IT final report

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Universiti Tunku Abdul Rahman

Faculty of Science

Bachelor of Science (HONS) Biochemistry

Year Two Semester Two

UDEE 2306 INDUSTRIAL TRAINING

REPORT

NAME : CHEW HUI SIN

ID NO : 0908196

COMPANY : BIOFACT LIFE SDN BHD

ADDRESS : LOT 5094, KAWASAN PERINDUSTRIAN

PARIT JAMIL, 84150 PARIT JAWA,

MUAR, JOHOR

SUPERVISOR (COMPANY) : Mr NG HONG SENG

SUPERVISING LECTURER : DR KHOO KONG SOO

DATE OF SUBMITION : 13 JANUARY 2010


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CHEW HUI SIN 0908196

UDEE 2306 INDUSTRIAL TRANING

2

Table of Contents

Table of Contents ....................................................................................... 2

Acknowledgement ..................................................................................... 3

Abstract .................................................................................................. 4

Introduction ............................................................................................... 5

Goal of Industrial Traning ......................................................................... 5

Scope Of Training ................................................................................. 5

Company Background ............................................................................... 6

Mission& Vision ....................................................................................... 7

Award and Accreditation ........................................................................ 7

Product ................................................................................................... 10

Work-Based Learning Experience .......................................................... 12

Learning Outcomes ................................................................................. 19

Microbiology Lab .................................................................................... 19

Chemical Lab ...................................................................................... 22

Conclusion ............................................................................................... 23

References ............................................................................................... 24

Appendix ................................................................................................... 25

Bi-weekly reports ..................................................................................... 25

Project Assigned ................................................................................... 26


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ACKNOWLEDGEMENT

I feel so glad and appreciated with my University, Universiti Tunku Abdul

Rahman, give me an opportunity to go through this industrial training. This program

helps us to get use with the truly working environment prior our graduation to increase

our competitive force in the real world. I would like to thanks Dr Khoo Kong Soo,

lecturer supervising me in Utar that willing to guide me and giving me direction from

time to time during the industrial training.

I would like to thanks my company, Biofact Life Sdn Bhd that providing me an

excellent working environment for me learn. Not to forget the science executive of our

company, Mr. Eston Lee, which willing to hire me as a trainee and also assigned me to

different department to handle different kinds of tasks.

Last but not least, both of my supervisors which are Mr Ng Hong Seng and also

Miss Lee Chia Yen. I am so appreciate with their encouragement to share their working

experience with me. By the way, they also assisting me in complete the project offering

to me. I do learn a lot of knowledge in their department.


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ABSTRACT

Industrial training is credited course programmed that compulsory to satisfy as a

degree course work requirement for graduation. As a biochemistry student, all of us have

to undergo the 12 weeks industrial training from 4 october until 24 December. This report

will cover the essential aspects of my industrial training experience. From the goal of

which I would want to achieve from this experience to the learning and training grounds

which I have undertaken. This report will introduce the company profile first. Secondly, I

would be mentioned my learning experience within this 12 weeks of working period.

Thirdly, I will include what I had gained and applied in this report. Last but not least, the

bi-weekly report and sample of work done would be included as appendices in the end of

the report.


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INTRODUCTION

Goal of Industrial Training

The goal of this industrial training is to assist the students by exposing them to the real

working world. Within twelve weeks, students are exposed to the industrial so when it is time for

them to venture into work life it would not come as a shock.

Another goal is to gain experience in real working environment. This is so that students

are able to learn work ethics, take on and handle responsibilities and meet deadlines efficiently.

This industrial training also enhances and assists students to be aware of their strengths

and weaknesses and to grow and improve themselves from here on. With this realization, students

are able to build confidence in their capabilities and use theories learnt in classes in real life

situations.

Scope of Training

In the beginning of the internship in Biofact Life, I was assigned to the chemical

laboratory. Under the guidance of Miss Lee Chia Yen, our company chemist, I learn to handle the

main instruments in the lab. I was getting an experience on how to become a good assistant in the

lab to assist my supervisor to complete her daily routine works.

Besides, I also get a chance to learn from the microbiologist in our company. Our

company microbiologist, Mr Ng Hong Seng is also quality assurance in our company. Thus,

besides working as a trainee in the microbiology lab, he also introduces me on how a QA

department operate in the company. He also gives an opportunity for me to become a one day

microbiologist in the company in order to let me experience the real working environment more

deeply.


