamectin

(Song Mã 63EC)

1 2

Mosquito bugs (Helopeltis sp) Azadirechtin (Vinaneem 2 SL),

Abamectin (Acimetin 5EC)

1 1

Red spider mite (Oligonychus

coffeae)

Fenpyroximate (Ortus 5SC),

Pyridaben + Abamectin (Aben

168 EC)

1 2

Thrips (Physothrips setiventris) Abamectin (Abagro 4.0 EC),

Aremec 36EC

1 2

Colletotrichum thaee sinensis Eugenol (Genol 0.3DD),

Trichođerma viride (Biobus

1.00WP)

1 1

Exobasidium spp Masse Imibenconazole (Manage 5WP) 1 1

170


tea production per hectare

Unit VietGAP model Normal model

Time for sprays (time/ha) 7 9

Pestidice price (1000 VND) 500 500

Labor for sprays (1000 VND) 250 250

Total (1000 VND) 5.250 6.750

Save amount (1000 VND) 1.500

CONCLUSION

This study investigates and compares the farming practices of selected farms where three

main crops (rice, vegetables, and tea) are grown under VietGAPs model and those that

practice the normal (traditional) model in the Northern part of Vietnam.

Results of analysis of soil and water showed that most of the area for growing crops in

Vietnam are not contaminated by heavy metals and pesticides.

There are 21 active ingridients of pesticides that are used for control of main pests in rice,

vegetables and tea. The number of pesticide sprays in the normal model are usually higher

than the VietGAPs model that means costs increase for production.

Farmers will successful apply VietGAPs models if there are suitable policies for training

and the information is disseminated in media. VietGAPs should be expanded in large areas

and used for other agricultural crops.

171

REFERENCES

VietGAPS (2008) Good agricultural practices for production of safe fresh fruit and

vegetables in VietNam (VietGAP). Hanoi. Ministry of Agricultrue and Rural

Development, pp23.

Vuong, P.T., N.T. Nhung., N.T. Thuy., D.T. Hang., T.Q.Viet., D.T.Luong and N.T.Q.Trang,

2010. Development of safe vegetable production for improving generation income of

household and protecting of rural environment in Vietnam. Journal of Vietnamese

Agricultural Science and Technology 1 (14). 43-49.

Oleg, N., E.V.D. Fliert., H.V.Chien., V.Mai and L.Cuong, 2010. Good Agricultural Practice

(GAP) as a vehicle for transformation to sustainable citrus production in the Mekong

Delta of Vietnam. 9th European IFSA Symposium, 1893-1901.

Gazi, M.N.I., F.M.Arshad., A.Radam., E.F.Alias, 2012. Good agricultural practice (GAP) of

tomatoes in Malaysia: Evidences from Cameron Highland. Africa Journal of Business

Management. 6(27). 7969-7976.

172

PRODUCERS’ PERCEPTIONS OF PUBLIC GOOD AGRICULTURAL

PRACTICES AND THEIR PESTICIDE USE: THE CASE OF MYGAP

FOR DURIAN FARMING IN PAHANG, MALAYSIA

¹ School of Agriculture, Kyushu University, Fukuoka, Japan; Present Address: 6-10-1

Hakozaki, Higashi-ku, Fukuoka 812-8581 Japan

² Department of Environmental Science and Management, University of Malaya, Kuala

Lumpur, Malaysia

³ Department of Southeast Asian Studies, University of Malaya, Kuala Lumpur, Malaysia

email: [email protected]

ABSTRACT

There have been growing interests in the rise of public Good Agricultural Practices (GAP)

standards in Southeast Asia that have been implemented by the governments in the region.

This paper examines the local implementation of Malaysian public GAP standard called

‘MyGAP’ with a focus on producers’ perceptions of their participation as well as their on-

farm practices for safety assurance. For this objective, producers’ perceptions of the benefits

and shortcomings of the scheme and their pesticide use practices are examined by comparing

the cases of MyGAP certified and uncertified durian farms in the state of Pahang, Malaysia.

The research found that the certified farmers see the usefulness of the MyGAP program

mainly in the minimum securement of opportunities for exporting their produce, and that

overall, certified farms are using significantly less pesticides than those of uncertified farms.

Keywords: MyGAP; public GAP standard; food safety; pesticide use; durian farming;

Malaysia


173

INTRODUCTION

In recent years, the implementation of GlobalGAP and some other private food safety

standards has raised the critical question of how to balance out the imperative of safety and

quality assurance in the production and consumption of food, and the participation in global

value chains of small-scale farmers in the Global South (Amekawa 2013a). For small-scale

farmers in developing countries, GlobalGAP requirements for food safety could be too high

to meet (Amekawa 2009). It is mainly due to costly investments in valuable inputs like

switching to approved pesticides and building long-term structures such as farm toilet and

pesticide storage unit (Okello and Swinton 2007). Since the early 2000s, several development

aid organizations have reported the tendency of small-scale farms in developing countries to

be dropped off or excluded from global value chains (e.g., Asfaw 2007; Mungai 2004;

Graffham et al. 2007). Around 2010, a series of studies started to examine the impact of

GlobalGAP certification on the access to export markets and associated economic conditions

of producers in developing countries. Many of them identified the socio-economic attributes

or characteristics of GlobalGAP certified farms (in most cases through comparison with the

case of uncertified farms); and demonstrated the socioeconomic advantage of GlobalGAP

certified farms over uncertified farms (e.g., Asfaw et al. 2010; Colen et al. 2012; Henson et

al. 2011; Holzapfel and Wollni 2014; Kersting and Wollni 2012; Kleemann et al. 2014;

Tallontire et al. 2013).

