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Herbal Medicines: Indigenous, Molecular Aspects, and Clinical Application

.J Faculty ofPhannacy, Universitas Padjadjaran ISBN 978-602-96121-1-0

I

Proceeding

The International Seminar and Expo on Jamu 2010 (ISEJ 2010) HHerbal Medicines: Indigeneous, Molecular Aspects, and Clinical Application"

Edited by: Faculty of Pharmacy, Universitas Padjadjaran

Published by: Faculty of Pharmacy, Universitas Padjadjaran

Copyright © 2010 by Faculty of Pharmacy. All rights of reproduction in any form reserved

ISBN 978-602-96121-1-0

911l111jlj'11!III~ \1~\I~ltl"

Foreword from Dean of Faculty of Pharmacy Universitas Padjadjaran

Dear Delegates,

On behalf of the Conference Committees, I would like to thanks for your participation in The International Seminar and Expo 2010, which take place from November 5th..6th 2010, at Bandung, Indonesia. This seminar of "Herbal Medicines: Indigenous, Molecular Aspects, and Clinical Application" offer a comprehensive understanding of utilization of herbal medicines from various aspects, including those associated with their regulation, traditional use, chemical analysis, biological activity, mechanism of action, and clinical application.

This proceeding is consist of approximately 34 papers. We thanks to The President of Universitas Padjadjaran, Prof. Dr. Ganjar Kumia, DEA.; The Dean of Faculty of Pharmacy of Universitas Padjadjaran, Prof. Dr. Anas Subamas, M.Sc., Apt.; and all the authors that participated in this conference for all their support and contribution in publishing this proceeding.

As the organizing committee, we greatly appreciate your participation in The International Seminar and Expo 2010. We look forward to meeting you and welcoming you again in our next meeting.

Bandung, November 5th 2010. Sincerely,

Prof. Dr. Anas Subamas, M.Sc., Apt. Dean of Faculty of Pharmacy Universitas Padjadjaran

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Content

Page Foreword

Coconut Water To Induction Multiplication Of Piper crocatum L. (Alfandi, R. Eviyati, Suskandini) ...................................................................................... ..

Comparison Of Antioxidant Activity And Total Phenolics Content Of Aqueous Extracts Of Labisia pumUa var. alata From Malaysia And Indonesia (Ade C. Iwansyah and Mashitah M. Yusoff) .......................................................................... 6

Antidiarrheal Activity Of Water Extracts Of Sam bang Getih Leaves (JIemigraphis alternata (Burm. F.) T. Anders) In Swiss Webster Mice (Puspa Sari Dewi, Afifah B. Sutjiatmo, Kusbudiantoro) ..................................... 11

Essential Oils Content Analysis And Anti-Inflammatory Activity Test From Extract Of Kaempferia galanga Linn. Rhizome (Aliya N Hasanah, Fikti Nazaruddin, Ellin Febrina, Ade Zuhrotun) .................................... 14

In Vitro Anti-Malarial Alkaloid And Flavonoid From The Erythrina Variegata (Leguminoseae) Plant (Tati Herlina, Unang Supratman, Anas Subamas, Supriyatna Sutardjo, Suseno Amien, and Hideo Hayashi) ...................................................................................... 20

Antibacterial Activity Of Residue Fraction Ethanolic Extract Of Phyllantus acidus (L.) Skeels Against Staphylococcus aureus And Eschericia coli Antibiotic Multiresistance (lka Trisharyanti Dian Kusumowati, Maryati, Prasetyo Adi Laksono) ............................... 25

Fingerprint Pattern Of Thin Layer Chromatography And High Performance Liquid Chromatography Of Evodia suaveolens Scheff Leaves Extract (Wiwiek Indriyati, Mutakin, Neily Zakiyah) ....................................................................... 32

In Vitro Determination Of Antioxidant Activity Of Extracts Of Mahkota Dewa, Temu Putih, Sambiloto And Keladi Tikus (Dyah Iswantini, Gustini Syahbirin and Dedy Irawan) ........................................................ 39