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My company also assigned me a project regarding to cordycepins extraction. Thus, I

planned and carried out difference type of experiment in order to complete the project. And Mr

Ng Hong Seng and Miss Lee Chia Yen will assist and guide me throughout the whole project.

Company Background

Biofact Life Sdn Bhd was incorporate in Malaysia under the Company Act 1965 on 1st

February 2005. Biofact Life Sdn Bhd specialises in the production of cordyceps by laboratory

cultivation. The company is one of the pioneer biotechnology companies in the world to employ

the latest biotechnology techniques in cordyceps cultivation at reduced cost and improved quality.

They culture the cordyceps under low temperature and oxygen conditions. They designed

environment that free from chemical pollution as well as all the yeast, mould and bacterial

contamination.

BioFact Life encompassed a seed-to-shelf approach as the company is fully committed to

extensive research and development in the production, manufacturing and marketing of their

cordyceps. GMP and ISO 22000:2005-certified are practise in the manufacturing division. Thus,

the quality, safety and efficacy to manufacture the product in the form if capsule, powder, tablet,

pill, liquid, and ointment are maintained.


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Vision

Quality for a quality life

Mission

y To become a key global player in the mainstream of natural healthcare markets

through vertical integration and strategic globalization.

y To develop, manufacture and promote quality natural healthcare products for

mankind.

y To keep abreast with the latest developments in natural healthcare products.

y To ensure continuous improvement in the standard of our products and services so

that our company, employees, business partners and stakeholders will benefit and

prosper together.

Award and Accreditation received by BioFact Life

Golden Bull Award 2009

BioFact Life Sdn Bhd has been awarded with the Golden Bull Award 2009, which is most

representative annual business award that accredits and honours the best efforts and achievements

if SMEs in Malaysia.


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SMEs Fast Moving Companies 2009In 2009, Biofact Life also received recognition from the SME Fast Moving Companies. The

SME100 is an Annual Recognition Award arganized by SMe & Entrepreneurship Magazine for

recognizing the achievements of SMEs in Malaysia.

ISO 22000:2005

Biofact Life Snd Bhd demonstrating their commitment to ensuring the higest standard food safety

management also received ISO 22000:2005 certified in the first quarter of 2009. Through this

certification, they ensure that the mist effective food safety systems are established operated and

updated within the framework of structured management sysyatem and incorporated into overall

management activities of BioFact Life.

The Malaysia Book of Records

BioFact Life has been accreditated by the Malaysia Book Of Records as the First Malaysian

company that successfully cultivate cordyceps in a specially designed 100K clean room where the

room conditions are regulated and controlled according to the different growth stage of

cordyceps.


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Halal Product Certified

BioFact Life also obtained a Halal Certification from the Department of The advancement of

Islam Malaysia (JAKIM).

Good Manufacturing Practice

BioFact Life is GMP- Certified. The company takes proactive steps to ensure that the products

are safe, pure and effective by enforcing a quality approach in the manufacturing, processing and

packaging processes.

Bioinno Awards 2008

BioFact Life also awarded the Gold Winner in Bioinno Award Competition organized by the

Ministry of Science, Technology and Innovation (MOSTI).


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Product Manufactured by BioFact Life

The pharmaceutical products manufactured by BioFact Life are basically added up with

cordyceps they cultivated. Those are the hot seller product in our company:-


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Work-based Learning Experience

Cordyceps Sinesis, which is known to the Chinese as Dong Cong Xia Cao and

to Japanese as Tochukuso was long been used in medicine.(John et al.,2004) The native

cordyceps are grown on insect such like worm. The insects body play role in providing

nutrient for the cordyceps to growth. The cordyceps cultured in our company are using

rice as the substrate for cordyceps. After the cordyceps being harvested, the substrate is

remained. But the substrates do absorb some unique ingredient found in cordyceps such

like cordycepins. My company would like to extract the cordycepins found in substrate

for others usage in the industrial.

So, my company assigned me the project to set the procedure for the cordycepins

extraction. Firstly, I planned the overall extraction process. The process is as showed in

the flow chart below;-

In the first step, I plan to using heat to release the cordycepins found in cordyceps

substrate. Then I will filter out the residual. The extract might full with different large

compound or molecule like polysaccharides from the substrate. So, I try to using heat to

Incubatethe

substrate

Filter Boil the

extraction

Filter

Concentratethe

extraction

Liquid Extract of

Cordycepins

Autoclavethe

Extract


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break down those molecules that consider as the impurities in the extract by using heat. The

impurities is then filter out. We will set the minimum concentration of cordycepins should

presence in the extract to 300ppm. If we could not get the concentration we desired directly

from the process, concentration method is needed to concentrate the extraction. At the end

we will autoclave the final extract to kill the bacterial thus contamination is avoided.