Apart from the conspicuous trend of private global food safety standardization led by

GlobalGAP, there is a public stream of food safety standardization in Southeast Asia that

seems to garner academic attention (e.g., Amekawa 2009, 2010, 2013a, 2013b; Banzon et al.

2013a, 2013b; Islam et al. 2012; Mankeb et al. 2014; Nicetic et al. 2010; Pongvinyoo and

Yamao 2014; Pongvinyoo et al. 2014; Schreinemachers et al. 2012, Srisopaporna 2015).

Since the early 2000s, several countries of the Association of South-East Asian Nations

(ASEAN) have introduced national public GAP standards with the objective of improving the

safety and quality of agricultural produce. This is largely a response to the rapidly increasing

levels of agricultural pesticide use in the region, as well as the increasing concerns of foreign

and domestic consumers about food safety (Schreinemachers et al. 2012). It is imperative that

the actual working of public GAP standards in Southeast Asia be studied in light of the

original purpose of food safety assurance as well as the broader goals related to the welfare of

producers and environmental protection. This is said with special regard to the timing when

the regional common GAP standard called ‘AseanGAP’ is to take effect in 2015, with which

each national public GAP standard in the ASEAN region is planned to benchmark.

This paper examines the implementation of Malaysian public GAP standard called

‘MyGAP’ with a focus on producers’ perceptions of their participation in the scheme as well

as their practices for safety assurance. The latter is examined by comparing the pesticide use

and handling between MyGAP certified and uncertified durian farms in Raub District and

Bentong District in the Pahang State of Malaysia. Based on the farm-level case study, it

considers what characteristics MyGAP would have in the ASEAN regional context of food

safety standardization. The paper is organized as follows: The second section introduces the

main features and problems of public GAP standards in Southeast Asia. The third section

provides an overview of the MyGAP program. The fourth section discusses research context

and methods. The fifth section presents research findings. The sixth and last section provides

conclusions.

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PUBLIC GAP STANDARDS IN SOUTHEAST ASIA

Table 1 shows the basic information of national public GAP standards in 6 Southeast

Asian countries that are currently under implementation, as shown along with GlobalGAP

and AseanGAP. While sharing the goal of food safety assurance, public GAP standards in

Asean countries encompass varying levels of grower adoption and differing ways of policy

implementation. Thailand has the largest public GAP program in terms of the amount of

certification. As of August 2015, as many as about 119,000 farms (mostly small-scale),

received a Q-GAP certification (Q refers to ‘quality’). The crude scale of certification even

surpasses GlobalGAP which has wielded a strong influence over global food safety

standardization. Compared with the number of certifications of over 220,000 in 2012,

however, it has rapidly declined since 2013 when the country introduced the new code of

practice (the version TAS 9001-2013), which has made it more difficult for applicant farms

to get a Q-GAP certification. The amount of certified farms in the other listed ASEAN

countries is much lesser due to different levels of stringency in compliance for certification.

By way of illustrating the gap in the level of stringency in compliance, MyGAP requires 95-

100% of compliance for 106 control points out of the total of 163 control points (Department

of Agriculture, Malaysia 2005); in comparison, out of the total of 116 control points, Q-GAP

requires 100% of compliance for 23 control points and 60% of compliance for 41 control

points, along with 54 recommended points (Department of Agriculture, Thailand 2013).

Table 1. Adoption of national public GAP standards in selected countries in Southeast Asia

as shown with the cases of GlobalGAP and AseanGAP

Source: created through reference to the GAP protocol and direct contact to the agency in

charge in Southeast.

In accordance with the significantly limited level of required compliance in Q-GAP,

empirical findings suggest that the quality of food safety assurance is compromised. For

instance, Schreinemachers et al. (2012) compared 45 Q-GAP certified and 245 uncertified

farms for a total of 9 vegetable and fruit crops in a watershed of Chiang Mai Province,

Northern Thailand, only to find that there are no significant statistical differences between the

2 farmer groups in terms of the amount of pesticides used, methods of pest control adopted,

and pesticide handling. Amekawa (2013b) found that 34 of the 64 Q-GAP certified pomelo

growers from 2 communities of Chaiyaphum province, Northeast Thailand, showed a lack of

understanding of the concept of GAP. In addition, most of those who noted a reduction of

Country/region Program Year of Inception

Number of Certified

Farms (year) Responsible Agency

Europe GlobalGAP 1999 112,576 (2011) EurepEuro-Retailers Produce Working Group

Malaysia MyGAP 2002 313 (2013) Department of Agriculture

Thailand Q-GAP 2004 ≒220,000 (2012) Ministry of Agriculture and Cooperative

≒119,000 (2015)

Singapore SingaporeGAP-VF 2004 7 (2013) Agri-Food & Veterinary Authority

The Philippines PhilGAP 2005 15 (2013) Department of Agriculture

Viet Nam VietGAP 2008 575 (2013) Ministry of Agriculture and Rural Development

Brunei BruneiGAP 2013 1 (2014) Ministry of Industry and Primary Resources

Asean region AseanGAP 2015 (planned) T.B.D. Asean Secretariat

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their pesticide use around the period of their certification attributed it to the growth stage of

pomelo rather than the positive effect of Q-GAP certification.