Determination Of Anti-Hyperuricemic Extract Of Celery, Sidaguri, And Tempuyung By In Vitro Method CDyah Iswantini, Min Rahminiwati and Dian Itkarullzzah) .............................................. 46

Cytotoxicity Test of Ocimum americanum L. Leaves Extract Using Brine Shrimp Lethality Bioassay (BSLB) (Sesilia Andriani Keban, Shirly Kumala) ............................................................................. 50

Interaction Of Andrographolide And 14-Alpha-Lipoyl Andrographolide (Ala) With Neuraminidase Of H5n1 Virus (Jutti Levita, Masripah, Driyanti Rahayu, Muchtaridi) ........................................................ 54

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iii

The phytochemistry study and antidiabetic activity of red katuk (Euphorbia cotinifolia L.) Leaves (Ria Mariani) ........................................................................................................................ 59

Effect Of Drying Process And Raw Material Grinding On Essential Oil Profile Of Zingiber Cassumunar Roxb. (Lia Marliani, Sukrasno, and As'ari Nawawi) ..................................................................... 68

Antioxidant Activity Of Joseph's Coat (Amaranth us tricolor L.) With Dpph Method (Nyi Mekar Saptarini, Dadan Suryasaputra, and lla Sajarotuddur) ..................................... 72

Chlorofom Fraction Of The Ethyl Acetate Extract Of The BarkOfKluwih (Artocarpus communis Park) Is More Cytotoxic On T47d Cells Than Hela Cells (Rosita Melannisa, Ratna Yuliani, and Muhtadi) ................................................................. 76

Antiseptic Effectiveness Studies OfAgeratum conyzoides, L (Soraya Ratnawulan Mita, Ellin Febrina , Anggraeni Wulandari) ....................................... 80

Etbnopbarmacognosy or Sundanesse Anti-Inflammatory Plants (Moelyono MW., Supriyatna S, Anas Subamas, and Broto S.Kardono) ............................. 83

Antidiabetic Activity Of Ethanol Extract Combination From Cinnamomum burmannii Nees&Th. Nees., Trigonellafoenum-graecum L, And Morinda citrifolia L. On Alloxan Diabetic Rats (Muhtadi A, Hendriani Rand Malahayati) .......................................................................... 88

Antioxidant Activity Of Curcuma Zedoaria And Determination Oflts Curcuminoids Content (Kiki Mulkiya Yuliawati, Irda Fidrianny, Komar Ruslan Wirasutisna) ............................... 92

Cytotoxicity Of Labisia Pumila (Myrsinaceae) Extracts To Selected Human Cancer Cell Lines In Vitro (Fezah Othman and Azimahtol H.L Pihie) ...... ... ......... ... ..... .... ............. ........ ........ ........ ........ 96

Identification Of Essential Oil From Ethanolic Extract Of Cosmos caudatus Kunth. Using Gas Chromatography Massa Spectrum (GCMS) (Eka Prasasti Nur Rachmani, Soenarto, HanifNasiatul Baroroh) ....................................... 100

The Determination Of Asiaticoside Content And Screenin~ Of Acetylcholinesterase Inhibitory Potency Of Gotucola (Centella Asiatica) Harvested From Different Location (Min Rahminiwati, Latifah K. Darusman, Siti Sa'diah, Tri Wulandari) .............................. 105

Antibacterial Activity Of Beluntas (Pluchea indica L.) Extract To Vlbrio cholerae And Shigella dysenteriae Bacteria (Dewi Rusmiati, Sulistiyaningsih, Ade Zuhrotun, Widi Andayani) .................................... 111

Induction Of Callus Formation In Artemisia annua L. By In Vitro With Various Concentration Of Bap And 2,4-D (Samanhudi) ......................................................................................................................... 116