There are differences factors that may affect the concentration of cordycepins in the

extract. The factors I consider about are:-

y Ratio of substrate to water

y

Temperature

y Incubation time

y Concentration method using

y Pre-treatment involve

Differences types of experiment are carried out to determine whether those factors can affect

the concentration of cordycepins during the extraction.

Simple materials and methods are using throughout the project. Apparatus such

like beaker, Schott bottle, Bunsen burner, spatula, retort stand, Whatmanfilter paper, filter

bag, filter pump and thermometer are using in the few experiments. Besides, HPLC,

incubator and autoclaving machine are also needed in the project.

The first experiment I conducted is to find out which ratio of water to substrate can actually

work the best in the releasing of cordycepins in water. Firstly, a specific amount of substrate

is weighted and placed in a Schott bottle. The Schott bottle is then fill in with water before

place in a 60 C water bath to incubate for 2 hours. The residual is filtered out by using filter

bag. The extract is then sent to autoclave at 122 C for 15 minutes to break down the


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polysaccharides and also all the impurities in the extract. A thick precipitate will usually

found in the extract, hence, we need filter paper to filter the precipitate out. I will autoclave

the extract at 122C for 15 minutes to avoid contamination occurs. The final extract is then

sending to chemical for HPLC test. The variety of ratio and the result is stated in the table

below:-

As the conclusion, the ratio of substrate to water 1:7 can work the best in the

extraction process. This ratio showed us the closest figure between the actual and expected

concentration of cordycepins in the extraction. This means that the most cordycepin found in

the substrate were extracted out and only few amounts of cordycepins are remaining in the

substrate or residual.

The second experiment carried out is about the optimum temperature of

cordycepins extraction. There are data showing that the melting point of the cordycepins is

225 C. Therefore, I expect that the concentration of the cordycepins in the extract will

Substrate :

water

Substrate

weight (g)

Volume of

water (ml)

Cordycepin

being tested

(ppm)

Expected

concentration

of cordycepins

Effectivenes

(%)

1:1 250 250 762.347 762.347 100.00

1:4 100 400 132.147 190.587 69.33

1:5 60 440 113.628 152.469 74.52

1:6 70 430 99.252 127.058 78.11

1:7 80 420 94.973 108.907 87.2

1:9 50 450 68.520 84.705 80.88


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increase as the incubation time increase. The same steps in the first experimrnt are carried

out. But the temperature using are differences.

The result is as tabulate in the table above.

From the table above, we can conclude that the melting point is not as the same as

the temperature which cordycepins can resist. Its only shown that the physical change of

cordycepins change from solid to liquid. The best temperature get is 60 C. The hypothesis I

made are rejected as the concentration of cordycepins in the extract decrease when the we

set the extraction temperature into 75 C. while cordycepin also cannot be extracted well at l

room temperature.

Then, the experiment is followed by determine the best incubation time for the

extract. Same experiment are carried out the only different is the time taken for incubation.

We use 1, 2 and 3 hours respectively in this experiment. I do a similar hypothesis as the

previous experiment in this experiment. I thought that the concentration of cordycepins

might increase as the time of incubation increase at a constant temperature. I found out that

my expectation wrong once I received the chemical result. The result is as shown as below:-

Temperature (C) Concentration of cordycepin

obtained (ppm)

Percentage water

remained (%)

25 70.68 72.7

50 87.45 63.6

60 94.97 63.6

75 81.66 36.4

100 - 0


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As a conclusion, long period of incubation doesnt showing the positive result all

the time. The concentration of cordycepins shown us a slightly increase trend in the first two

hours. Unfortunately, the trend doesnt hold at the 3 hours of incubation. I belive that the

long period of incubation could not increase the concentration of cordycepins but only

degradation after long period expose to heat. For this reason, the cordycepins amount

decrease as the time increase. Consequently, the best incubation time I would suggest is

around 1-2 hours.

As I stated before, the concentration method is need to concentrate the extract in to

our minimum requirement, 300ppm. So, I had chosen 2 methods in this experiment to

concentrate our extract. I planned to use water bath and direct heating to concentrate the

extract. Before the experiment, the formula M1V1=M2V2 are using to calculate out the

expected concentration. The apparatus are showed as the picture below:-

Time ,t (hr) Concentration of cordycepin

obtained (ppm)

Percentage of water

remained,%

1 90.50 63.63

2 94.97 63.63

3 88.94 45.45


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The detail procedure is written in the report attach to the appendix. And the results are tabulated

in the table below:-

As the conclusion, water bath is a better concentration method as it can remain the quantity

as well as the quality of the extract after the concentrate process. But water bath is not consider as

an efficient method as it using a very long period to evaporate only 40 ml of water in this

experiment. Hence, it is not so practical in the field of industrial as compared with direct heating

method.