In view of the current situation where research on public GAP standards lags far behind

research on private GAP standards, it is necessary to do further empirical studies on public

GAP programs in Southeast Asia; especially with regard to the current practices of food

safety and quality assurance in food production. Malaysian MyGAP provides an interesting

case in point. The GAP standard has produced the second largest amount of public GAP

certification in Southeast Asia even though it is regarded to embrace the relatively strict

criteria of standard certification. The reality of the compliance of certified farms with

MyGAP standard needs to be examined empirically, however, with regard to its

implementation of food safety and quality assurance. As regards producers’ pesticide use

practices, it is hypothesized in this study that unlike the extant research findings related to Q-

GAP, MyGAP certified farms would have significantly better results than those of uncertified

farms due to its much higher standard of compliance for certification than those of Q-GAP.

MYGAP IN MALAYSIA

In 2002 the Malaysian government established a public GAP certification scheme for

fresh fruits and vegetables called SALM (Skim Akreditasi Ladang Malaysia1)

) or Malaysian

Farm Accreditation Scheme) (Islam et al., 2012; Salleh and Osman, 2007; van der Valk and

van der Roest, 2009) – the original GAP scheme for MyGAP, along with other GAP schemes

for fishery and livestock2 )

, SALM was aimed at creating vibrancy within the domestic

commercial fresh fruits and vegetables (FFV) sector by promoting “agricultural practices that

are environment-friendly, sensitive to workers’ welfare and yield quality products that are

safe for consumption” (Robert and Menon, 2007:31). While the Department of Agriculture

(DoA) serves as the secretariat for MyGAP, it works in collaboration with various state

agencies. Major decisions are made by a steering committee called the National Farm

Accreditation Committee (NFAC), which comprises the representatives from various

government and government-related agencies (van der Valk and van der Roest, 2009). The

Department of Standard Malaysia (DSM) and any agency licensed by the DSM are the

agencies that accredit the farms for good agricultural practices (Salleh and Osman, 2007).

In 2005, SALM-certified farms were entitled to use the ‘Malaysia Best’ logo with their

products, which provides them more marketing advantages (Robert and Menon, 2007). In

August 2013 the Ministry of Agriculture and Agro-based Industry launched MyGAP

(Malaysian Good Agricultural Practices) as the rebranding exercise of the 3 existing GAP

schemes established in 2002. Hence, MyGAP emerged as a comprehensive certification

scheme for the agricultural, aquaculture, and livestock sectors (Ministry of Agriculture and

Agro-based Industry Malaysia, 2014).

1 The Malay denotation of the scheme was later changed to “Skim Amalan Ladang Baik Malaysia,” carrying the same acronym SALM but

with the different meaning of ‘Malaysian Farm Certification Scheme for Good Agricultural Practice Scheme’ (Othman 2006). The change

was deemed necessary because it was realized that “the DoA was in no position to accredit farms complying with the conditions set by the

department” (Salleh & Osman 2007: 46).

2 There were 2 other GAP schemes that were established concomitantly: Malaysian Aquaculture Farm Certification Scheme (SPLAM) by

the Department of Fisheries Malaysia and Good Animal Husbandry Scheme (SALT) by the Veterinary Service Department (Othman 2006).

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The participation of farms in the MyGAP scheme is voluntary. Any individual farm

growing crops of relevance to the economy at large and complying with the initial

requirements for land tenure, the location of the farm, and water sanitation can apply for it.

Upon application registration, the farm is required to conform to a series of requirements

meted out for MyGAP certification (Salleh and Osman, 2007). The MyGAP certification for

agriculture consists of 16 categorical items, each of which comprises specific rules or

conditions based on the Malaysian Standard MS 1784: 2005 – Crop Commodities – Good

Agricultural Practice (GAP) (Othman, 2006). The DoA sends to the applicant farm a team of

auditors who are normally local DoA officers on the daily basis yet could be officers from the

DoA Malaysia from Putrajaya once in 2 years, in order to check if the applicant farm

complies with a set of required control points (interview with DoA officers at DoA Raub on

10 December, 2013). Record keeping is one of the most important elements for farm

verification. Every farm activity should be recorded for the sake of traceability for the

produce or the farm worker (Salleh and Osman, 2007).

Once the results of the verification of farm practices are submitted to NFAC, the produce

and the water from the farm are collected and analysed for pesticide residues and heavy

metals. The samples are taken 3 times over the production season. The 3 samples must not

exceed the defined maximum residue levels (MRL). If any of the samples contain residues

above the MRL, then another 3 separate samples will be collected. Confirming that no

residues exceed the set MRLs, the MyGAP Committee approves MyGAP certification for the

applicant farms. A certification will last for 2 years. Before the end of the term, the farm can

reapply for recertification. For these farms, only 1 sample will be required for residue

analyses instead of 3 (Salleh and Osman, 2007). The government bears the cost of inspection

and residue analysis, providing publicity for promotion (van der Valk and van der Roest,

2009).