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Identification Of Ethyl Acetate Fraction Compound Of Ethanol Extract Avocado Leaves :'ersea americana Mill) 3ina L. Sari, Rini Madyastuti P, Samsul Ashari) ............................................................... 127

Development Of Phytopharmaca Product Content Of Combination Of Extract Celery (Apium Graveolens L ) And Sambiloto (Andrographis Paniculata Ness) \s Medicine For Nephrolitiasis Boesro Soebagio, Sohadi Warya, Taofik Rosdiana, Ade Zuhrotun) .................................. 132

\'alidation Of Simvastatin Analysis Method In Human Blood Plasma (In Vitro) By High Performance Liquid Chromatography With Uv Detector 1yan Sopyan, Cynthia Jaya, Driyanti Rahayu) .................................................................... 141

Antibacterial Activity Of Ethanol Extract Of Seawed Padina Australis Against \~rcobacterium Tuberculosis Strain H37rv And Resistant Streptomycin­Riphampycin Sulistiyaningsih, Tina Rostinawati, Norma Juwita) ............................................................ 147

The anti-hypertensive· effect of isolate from active fraction of roselle (hibiscus sabdariffa L) On white male rats . Yasmiwar Susilawati, Sri Adi Sumiwi, Eneng Nuraisah) .................................................. 152

Antimicrobial Activity Of Jatropha Curcas Seedcake From Biofuel Production ,Retno Wahyuningrum, Komar Ruslan Wirasutisna, Elfahmi, Marlia Singgih Wibowo) ... 159

Antibacterial Activity Of Ethanol Extract Of Noni Fruit (Morinda citri/olia L.) Against Propionibacterium acne And Multiresistant Staphylococcus aureus I Ratna Yuliani, Maryati, dan Lutta Nihawati) ..................................................................... 162

Determination Of Standard Parameter Of Ethanol Extract Of Rosemary Herb iAde Zuhrotun, Yasmiwar Susilawati, Rani Anggraeni) .................................................... 166

Analysis Of Phenolic Acids From Ketapang Bark (Terminalia catappa L.) lAde Zuhrotun l

, Asep Gana Suganda2, As'ari Nawawi2

, Komar Rustan Wirasutisna2) 171

Effects of Etbanolic Extracts from Meniran Herbs (Phyllanthus niruri L.) , Papaya leaves (Carica papaya L.), and Red Guava Leaves (Psidium guajava L.) against the Numbers of Platelets, Erythrocytes, and Hematocrytes Level on Female White Rats (Rattus novergicus) by Using Heparin Induction Method. (Ami Tjitraresmi, Eli Halimah, Yasmiwar Susilawati, Dhani Adriati Koswara) ................. 175

Proceeding ISEJ20 10

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International Seminar and Expo on Jamn ISEJ 20 I ()

Bandung. Indonesia 3-6 November 2010

IN VITRO DETERMINATION OF ANTIOXIDANT ACTIVITY OF EXTRACTS OF MAHKOTA DEWA, TEMU PUTIH, SAMBILOTO AND

KELADI TIKUS

Dyah Iswantini,I,2 Gustini Syahbirin I and Dedy Irawan I

I Department ofChemistry, Faculty ofMathematics and Natural Sciences, Bogor Agricultural University, Gedung Fapet lantai -/, JI Agathis, If'B Darmaga, Bogor. Indonesia, E-mail:

dmhpraJo(a'.l 'ahoo. co. iJ 2Biopharmaca Research Center, Bogor Agricultural University, Bogor. JI. Taman Kencana no 3

Bogar, Indonesia.