Since the releasing rate of the cordycepins from the substrate has not achieve the

maximum concentration we predict. Thus, I try to find a way to increase the releasing of

cordycepins in the extract. I try to change the form or state of the substrate through pre-treatment.

Method Time taken Final volume

(ml)

Expected concentration of

cordycepins (ppm)

Actual concentration of

cordycepin (ppm)

Water Bath 2hrs40mins 7 646.43 655.00Direct Heating 38minutes 10 452.50 365.12

Direct Heating Water Bath


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I compared the concentration of autoclaved substrate and powder form of the substrate with

normal substrate. The chemical results are as shown below:-

Type of pre-treatment

Cordycepin being tested,

(ppm)

autoclaving 576.67

powdery 762.37

Without pre-treatment 657.63

It is not surprise that the powder form substrate give us the highest concentration of

cordycepins, as the size of the substrate decrease, the surface area increase, hence, releasing rate

of cordycepins increase. But the result of the autoclaved substrate showing us that the high

temperature during the autoclaving may broke down the structure of cordycepins in the substrate

as well as in the extract. So, the use of autoclaved machine should lesser in order to maintain the

quantity of the cordycepins.


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LEARNING OUTCOME

Different type of skill and knowledge I had applied and gained during the internship. The

basic knowledge that taught in our degree and also foundation helps me to perform well in the

working environment. Besides, those basic also help me to understand all the theories that are

new to me such like the HPLC, AAS and also microbiology theory and techniques that still not

yet learn in our syllabus.

MICROBIOLOGY LAB

I learn quite a lot of microbiology techniques in the microbiology. The first technique I

learn in microbiologist lab is how to serial dilution the sample product. Usually we will add one

ml of original sample are taken and add it to 90 ml of Tryptic Soy Broth (TSB), to give 10-1

dilution of original sample are taken and add it to 9ml of TSB. Similarly we can prepare 1:100,

1:1000, 1:10000 and so on to dilution of the original. Finally, one ml of any dilution is added to

sterile petri dish to which are added with sterile, cool, molten agar medium. The dishes are then

incubating at a suitable temperature within some period. After the incubation, colonies of each

microbe grow in the dish. The number of colonies of each kind is counted. The number is then

multiplied by the dilution factor to find the total number of cells per ml of the original sample.

Usually we will use the formula below to count:-

CFU (colony formed per unit)/ ml or g =

Besides, I also learn about different method of pure culture that usually use in routine

laboratory work. Before learning about the pure culture method, I learned about how to prepare

and store different type of medium and agar such like TSB, Sabouraud Dextrose Agar (SDA),

Baird Parker (BP) Agar and Levine Eosin Methylene Blue (LEMB) agar, and also the uses of

those medium and agar. Pour plate method and streak plate are usually use in our company. In

pour plate method, the sample is diluted in to a broth. The agar medium is maintained in molten

Colonies counted (average) Dilution Factor

actual volume of sample in dish, ml


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state at 45 C. One ml of each dilution is added to each sterile Petri dish to which is then poured

9ml of sterile, cool and molten agar medium. The contents are thoroughly mixed and allowed to

solidify. The dish is then incubating in a suitable temperature. In streak plate method, a small

amount of a sample is transferred into the surface of a suitable, solid agar medium either by loop

or transfer needle. This is then streaked in such a way as to provide successive dilution and

ultimately to have well isolated colonies. Streaking may be done in any do the ways. The sample

in each case will becomes progressively diluted and at the end of streak one would expect the

well isolated colonies. The result is of streak plate I did is shown below:-

Staphylococcus on Baird- Parker agar

Black shinny colony formed.

(Merck, 2005)

E.Coli on LEMB

Metallic shinny sheen in reflected light

(Merck, 2005)


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I also learn about carry out conformational test. We can confirm the presences of

staphylococcus aureus from coagulase-negative staphylococci by using Coagulase. By the way, I

also learn how to differentiate Grams Positive bacterial and Grams Negative Bacterial. Before

carry out the Gram stain, I try to prepare Bacterial Smears (Cappuccino&Sherman, 2004). The

procedure is basically as shown in the flow chart below:-

Then, the smear is flood with crystal violet. Wash it off after I minute by using tap water.