RESEARCH CONTEXTS AND METHODS

According to the DoA Malaysia, of the total national certifications of 313 in 2013, durian

comprised the second largest number of certifications after rice. Since our focus for this study

was FFV, we chose durian for the target crop for this research. As of July 2013 there were 21

certified durian farms in Pahang State, out of which 19 were interviewed3)

. The certified

farms were contacted for interview with assistance by local DoA officers and using the local

official DoA directory of MyGAP certifications for durian in Raub District and Bentong

District. For the purpose of comparison, 57 uncertified durian farmers were also interviewed

so that the number of interviewed uncertified farms could triple that of interviewed certified

farms. Reflecting the regional pattern of durian production in the state of Pahang, the

majority of farms interviewed were located in Raub District, the most prosperous durian

production district in Malaysia. In addition, 1 certified and 3 uncertified farms were located

in Bentong District (Fig. 1).

3 Two certified farms were excluded from this research as they were the DoA experimental farms operating on the public basis.

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Fig. 1. Map of Pahang State, Peninsular Malaysia

Source: Ramam et al. (2007)

Raub comprises 7 mukim (territorial divisions), namely Batu Talam, Sega, Semantan Ulu,

Dong, Ulu Dong, Gali, and Tras, in the total area of 2,269km². All the areas of the district are

dominated by Chinese population except for 2 villages such as Kampung Jeru and Sungai

Pasu, where the majority of the population including durian farmers is Malay. The main crops

grown in this area include rice, durian, cocoa, oil palm, and natural rubber.

Certified farms were identified in 6 areas including Sungai Klau, Sungai Ruan, Sungai

Chetang, Pekan Cheroe, Tras, and Raub Trade Center. Uncertified farms were identified in

these areas for interview. With no available official residential information of uncertified

farms along with the limited available span of data collection, however, it was not possible to

implement a systematic random sampling. Instead, with the help of local DoA officers, the

data collectors relied on several local producer groups for snowball sampling in addition to

sporadic farm visits. To avoid a sampling bias, only farms with the size of durian orchard

between 1 and 10 ha were called for interview. Another criterion for the selection was to

focus on those uncertified farms who had never applied for MyGAP at the time of the

research. The interviews were conducted by Chinese and Malay speaking research assistants

in January and July 2013.

The survey questionnaire form was organized into 7 sections: 1. Basic farm

characteristics; 2. Economic and financial aspects of the farm; 3. Perceptions of MyGAP

policy and certification; 4. Training and processes for obtaining certification (for certified

farms only); 5. Experiences of audit (for certified farms only); 6. Pest and crop management;

and 7. Pesticide use and handling. The majority of certified farms comprise a farm manager

who is the owner of the farm and 1 or 2 employed workers for on-farm practices, while most

of the uncertified farms were run by a farm manager and 1 or no farm worker. Most of the

workers are migrant Indonesians, followed by migrant Burmese and local Malaysians. The

farm manger was interviewed on 1 ~ 5 sections (the manager of the uncertified farm was

focused on 1 ~ 3 sections only), whereas the worker was interviewed on 1, 6, and 7 sections

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of the questionnaire. In interviewing individual farmers, notes were taken on the structured

question form and all the interviews were tape-recorded. The length of an interview was

around 60 minutes. Part of the obtained data was structured into the database file and used

mainly for quantitative analysis. Data collectors also recorded any conspicuous qualitative

information in the form of a descriptive summary, at times with direct quotes of the

interviewees’ remarks as translated into English.

RESULTS

Farmers’ adoption of MyGAP

The total land size of certified farms is nearly 90% larger than that of uncertified farms. One

certified farm holds a predominantly large farmland of 42.1 ha (out of which 14.5 ha is used

for durian farming), while the land size of the rest of certified farms is 14 ha or less.

Excluding the farm with 42.1 ha of land, the average size of certified farms shrinks to 5.4 ha.

The total durian farm size of certified farms is nearly 60% larger than that of uncertified

farms, with the number of employed workers of certified farms being 40% more than the

latter (Table 2). This gap in the durian orchard size may be related to the general incentive of

export farming for the adoption of MyGAP since agricultural export generally requires more

economic scale than domestically-oriented transactions (Johnston and Mellor, 1966). The

average number of employed workers on certified farms appears nearly 3 times larger (3.3

versus 1.2, for certified and uncertified farms, respectively). However, if a certified farm with

30 employed workers (who work on the planting of not only durians but also several other

crops) is excluded, the number declines to 1.7. Seventy-nine percent of certified farm

managers have secondary education or higher while 57% of uncertified farm managers had

primary education or less. Such an observed superiority in the educational background of

certified farms over uncertified farms seems to conform to the observations in some literature

of good agricultural practices (e.g., Asfaw et al., 2010; Kersting and Wollni, 2013;

Pongvinyoo et al., 2014).

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Table 2. Background of respondent farms

On the issue of why individual farms have decided to participate in the MyGAP program,

the majority of certified farm managers (77%) responded that they had decided to apply for

MyGAP certification because they expected that once they obtain the certification, they will

be eligible to export their produce to overseas markets. This result comes as no surprise

because in many cases DoA extension officers tell farmers that MyGAP certification is a

minimum requirement for the export of their durian produce. For most durian farm managers,

the farm gate price of durian is their utmost concern since durian sales are their main (or only)

source of income. Their main goal is to be able to export durians in order to improve their

economic conditions. They claimed that it is difficult to negotiate on the prices, however, as

middlemen are positioned to play much more powerful roles in controlling farm gate prices

by maximizing their profits through selling produce to the market at much higher prices.

Apart from economic motives, only 5 farm managers (26%) considered the improvements in

food quality assurance as a reason for their participation in the MyGAP scheme. In this

regard, a significant gap in the expectations of farmers and local DoA officers was observed.