Abstract

'vfahkota dewa (Phaleria macrocarpa, Boer!), lemu pUlih (Curcuma zeodaria), sambilolo (Andrographl:~ panteu/ala, Nees) , and Ice/adi lilcus (Typhonium/lagelliji)rme) are medicinal plants that have potency as antioxidant and anticancer. The objective of this study was to determine the antioxidant capability of extract of mahkola dewa. temu pulih, sambilolo, and ke/adi tikus using tiobarbituric acid method. In this method, linoleic acid was oxidized by oxygen on 40" C for 8 days and produced malondialdehyde. The corresponding malondialdehyde have reacted with tiobarbituric acid and produced red product, the absorbance was measured on wavelength of 532 nm. The antioxidant potency of all plant extracts was monitored through its capability on inhibiting oxidation. Inhibition capability of the respective extracts on concentration of 200 ppm for aquademineralized, hot water, and ethanol extract were 83.44, 70.86, and 81.84% (mahlcota dewa), 60.21, 82.74, and 69.28% (Iemu pUlih), 81.45, 81.45, and 67.96% (sambi/olo). 68.60, 63.53, and 72.17% (ke/adi tikus), plus 87.0 I % (vitamin E), respectively. Based on analysis of variance and Duncan's test, antioxidant potency of all of extracts at concentration of200 ppm, if compared with vitamin E, were significantly different on confidence level of95%.

Keywords: Water and ethanol extracts, Mahkola dewa (Phaleria macrocarpa. Boer/), lemu pUlih (Curcuma zeodaria). sambi/ola (Andrographis panicuiala. Nees). and keladi likus (Typhonium jlage1!(/imne) , antioxidant.

INTRODUCTION avoiding carcinogens or altering their metabolism, pursuing a lifestyle or diet that

Cancer is a class of diseases in which a cell. or modifies cancer-causmg factors and/or a group of cells display uncontrolled growth, medical intervention (chemoprevention, illvasion (intrusion on and destruction of treatment of pre-malignant lesions). adjacent tissues), and sometimes metastasis Antioxidants are widely used as ingredients in (spread to other locations in the body via dietary supplements in the hope of maintaining lymph or blood). Cancer prevention is defined health and preventing diseases such as cancer, as active measures to decrease the incidence of coronary heart disease and even altitude -.:ancer. Greater than 30% of cancer is sickness. Although initial studies suggested preventable via avoiding risk factors that antioxidant supplements might promote including: tobacco, overweight or obesity, low health, later large clinical trials did not detect fruit and vegetable intake, physical inactivity, any benefit and suggested instead that excess alcohol, sexually transmitted infection, air supplementation may be harmful. In add ition pollution. This can be accomplished by to these uses of natural antioxidants in

medicine, these compounds have many

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industrial uses, such as preservatives in food and cosmetics and preventing the degradation of rubber and gasoline. Natural antioxidant agent can be found from plants such as: mahlwta dewa (Phaleria macrocarpa, Boerl), temu putih (Curcuma zeodaria), sambi/oto (Andrographis paniculata Nees), and keladi tikw; (Typhonium flagelliforme) (W ijayakusuma 1 994). Those predictions are based on the active compounds contained in those plants. According to various researches recently, mahlwta dewa flesh contains flavonoids, alkaloids, terpenoids, saponinns, and resins (Harmanto 2002; Winarto 2003).

The utilization of above plants mostly is empirical evidence based on users' experiences and the scientific evidence. Thus, the scientific research especially for the antioxidant potency of these plants is important to perform.

The purpose of the research is to determine the antioxidant activity of mahkota dewa, temu pUlih, sambi/oto, and keladi tikus in vitro by thiobarbituric acid (TBA) method. This study also is expected to give scientific informations about the antioxidant potent of these plants.

METHOD Analysis of hydroperoxide from linoleic acid by thiocyanate method (Chen el al. 1996)

Before the measurement of the antioxidant potency from each extract, it was measured the hydroperoxide as primary product of oxidized linoleic acid by thiocyanate method. This method measures the peroxide through color complex of Fe[Fe(SCN)6J. Amount of 2 mL of phosphate buffer 0.1 M pH 7.2 mL of linoleic acid 50 mM in ethanol 99.8% and I mL of deionized water were placed into dark bottle, the the mixture was incubated at 40°C. The period of incubation time was until reached the maximum absorbance. .~fter the maximum absorbance was reached, into 50 JlL of incubated mixture, it was added 6 mL of ethanol 75%, 50 JJ.L of ammonium thiocyanate 30%, and 50 JlL of FeCh 0.02 M in HCI 3.5%. After left for 3 minutes, the 3bsorbance was measure at a wavelength 482 ;lm. The measurement of hydroperoxide was -.:arried out every day until the maximum

absorbance was reached.

