ACTIVE COMPOUND CONTENT OF SOUR STAR FRUIT LEAVES METHANOL EXTRACT

ACTIVE COMPOUND CONTENT OF SOUR STAR FRUIT LEAVES METHANOL EXTRACT

(Averrhoa carambola L.) IN BALINESE TRADITIONAL FOOD

Ni Putu Adriani Astiti, Sang Ketut Sudirga and Yan Ramona

Biology department

Faculty of Mathematics and Natural Sciences Udayana University, Bali

2019

INTRODUCTION

Degenerative disease

cancer

coronary heart attack

Deabetes melitus

Free Radical And

oxidative stress

spices, tea, chocolate, seeds of plants, vegetables, enzyme, and other proteins

are sources of natural antioxidant compounds

Natural antioxidant sources are

dominated by plants that contain

flavonoid and phenolic compounds

spreading in all parts of the plants.

The leaves of star fruit plants (Averrhoa carambola) have been

traditionally used to cure many diseases, such as hypertension and fever.

the leaves of this plant have also widely used as the main raw material in lawar making.

Degenerative disease

Leaves of sour and sweet fruit star plants are used as raw

materials lawar making in Bali and often served in any Balinese

traditional events, such as wedding ceremony.

In this research, the methanol extract of this plant leaves was analyzed and identified for active compound

contents

The main objective of this research was to investigate

types of active compounds contained in such leaves

previously extracted in methanol.

MATERIALS AND METHOD Extraction

These materials (Leaves of sour fruit star plants ) were air dried, powdered with a warring blender, and macerated with

methanol solution

The filtrate was then evaporated in a vacuum rotary evaporator at 50oC to concentrate the extract and purified in a chromatography

column using eluent with various polarities ^. The choice of eluent was based on like

dissolves like. The moving phase eluents used were100% n-hexane, n-hexane :

dichloromethane (1:1), 100%

dichloromethane, dichloromethane : ethyl acetate (1:1), 100% ethyl acetate, ethyl acetate : ethanol; (1:1), 100% ethanol,

The active

components of extract

of sour star fruit plant

was analysed using a

GC MS instrumentation

RESULTS AND DISCUSSION

The active compounds contained in the methanol extract of sour star fruit leaves are shown

Butane, 1,1-diethoxy-3methyl-(CAS)1.1-DH, Decanoic acid ethyl ester (CAS) Ethyl decano, Hexadecanoic acid, ethyl ester (CAS) Ethyl,

Eicosanoic acid methyl ester (CAS) Arachidic, and

9- Octadecanoic acid methyl ester (E)-(CAS)

Butane,1,1-diethoxy-3-methyl,

this compound can be used to synthesize

organics, production of ethylene, fuel, mix of petroleum, and raw material for ethylene and butadiene as well as main raw material of

synthetic rubber .

Dodecanoic acid, methyl ester

This compound is used as an alternative food flavour enhancer (Alibaba, 2019).

Hexadecanoic acid, ethyl ester

This compound has other name of stearate acid which is commonly used as food

supplement, cosmetics, and industrial products. this compound is used as a

material in wax, plastic, and soap making in

addition to soften rubber.

9-Octadecenoic acid, methyl ester

Other name of this compound is oleic acid. This is the most

important saturated fatty acid in the production of biodiesel.

Biodiesel with high saturated bond has high octan number and resistant to antioxidant. This compound can be used to

produce good quality biodiesel ( Gultom, 2011). Besides, this

compound is also used as bio- pesticide.

,

Eicosanoic acid methyl ester (CAS) Arachidic, This compound is also known as arakida acid.

This acid is used in the detergent production, as

photographic material, and lubricant.