Farmers claimed that economic expectations for MyGAP must be their primary concern of

their participation in the food safety program as they make a living based on farming.

However, DoA officers seemed to undervalue this point. They complained that farmers do

not understand the main objective of the policy which is food quality assurance through

conducting good agricultural practices, and that they rather insist solely on economic interests

instead.

The primary reason why uncertified farm managers had not applied for MyGAP at the

time of research is that they had little or no knowledge about it. Thirty-seven uncertified farm

managers (74%) responded that they had never heard about it and thus had no clues for

Certified Uncertified

Total number of studied farms 19 57

Farms in Raub 18 54

Farms in Bentong 1 3

Chinese (farm manager) 19 55

Malay (farm manager) 0 2

Male (farm manager) 17 52

Female (farm manager) 2 5

Average number of employed farm workers 3.3 1.2

Total farm land size (ha) 7.4 3.9

Total durian farm land size (ha) 5.7 3.7

Average monthly salary to workers (US$)a 346 308

Average total farm expenditure of 2012 (US$)a 17,862 6,172

The number of farms whose data of durian

produce of 2012 is available 18 47

Average farm durian produce of 2012 (ton) 25.2b 17.4b

Average durian produce per hectare (ton/ha) 4.4b 3.9b

a1USD was approximately 3.2 Malaysian Ringgit (MYR) at the time of the research.bThe results refer only to the farms whose data of annual durian produce are available.

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applying for it. Of the 20 uncertified farm managers with some knowledge of MyGAP, 8

managers (40%) pointed to the lack of tangible benefits from MyGAP certification as the

reason why they had not applied for the program4)

. Even though getting a GAP certification

has become the necessary condition for farmers to export their produce, simply holding it

does not guarantee their feasibility to do so in the sense that other conditions need to be met.

Fruit quality (e.g., flavour, size, appearance, etc.) is the main consideration of middlemen in

sending farmers’ produce to the export market.

Farmers’ understanding of the basic GAP concept

A critical issue in GAP is related to the extent to which certified farm managers do

understand the basic concept and purposes of GAP. The 19 certified farm managers

interviewed correctly pointed to food safety as the main goal of the MyGAP policy. This

result is in contrast to the findings of Amekawa (2013b) on Q-GAP, where over half of the

interviewed 64 certified pomelo farmers failed to identify the policy objective. This

discrepancy may be largely due to the contextual difference between the countries in terms of

the farm recruitment process for public GAP certification. In Thailand, the officially targeted

clusters of small-scale farmers who belong to a producer group are collectively promoted for

registration and provided education and training for the GAP program. Although the

decisions to participate are still at the hands of the individuals, the opportunities for them to

get access to information and resources could be significantly larger than those without a

membership of any producer group. This approach seems to solicit a situation where there are

farmers, especially old and less educated, who participate in the program rather passively,

failing to understand or remember what they have been involved in (Amekawa, 2013b). By

contrast, in Malaysia where the number of small-scale farms nationwide is much less and the

proportion of those who belong to a producer group is also much less than in Thailand, the

farm recruitment process is largely individually-based, and as such, there is not much

organizational mechanism for group-led certification. Due to the fact that decisions to

participate in the MyGAP program rely significantly on individual farmers, they tend to learn

as much as they could, try to comprehend the fundamental concept, and embrace its

significance.

Perceived benefits and shortcomings

Perceptions of certified producers about the benefits and shortcomings of MyGAP

certification on their farm operation may affect their decisions about reapplication in the next

round. Based on the questionnaire response, 10 farm managers (53%) said there are no

benefits from obtaining MyGAP certification, followed by 4 managers (21%) who pointed to

the acquired export opportunities as a merit of gaining MyGAP certification. Those farms

who used to sell durians to overseas markets before GAP certification was officially required

for export tend to undervalue the export opportunities acquired through certification.

However, those who began to seek exporting of durians after the certification requirements

4 In addition, 6 farms (30%) have not applied for MyGAP because they did not know sufficiently about it, and 4 farms (20%) attributed their

lack of application to the relatively small size of their durian orchard for export.

181

were put in place seem more likely to appreciate a GAP certification as beneficial for their

economic goals.

A more specific question on the perceived economic advantages of MyGAP certification

was asked to all the interviewed certified farm managers; it was as to whether they consider

themselves to have become economically more advantaged, remained the same, or less

advantaged after obtaining certification. Sixteen certified farm managers (86%) said their

economic status had remained the same. Eight of them attributed the view to the farm gate

price exhibiting no changes after they received certification. There were only 3 managers

(16%) who pointed to increased economic advantages. They considered the advantages to be

related to the export opportunities opened for them via gaining MyGAP certification.

It is interesting to note that the perceived shortcomings of participation in the MyGAP

program involved, among others, the burden of complying with certification requirements

pointed out by 6 farm managers (32%), followed by 5 managers (26%) who referred to

complex management procedures as such. These results appear to be consistent with the

results on the question of the most difficult thing to do in attempting to obtain MyGAP

certification. Five farm managers (26%) pointed to the difficulty in complying with the

requirements for pesticide control and/or passing pesticide residue sample analysis. While

seemingly not very significant, this result may suggest an important distinction from

Amekawa’s (2013b) study of Q-GAP, which found that Q-GAP certified farmers felt

virtually no pressure to change their pesticide practices due to its limited levels of required

stringency in compliance. MyGAP regulations, on the other hand, would have some

measureable effects on changing their behaviours related to pesticide use and control.