Analysis of antioxidant potency by TBA method (Kiuzaki & Nakatani 1993)

Analysis of antioxidant capability from each extract was carried out using serial concentration of sample test solution, namely 50, 200, and 1000 ppm. Standard solution of I, \ ,3,3-tetramethoxy propane (TMP) 6 M was diluted into serial concentration of 0.15, 0.3, 0.6, 0.75, 1.5, and 3.0 JlM. Each solution was taken I mL and added 2 mL TCA 20% and 2 mL TBA 1% (b/v) in acetate solvent 50% (v/v). Next, all of test tubes were incubated at 100°C for 10 minutes and cooled at room temperature. Once cool, the mixtures were centrifuged at speed 300 rpm then the absorbance was measure uSing spectrophotometer at a wavelength 532 nm (Yagi 1968).

Sample solution was prepared from mixture containing of 2mL of phosphate buffer pH 7.2 mL of linoleic acid 50 mM in ethanol 99.8% containing tocoferol (Vitamin E 200 ppm), and I mL of deionized water.

All of the reaction mixtures were incubated in water bath 40°C with the period of incubation time was based on the measurement of hydroperoxide from linoleic acid. Reaction mixtures were assayed for antioxidant potency after 1 or several days from the maximum absorbance of linoleic acid. Each reaction mixture was taken I mL then added 2 mL of TCA 20%, and 2 mL of TBA I % solution in acetic acid 50% solvent. Next those reaction mixtures were placed in waterbath 100°C for 10 minutes. After cool, centrifuged at 300 rpm for 15 minutes, then the absorbance was measured using spectrophotometer at a wavelength of 532 nm.

RESULTS AND DISCUSSION Linoleic acid oxidation

Antioxidant activity assay by TBA method is based on measurement of malondialdehida (MDA), level which is the end product of lipid peroxide reaction. MDA is a reactive three­carbon dialdehyde compounds. MDA generated can be measured by TBA test. MDA can react with TBA to form a product for which fluorescence and can be measured at a

41

wavelength of 532 nm. As a standard, it is used 1,1,3,3 tetrametoksipropana (TMP) that can measure MDA formed level. TMP is an MDA derivative compounds which is quite stable. Measurement of antioxidant activity with this TBA method were carried out after the level of linoleic acid oxidation was maximum because at that time also established the maximum MDA generated from lipid oxidation reactions.

Antioxidant activity of mahkota dewa, temu putih, sambiloto, and keladi tikus can be determined from its ability to inhibit oxidation of linoleic acid. The presence of secondary metabolites such as alkaloids, ftavonoids, tannins, and saponins suspected to inhibit the oxidation of linoleic acid. Hydroperoxide measurement result which was the result of oxidation of linoleic acid showed peak absorbance at 6th day.

Oxidized Linoleic acid by oxygen in the early stages will form the hydroperoxide. This hydroperoxide levels increased and after reaching the maximum level, hydroperoxides will decompose to form malondialdehyde which is the end product of lipid peroxide reaction. Malondialdehyde formation was occurred on 7th day.

Antioxidant activity analysis

Measurement of antioxidant activity was carried out on the 8th day in the hope that all hydroperoxide formed from the oxidation of linoleic acid had undergone decomposition into maIondialdehyde (MDA). Antioxidant activity of all types of plants can be seen by comparing the absorbance value that describes the concentration of MDA. Absorbance value

is proportional to the concentration of MDA and inversely proportional to the antioxidant potency. Low absorbance values indicate that plants have a high antioxidant potential. This indicates that the sample may inhibit the oxidation process (means to reduce the amount of MDA which reacts with the thiobarbituric acid to form red product).