The smear is now purple-blue in color. Then, stain it with Grams Iodine for another 1 minutes.

The Grams Iodine is then washing off after 1 minute. The smear is then added with Decolorize

agent (95% ethyl alcohol) until showing only blue tinge. Gently wash the decolorized smear with

tap water and counterstain with safranin for 45 seconds. Lastly wash the smear with tap water and

blot dry with bibulous paper before examine under a microscope. Grams positive cell such likestaphylococcus have thick peptidoglycan layer, whereas the peptidoglycan layer in Grams

negative organism is much thinner and surrounded by outer lipid-containing layers. Grams

positive bacterial will shown blue color when we exam it under microscope. Grams negative

bacterial will counterstained by red safarnin, thus, pink color showed under a microscope.

Prepare a clean

glass slide

Drop 2 loopfuls of water on the

center of the glass slide and dilute

the culture placed.

Transfer the

culture to the

slide

Air dries for few minutesHeat Fixation


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CHEMICAL LAB

I applied the thing I learned in school life in chemical lab. I know how to prepare solution

from their stock solution. I know how to dilute the solution in to the concentration we need by

using volumetric flask.

Besides that, I also learned about the new equipment such like AAS, HPLC and also

Microwave digester in the lab. These 3 equipments play an important role in our chemical lab.

The learn how to operate the microwave digester and also how to clean it after use to maintain its

performances. The Microwave digested always deal with concentrate acid, must be very careful

when handling it.

Our company uses AAS (Atomic absorbance spectrometer) determine the concentration

of heavy meter found in the final product manufactured by our company to ensure that all of the

products are safe to be consume. AAS is an equipment special designed for determine the

concentration of different heavy metal in a sample. The presence of Cadmium, mercury,lead and

arsenic are determined in the product. Usually I will prepare the different concentration of

standard solutions which will use to calibrate the standard curve. The sample is only allowed to

enter the system for AAS test after the standard graph is plotted.

High- Performance Liquid Chromatography (HPLC), is widely use to separate and

determine species in a variety of organic, Inorganic and also biological material. (Leong, n.d) We

usually use it o determine the concentration of cordycepins and adenosine in our product as well

as the cordyceps we cultured. Varieties of detectors are found in the market. But BioFact Life

only uses UV detector and refraction detector in their daily routine works. I am so glad that I have

an experience on how to operate HPLC.


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CONCLUSION

As a conclusion, industrial training has introduced me the real working environment I

will face in the future. I have gained a lot of knowledge which I might not have an opportunity to

learn in book and class. For example, I learned how to communicate with people and how toadapt with the company environment and culture once I step out the University. I learned

different types of new skill which are very useful not only in my studies but also in the future

when I work. For example, have a chance to operate the equipment in chemical lab such like

AAS and HPLC give me advance knowledge. Through the industrial training, I have developed

new skill and also polished the skills that I am weak at.

In the twelve week of industrial training, I made new friends. I learn how to interact with

new people and how to cooperate with my colleague in the working area. From time to time, I

saw my weaknesses in communication with other and also how to express myself. My supervisor

and all the colleagues in the company have given me pieces of useful advices for me to improve

myself. Such like my communicating skill and also presentation skill as I am very shy when

needed to talk in front of new people. I will try my best to improve all the weaknesses.

I believe that the unforgettable experience I gained in BioFact Life has prepared me well

for my employment. Industrial training also helps me to identify my future career. I become more

confidence and skillful after the industrial training. For me, working in laboratory is so

challenging and interesting. In my opinion, gain knowledge through the industrial training is

more interesting compare to University learning environment. I hope that I will have an

opportunity to having this kind of experience in the time remaining at UTAR.


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REFERENCES

1. Leong,L.K.(n.d).Chapter 3: HPLC[powerpoint slides].Unpublished manuscript, UESC

3254, University Of Tunku Abdul Rahman,Malaysia

2. John,C.H.,Phillip,C.,Megan,L.P.,Dinesh,P.(2004).Analysis of Quality And Techniques

For Hybridization Of Medicinal Fungus Cordyceps Sinesis (Berk.)Sacc.(Ascomycetes),1-

2.

3. J.G.Cappccino.,N.Sherman.,(2004)Microbiology A Laboratory Mannual(7th ed.) . San

Francisco: Benjamin Cummings.

4. Microbiology Manual (12

th

ed.).Germany: Merck.


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