Another 5 managers referred to the difficulty in following tedious documentation

requirements in application and record keeping. A few even confessed regrets in having

applied for MyGAP because of the allegedly complicated application procedures required

relative to the tangible benefits they have obtained from certification.

DoA support for compliance

Regardless of whether durian producers have applied for MyGAP based on their self-

determination or encouragement by others, they need external training to gain an

understanding of compliance requirements and acquire necessary skills. Fourteen certified

(74%) and 4 uncertified farms (7%) received some kind of training provided by the DoA.

With regard to training on the use and handling of pesticides, 9 certified (47%) and 6

uncertified farm managers (11%) responded that they had received training through the DoA.

These results indicate that over half of certified farm managers have received neither

pesticide training nor have about a quarter of them had any MyGAP training through the

DoA. Although the DoA is the main enforcer of MyGAP regulations related to pesticide use,

many farm managers were self-reliant on how to abide by the rules and regulations related to

certification and pesticide use.

Farmers must know which pesticides are legal and which are not, given that illegal

pesticides are readily available at the local market. All the legal pesticides are registered

under the Pesticide Act, a law in Malaysia which was first introduced in 1974 and later

amended over time to control pesticide use. Eighteen certified (95%) and 44 uncertified farm

managers (77%) responded that they are aware of the types of pesticides officially registered

under the Pesticide Act. The majority of them (certified 80% and uncertified 53%) referred to

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agrochemical suppliers as the major source of such knowledge, reflecting the lack of training

they have received through the public sector. Farm managers were also asked whether they

sought out any advice on pesticide use from relevant authorities. Of the 13 certified (68%)

and 31 uncertified farm managers (57%) who answered that they did, 12 certified (93%) and

all the uncertified managers (100%) replied that pesticide suppliers are the main agents they

seek advice from. Only 1 certified farm manager and none of the uncertified managers sought

advice from the DoA.

Record keeping is a requirement of compliance for MyGAP certification. In the case of

GlobalGAP, farms applying for certification are most likely to fail if record keeping has not

been practiced properly even when all the other requirements have been met. This is not the

case with MyGAP where record keeping comprises only part of many compliance criteria

upon which the decisions of the DoA for certification are to be made. Asked about their daily

record keeping habits, 11 certified farm managers (58%) said they always keep records while

there are 3 managers (16%) who said they never do it. As expected, the majority of

uncertified farm managers (77%) never keep record and there are only 7 uncertified managers

(12%) who said they always maintain some form of record keeping. While there is limited

evidence available, this situation of record keeping of MyGAP certified farms is much better

than the case of Amekawa’s (2013b) study of Q-GAP, where most of the interviewed 64

certified pomelo farmers ceased to keep records after receiving certification. Farmers’

awareness of the importance of record keeping may be significantly different between

Malaysia and Thailand. Many Thai farmers certified based on group solicitation may not

readily understand what the Q-GAP policy is all about whereas most Malaysian farmers

certified on the individual basis may understand the basic principles and requirements of

MyGAP well.

Pesticide use

Pesticide use and handling practices comprise an important component of MyGAP as a

food safety standard. In the MyGAP code of practice, about 30% of control points are

directly relevant to synthetic pesticides. In case those of indirect relevance are included, more

or less 50% of control points become relevant, with the control category of Crop Protection

comprising the majority of 44 control points related to the use of synthetic pesticides (The

Department of Agriculture, Malaysia, 2005).

In the area under study, there are 3 kinds of synthetic pesticides that durian farmers were

using: insecticide, fungicide, and herbicide. Not all of the interviewed farms used the 3 kinds

altogether but around 40% of farms used either 1 or 2 kinds of synthetic pesticides (Table 3).

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Table 3. Pesticide use of respondent farms

To test whether MyGAP helps reduce the amount of pesticide use, comparisons of

certified and uncertified farms were made in terms of the annual quantity of active chemical

ingredients contained in and used for each of the 3 pesticide types, respectively5)

. Of the

farms who provided available information, uncertified farms were found to use 4.0 times

more insecticide than certified farms (p < 0.05). As for synthetic herbicide use, of the farms

who provided available information, uncertified farms were using 11 times more than

certified farms (p < 0.05). Synthetic fungicide tended to be applied to the trees with fungal

infections as needed. Of the farms who provided available information, the average amount

of fungicide used by uncertified farms was much less than the case of insecticide and

herbicide, with certified farms showing a slightly smaller amount of use than uncertified

farms. The seemingly tiny difference in the use of fungicide is indeed statistically significant

(p < 0.1).

Despite the incompleteness of the obtained data, in the comparison of the 11 certified

farms and the 36 uncertified farms whose data are available for all the 3 pesticide types,

uncertified farms are found to use nearly 4.0 times more aggregated amount of pesticides

than certified farms (p < 0.01). Despite with the limitations in the sample size for this study,

these findings appear to be distinctive from the aforementioned study by Schreinemachers et

al. (2012), which identified no statistically significant differences (p > 0.1) in synthetic

pesticide use between Q-GAP certified and uncertified FFV growers in Chiang Mai Province

in Thailand.