This study used vitamin E as standard with a concentration of 200 ppm. The reason for the selection of vitamin E as a standard was at a concentration of 200 ppm inhibition percentage was near 100%. The results of Satna (2005), inhibition of vitamin E (200 ppm) by TBA method amounted to 93.0%, while the results of research Indariani (2005) the antioxidant potency of vitamin E (200 ppm) in inhibiting the oxidation of lipids up to 92.11 %. In this study, vitamin E with a concentration of 200 ppm had inhibitory amounted to 87.0 I %. The value of the inhibition was greater than aquademineral, hot water, and ethanol extract. This is because the content of antioxidant compounds in the vitamin E is more pure. From the results of standard measurements of 1,133- tetrametoxy propane (TMP), obtained the linear regression y = 0.0240 X + 0.0269, R = 99.82%. The regression equation used to calculate the MDA level of TBA test results for each extract.

Based on Salim (2006) research, obtained three extracts of mahkota dewa, that were extracted by solvent aquademineraL hot water, and ethanol. After that, the three extracts were tested antioxidant activity. Inhibition of plant extracts tested, using a solvent aquademineral, hot water and ethanol at concentrations of 50, 200, and 1000 ppm can be seen in Table 1.

Table 1 Inhibition percentage obtained from plants extracts with various solvent Extract (%)

Plant Aquademineral (ppm) Hot Water (ppm:.::2)~__E-,t.:.;..ha-,n-,-o-:-I-,-,(p,--,-p_m_-"-)-:--_ 50 200 1000 50 200 1000 50 200 1000

Mahkota Dewa 69.70 83.44 84.34 61.09 70.80 76.24 69.28 81.73 86.05 Temu Putih 44.75 60.21 72.24 71.74 82.74 86.27 54.33 69.28 79.44 Sambiloto 74.56 81.45 86.17 69.46 81.45 76.91 54.50 67.96 74.05

KeladiTikus 64.05 68.60 78.66 24.32 63.53 78.66 47.72 72.17 75.99

Antioxidant activity ofmahkota dewa extract. inhibiting the formation of MDA. The

The inhibition of mahkota dewa at a concentration of 50 ppm for all three types of

concentration of MDA in mahkota extract can be seen in Figure I.

dewa

solvent was quite large (more than 50%) in

42

.,

11 ",. ;ro"'" m ".. 9twoo! lr'ffa Extract of mahkota dewa

Figure Antioxidant activity of mahlwla dewa flesh extract.

Almost all of secondary metabolites are polar compounds. Aquademineral have a level higher polarity than ethanol. Aquademineral extract (50 and 200 ppm) of mahkotu dewa had a higher antioxidant activity than the hot water and ethanol extract. This is due to the ability of solvent extraction is directly proportional to the level of purity of the solvents. In this case aquademineral is more pure than hot water. While the ethanol extract ( 1000 ppm) had higher potency compared with aquademineral extract and hot water. This is due to the concentration of extract concentration had reached maximum standard limit (Vitamin E 200 ppm), so it is believed that there were other compounds been extracted.

\quademinerat extracts of mahkota dewu had .1 good antioxidant activity at concentration of :00 ppm because it was quite close to the \ aJue of the jnhibition of vitamin E as a positive control. Based on research Salim I :::006), the phytochemical test: alkaloids, t1avonoids, tannins, and saponins showed :~\)sitive test results. These compounds act as a ,ubstance that can inhibit lipid oxidation ~t:actjons. In Figure ), shows that the negative ~ontrol/treatment which not given the extract \\ ithout antioxidants) has a high concentration )f MDA, namely 27.2958 j.l.M. This was due to :ly,ence of compounds that could inhibit the ,;;idation process.