5 In these cases, the farms who do not use any kind of pesticides were included in the analysis with a ‘zero’ value given to the amount of a pesticide used in terms of active chemical ingredients. The active ingredients identified in the use on surveyed farms include: Abamectin,

Beta-cyfluthrin, Carbaryl, Chlorpyrifos, Cypermethrin, Deltamethrin, Diafenthiuron, Dimethoate, Fenthion, Imidacloprid, Lambda-

cyhalothrin, Malathion, Monocrotophos, and White Oil for insecticide; Benomyl, Difenoconazole, Fosetyl-aluminium, Maneb, Metalaxyl-m, Methomyl, Metiram, Propineb, and Triforine for fungicide; Diuron, Glufosinate-ammonium, Glyphosate isopropylamine, Paraquat

dichloride, and Phosphorus acid for herbicide.

Type of pesticide used Certified (19 farms) Uncertified (57 farms)

Insect ic ide

The number of farms whose data are available 17 (84%) 48 (84%)

The number of farms who use insecticide 17 (100%d) 45 (94%d)

Annual amount of active ingredients (a.i.) per hectare (kg/ha) 1.01b 4.04

Fungic ide


The number of farms who use fungicide 4 (27%d) 10 (19%d)

Annual amount of a.i. per hectare (kg/ha) 0.23c 0.25

Herbic ide


The number of farms who use herbicide 13 (93%d) 32 (65%d)

Annual amount of a.i. per hectare (kg/ha) 0.69b 7.26

All the Pest ic ides (Insecticide + Fungicide + Herbicide)


The number of farms who use at least one type of pesticide 9 (82%d) 34 (94%d)

Annual amount of a.i. per hectare (kg/ha) 1.42a 11.37a p < 0.01, bp < 0.05, cp < 0.10; NS not significant at 0.01.dThe percentage refers to the number of farms who use a particular pesticide in question divided by the number of farms whose data are available.

184

While certified farms, overall, are found to use significantly less synthetic pesticides

than uncertified farms, the former seems to be more convinced of the safety in pesticide use

than the latter. All of them expressed the belief that, if properly managed, pesticides are

harmless to pesticide applicators, consumers, and the environment. Uncertified farms appear

to be more doubtful. Twelve of them (21%) mentioned that, even if properly managed, some

levels of harm are unavoidable to pesticide applicators. Nine uncertified farms (16%)

expressed the possibility of any harm to consumers even in conducting appropriate pesticide

management, and 4 uncertified farms (7%) pointed to such a risk with respect to the

environment, respectively.

Non-synthetic, alternative pest management practices could be employed as part of an

integrated pest management strategy for the reduction of pesticide use and associated social

and environmental costs. There are 6 certified (32%) and 6 uncertified farmers (11%) who

use one or more pest management methods other than synthetic pesticides, with a statistically

significant difference (p < 0.05) (Table 4). Special mention needs to be made of alternative

weed management methods. While there are 16 certified farmers (84%) who use synthetic

herbicide with a much smaller average amount than uncertified farms, there is only 1 farmer

who relies on an alternative weed management method: mechanical weed cutter. On the part

of uncertified farmers, there is only 1 farmer who substitutes herbicide use by a non-synthetic

method: the manual removal of weeds. This evidence suggests that, while using little or no

herbicide, the majority of certified farmers let weeds grow on their own without caring much

about the potential adverse ecological consequences such as weed-tree competition over soil

nutrients. Many certified durian farmers responded that they are not concerned about weed

growth very much since they consider that its negative effects on tree and fruit growth are

negligible in the case of durian farming in the region. Further, some explained that many

farms are just too large for the amount of labor required for manual or mechanical weeding

methods, given that weeds regularly grow in weeks after cutting them. Such methods are

considered less efficient than the power of herbicide leading to the elimination of weeds for

several months to come.

185

Table 4. Alternative pest management by respondent farms

Pesticide handling

Pesticide handling practices of certified and uncertified farms were examined comparatively

in terms of selected items covered and not covered in MyGAP guidelines (Table 5). Of the 7

items covered in MyGAP guidelines, all of them present no statistically significant

differences (p > 0.1) except for item 7 on the possession of a pesticide storage (p < 0.01). It

should be noted that there is one farm who does not have a storage that specialises in the

housing of pesticides even though the farm has been MyGAP certified. With regard to the 7

items that are not covered in MyGAP guidelines, 4 items showed a statistically significant

difference (items 9 and 10 for p < 0.05 and items 11 and 12 for p < 0.01), with all of them in

favour of certified farms. In item 8 (the observance of pesticide labels for pre-harvest

intervals), the ratio of farms following the practice appears low for both certified and

uncertified farms. The results turn out to be misleading because the majority of farms follow

their own rules for pre-harvest intervals. Three certified (16%) and 3 uncertified farms (5%)

mentioned, however, that they use pesticides as needed while not following any pre-harvest

intervals.

Certified (19 farms) Uncertified (57 farms)

The number of farms who adopt alternative

pest management 6a (32%) 6 (11%)

The number of certified farms who use:

Rodent trap 2 0

Biological control (birds) 2 0

Cutting weeds 1 0

Burning litters 1 0

Shot gun 0 1

Wire fence 0 1

Mesh wire trap 0 1

Cats catching rats 0 1

Smoke release to scare pests 0 1

Biological control (lizards) 0 1

a p < 0.01, bp < 0.05, cp < 0.10; NS not significant at 0.01.

186

Table 5. Pesticide handling of respondent farms

Overall, pesticide handling practices of certified farms are better than uncertified farms

with 5 of the total 14 items exhibiting a statistically significant difference (p < 0.05 or p <

0.01). Although a direct comparison may not be appropriate, it should be noted that in the

aforementioned Schreinemachers et al.’s study (2012), no significant differences in pesticide

handling were found between certified and uncertified farms for 7 items6.

CONCLUSIONS

This study on Malaysia’s MyGAP standard has investigated the relationship of safety

assurance and farm participation by examining the perceptions of relevant durian farmers and

their pesticide use practices in the state of Pahang, Peninsular Malaysia. It was found that the

19 certified farm managers who were interviewed understand the fundamental rationale of

MyGAP. The result is clearly different from the finding of Amekawa (2013b) that over half

of the interviewed 64 certified pomelo farms did not show an understanding of the food

safety goal of Q-GAP. This difference would be related to, among others, the institutional

dissimilarity between the 2 programs: the application of small-scale farms for Q-GAP is

largely group-led, giving advantage to the farms affiliated with a producer association. The

majority of FFV farms in Malaysia are independent from producer groups and their

6 The 7 items in Schreinemachers et al. (2012) include: 1. Use pesticides in a preventive way (regular spraying); 2. Follow product labeling

to decide on dosage to use; 3. Take temperature or radiation into account when spraying; 4. Take wind speed and/or direction into account when spraying; 5. Cover mouth when spraying; 6. Cover arms and legs when spraying; and 7. Take a shower and wash clothes after

spraying (p. 524).

Certified (19 farms) Uncertified (57 farms) t-test

Items covered in MyGAP guidelines

1. Change clothes after spraying pesticides 18 (95%) 50 (88%) NS

2. Wear long-sleeved shirt for spraying 18 (95%) 50 (88%) NS

3. Wear long-sleeved pant for spraying 19 (100%) 55 (96%) NS

4. Wear mask for spraying 19 (100%) 56 (98%) NS

5. Take care of wind direction while spraying 18 (95%) 49 (86%) NS

6. Follow product label to decide on the dosage 10 (53%) 33 (58%) NS

7. Have a pesticide storage that does not store 18a (95%) 40 (77%)

other things but pesticides

Iterms not covered in MyGAP guidelines

8. Strictly follow the pre-harvest intervals 5 (26%) 21 (37%) NS

as prescribed on pesticide labels

9. Smoke while spraying pesticides 0b (0%) 4 (7%)

10. Eat anything while spraying pesticides 0b (0%) 4 (7%)

11. Drink anything while spraying pesticides 0a (0%) 14 (25%)

12. Take shower within one hour after spraying 17 (89%) 44 (77%) NS

13. Change clothes after spraying and as soon as 19a (100%) 45 (79%)

arriving at home

14. Wash clothes used during spraying together 10 (53%) 25 (44%) NS

with clothes not used for spraying

a p < 0.01, bp < 0.05, cp < 0.10; NS not significant at 0.01.

187

applications for MyGAP tend to be individually-based, thus likely to invite the applications

of farms who are sufficiently conscious of the basic intent of the program.

The majority of certified farm managers decided to apply for the MyGAP program

because they wished to either maintain or newly open their durian export opportunities

through obtaining MyGAP certification, given that a certification is the officially required

condition for the exportability of their produce. Farm managers who used to export durians

before a GAP certification became the official prerequisite for export rather take for granted

the export opportunities acquired through a MyGAP certification and thus perceive a

certification as not very beneficial. Those who began to seek durian export after the

certification requirements were put into effect tend to see a certification as more beneficial

for their economic objectives.

Regarding pesticide use practices, certified farms are found to use significantly smaller

annual amount of pesticides for each kind (insecticide, fungicide, and herbicide) as well as

for all the 3 kinds combined. This finding is in contrast to Schreinemachers et al.’s (2012)

study of Thailand’s Q-GAP, which found no statistically significant differences in the annual

amount of pesticide use between certified and uncertified FFV farms in Northern Thailand.

However, this is within the scope of our expectations as it was hypothesized that MyGAP

certified farms would exhibit significantly better results than uncertified farms due to its

much higher level of compliance required for certification than the case of Q-GAP. Coupled

with the small sample size and limited available data obtained in this study, however, the

obtained results should not take an outright acceptance. More studies of MyGAP and other

public GAP standards that are implemented in ASEAN countries are definitely needed to

gain national- and ASEAN regional-level insights into the official balancing act between food

quality assurance and small-scale farm participation.

ACKNOWLEDGEMENT

The authors are deeply thankful for the DoA Malaysia, DoA Raub and DoA Bentong for

allowing us entry and investigation in the districts under study. We also acknowledge the

financial support of the University of Malaya through the University of Malaya Research

Grant (Project RG186-12SUS).

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191

FFTC-KU 2015 Organizing Committee

FFTC

Dr. Takashi Nagai

Deputy Director, FFTC Dr. Wan-Tien Tsai

Agricultural Specialist, FFTC

Mr. Ronald Mangubat

Information Officer, FFTC Ms. Claire Fang

Secretary, FFTC

192

FFTC-KU 2015 Organizing Committee

Kasetsart University

Dr. Warapa Mahakarnchanakul

Department of Food Science and

Technology, Kasetsart University

Head of the project

Dr. Kanithaporn Vangnai (Jah)



Secretary

Dr.Kullanart Tongkhao



Committee

Dr.Kriskamol Na-Jom



Committee

193

Sponsor

Kasetsart University Food Innovation Research and Services in Thailand

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