4ntioxidant activity of temu putih extract. \ntioxidant activity of temu putih extract also '~lOwed the value of the inhibition is quite high ; igure 2). Hot water has a higher level of "Iarity of the ethanol so that there are

.1· TC"T""TI"\.f\1 .f\

allegedly many secondary metabolites are extracted. Compounds that have a high level of polarity will be more distributed in hot water. In addition, this may also be caused by the influence of temperature and chemical compounds content ill each extract tested were different. These factors cause the type of solvent that is able to extract the maximum yield is not the same for cach type of plant extracts tested, so the results of lemu putih and mahkola dewa inhibition was different. Based on Pratiwi (2006) research, the phytochemical test: alkaloids and t1avonoids showed positive test results. These compounds are believed to act as antioxidants to inhibit lipid peroxide reaction.

:J~. "

0',".'

Extract of !'emu rutin

Figure 2 Antioxidant activity of lemu {Julih extract.

Antioxidant activity of sambi/olo extmcl. aquademineral extract of samhi/o(o (concentration 50, 200 and 1000 ppm) had the greatest inhibition when compared with hot water ethanol and extract (Figure 3), The content of secondary metabolites such as alkaloids, tannins, saponins, and flavonoids contained in the aquademineral extract was quite large. Based on Puspitasari (2006) research, the phytochemical test: alkaloids and terpenoids showed positive results for all three types of solvent. While t1avonoids showed negative results only on ethanol and tannin only in the solvent of aquademineral. Inhibition at a concentration of 200 ppm for aquademineral and hot water extract had the same value. This showed that aquademineral and hot water extract had the same inhibitory ability despite the purity level of solvent was different.

43

0"

"',...

Extract of sambi/olo

Figure 3 Antioxidant activity ofsamhilolo extract.

Antioxidant activity of each of these extracts at a concentration of 200 ppm when compared with vitamin E (200 ppm) still looked much lower (Figure 1). Despite this difference between the antioxidant potency of each extract at a concentration Of 50, 200, and 1000 ppm was not so visible. This was because the concentration of the extract was too large and not proportional to the concentration of linoleic acid substrate.

Antioxidant activity of keladi tiku.~ extract. Aquademineral extract of keladi tikus (50 ppm) had higher antioxidant activity than the hot water and ethanol extract (Figure 4). This is because aquademineral has a high level of polarity than ethanol and more pure than hot water, so that alleged some secondary metabolites such as alkaloids and flavonoids that have a high level of polarity are also distributed in it. Based on Affandi (2006) research, the phytochemical test: alkaloids, tlavonoids, and tannins showed positive results. These compounds are an antidote compound which can inhibit free radical oxidation reactions.

Dc, «: 0'" '".',' ~ 11l .,"I'0 c 1)1Qe I c III S

~ C o o

lJil'l'"' 1MlIl'l""

Extract of !tel"". 1'*0$

Figure 4 exlract.

Antioxidant activity of keladi tikus

At a concentration of 200 ppm ethanol extraction had a higher inhibitory This was due to the compounds contained in extracts of keladi tikus distributed more on ethanol, although the level of polarity lower than aquademineral and hot water. Based on these results we can conclude that the inhibition of antioxidants is not always proportional to the level of purity and polarity of the solvent used.

Meanwhile, at a concentration of 1000 ppm. the inhibition of aquademineral and hot water had the same value but higher than ethanol. This was due at concentrations above the maximum limit (200 ppm) the extract had different antioxidant potential which was less visible, making it difficult to distinguish the influence of the purity and polarity of a solvent.

Based on the results obtained, it was concluded that mahkoJa dewa, lemu pulih, samhiloto, and keladi tikus with three kinds of extracts, namely aquademineral. hot water and ethanol were very potent antioxidants. This can be seen at a concentration of 200 ppm, the value of the inhibition of each extract was close to the value of the inhibition of vitamin E.

Alkaloids and flavonoids gave positive results on all types of plants tested (Salim 2006). Flavonoids and alkaloids is a good reductor compound. Flavonoids act as a good container for hydroxyl and superoxide free radicals (Robinson 1995). Accord ing to Mangan (2003), flavonoid substance served as an antidote to free radicals that could disnJpt the body's balance system and could lead to cancer.

Tannins are compounds that contained in the tea plant. Based on the Yen (1995) research, it reported that various types of tea had antioxidant activitv. Based on the results of the study also tannin~ could inhibit the process of mu~tion ~nd cancer, and free radicals and induced enzymes that act as antioxidants.

Saponins in the plant have been known to be used for treatment. Saponin contained in samples plants (Physalis angulola Linn.) had efficacy as an antitumor and inhibits the growth of cancer especially colon cancer (Mangan 2003). In addition, sap<mins

44

contained in kunyil, tapak dara, sabung nyawa, mengkudu, kitolod, and pegagan had efficacy as anticancer.

AN OVA and Duncan's Test

The statistical test used for the antioxidant potency were ANOVA and DUNCAN. The tests were only done at a concentration of 200 ppm for each extract of akuademineral, hot water, and ethanol extract, which were directly compared with vitamin E (200 ppm).

ANOV A test of mahkota dewa, temu putih, sambi/olo, and keladi tikus extract showed significantly different results. Therefore it was necessary to do further test using DUNC AN test to see whether there were differences in antioxidant potency in aquademineral, hot water, ethanol extract and vitamin E on each plant. DUNCAN test showed different results from each plant extract. This was because the content of secondary metabolites contained in extracts were difTerent that the inhibition power against MDA tonnation was also different.

CONCLUSION

The extract of mahkota dewa, lemu pUlih, sambil%, and keJadi likw; had potency as antioxidants based on their ability to inhibit the fonnation of MDA. Data results showed that the smallest inhibition was found in keladi tikus extract of 24.32% at concentrations of 50 ppm with hot water solvent. While the greatest inhibition found in /emu pulih extracts at 1000 ppm, namely 86.27% with the same solvent.

ANOVA of extract at a concentration of 200 ppm for aquademineral, hot water, ethanol extract, and vitamin E showed significantly different results. DUNCAN statistical test showed that the antioxidant activity was different.

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Chen J e/ al. 1996. The Protective effect of tea on cancer: Human Evidance. Di dalam Phy/ochemicals As Bioactive Agents. Basel: Technomic Publishing Co., inc.

Hannanto N. 2002. Mahkota Dewa Obat Pusaka Para Dewa. Jakarta: Agromed ia Pustaka.

Indariani S. 2005. Uji aktivitas antioksidan ekstrak daun jambu biji (Psidium guajava L.). Pusat studi Biotar­maka,LPPM-IPB.

Kikuzaki H & Nakatani N. 1993. Antioxidant effect of some ginger constituents. Journal ofFood Science 58(6): 1407­14]0.

Mangan. 2003. Cara bijak Menaklukan Kanker. Jakarta:Agromedia Pustaka.

Pratiwi W. 2006 . Penetuan daya inhibisi ekstrak air dan etanol temu putih (Curcuma zeodaria) terhadap aktivitas tirosin kinase secara in vitro. [Skripsi]. Bogor: Fakultas Matematika dan Jlmu Pengetahuan Alam, Institut Pertanian Bogor.

Puspitasari MS. 2006. Inhibisi ekstrak air dan etanol sambiloto (Andrographis paniculala) terhadap aktivitas tirosin kinase secara in vi/ro. [Skripsi]. Bogor: Fakultas Matematika dan IImu Pengetahuan Alam, Inslitut Pertanian Bogor.

Robinson T. 1995 . Kandungan Organik Tumbuhan Tinggi. Bandung: Penerbit ITB.

Salim. 2006. Penentuan daya inhibisi ekstrak air dan etanol daging buah mahkota dewa (Phaleria macrocarpa (Scheff.)Boer1.) terhadap aktivitas tirosin kinase secara in vitro. [Skripsi]. Bogor: Fakultas Matematika dan IImu Pengetahuan Aiam, Institut Pertanian Bogor.

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