CONCLUSION

Ten compounds were identified in the methanol extract of sour star fruit leaves using GC-MS

instrumentation. these include:

Butane, 1,1-diethoxy-3methyl-(CAS)1.1-DH, Decanoic acid ethyl ester (CAS) Ethyl decano, Hexadecanoic acid, ethyl ester (CAS) Ethyl,

Eicosanoic acid methyl ester (CAS) Arachidic, and

9- Octadecanoic acid methyl ester (E)-(CAS)

THANK YO ^U

1

ACTIVE COMPOUND CONTENT OF SOUR STAR FRUIT LEAVES METHANOL

EXTRACT (Averrhoa carambola L.) IN BALINESE TRADITIONAL FOOD

Ni Putu Adriani Astiti¹⁾, Sang Ketut Sudirga²⁾ and Yan Ramona³⁾

(1,2)Biology department, Faculty of Mathematics and Natural Sciences, Udayana University, Bali

(3)Integrated Laboratory for Biosciences and Biotechnology, Udayana University Corresponding author E-mail: adriani@unud.ac.id

Abstract

Lawar is a Balinese traditional food that can be prepared from various materials, including leaves of sour star fruit plants. This type of food commonly served during religious related events particularly in south parts of Bali island, Badung regency and Denpasar city. Antioxidants are compounds with ability to scavenge free radicals in human body so that they can be used as an alternative method to avoid people from having various diseases due to free radicals. Therefore, the content of active compounds in such plant leaves needs to be investigated so that their use to cure various degenerative diseases can be well understood.In our current research, leaves active compound content of such plant was screened and identified by applying Prior to screening and identification, the leaves were extracted by applying maceration method followed by purification by a pplying thin layer chromatography and column chromatography. The results showed that 5 active compounds including Butane, 1,1- diethoxy-3methyl-(CAS)1.1-DH, Decanoic acid ethyl ester (CAS) Ethyl decano, Hexadecanoic acid, ethyl ester (CAS) Ethyl pa,Eicosanoic acid methyl ester (CAS) Arachidic, and 9- Octadecanoic acid methyl ester (E)- (CAS) were found in the methanol extract of the plant leaves. In the phytochemical screening flavonoid and tannin content of such extract was relatively high and is potential as antioxidant to scavenge free radicals.

Keywords: sour star fruit, methanol extract, Lawar, Balinese food

INTRODUCTION

Leaves of sour star fruit plant (Averrhoa carambola L.) have been used as the main raw material in making lawar (a Balinese traditional food) commonly served in various religious related events. Such traditional food is one of the most favorite foods among Balinese people as it has specific delicious taste. This food is also known to contain high antioxidant compounds (.2).

Oxidative stress induced by free radicals can cause various degenerative diseases, such as cancer, coronary heart attack, and early aging process. Human body needs sufficient amount of defence system to fight the free radical activities to induce such diseases. Compounds with ability to scavenge free radicals are antioxidant that can be obtained from our foods. Free radicals are molecules with unpaired electron on their outermost electron orbit. This condition makes them become very reactive molecules. Free radicals tend to undergo chain reactions in human body and result in continuous damages in long term (5). Human body has endogenous defence system to fight such free radicals. This particularly happens through metabolisms of normal and inflamed cells. The level of free radicals may increase due to stress factors, radiation, smoke of cigarettes, and environmental pollutions. Such conditions will result in reduced defence system of human body, and therefore additional antioxidants are needed to fight the high level of

2 free radicals. Based on the above background, leaves of sour star fruit plant known to contain high level of antioxidant and used in lawar making in Bali need to be considered for investigation. Lawar made from such leaves is believed by Balinese people to improve their health. Pharmacologic potential of such leaves has not been well understood, and therefore identification of active compounds contained in the extract of such leaves is urgently needed.

MATERIAL AND METHODS

Air dried powder of sour star fruit leaves was macerated in methanol by applying a method specified by(6). A quantity of 100 gram leaf powder was extracted in 500 ml methanol on a shaker for 2 hours, macerated for 24 hours, filtered, and evaporated at 50 oC in a vacuum evaporated to obtain concentrated extract in the form of pasta. Purification of such concentrated extract was performed by applying column chromatography suing eluent with various polarities as mobile phase. The choice of mobile phase was based on the principle of like dissolves like. Mobile phase used in this experiment included n-heksan, n-heksan:diklorometan (1:1), diklorometan, diklorometan:etil asetat (1:1), etil asetat, etil asetat: etanol (1:1), etanol, etanol:metanol (1:1), and metanol. The purified concentrated extract of the leaves was analysed for active compound content by applying GC MS instrumentation. Besides, screening of phytochemical compounds on the concentrated extract was also conducted in order to investigate the potential of the extract.

RESULTS AND DISCUSSION

GC-MS results showed that the methanol extract of sour star fruit leaves was identified to contain 5 active compounds.

These included Butane, 1,1-diethoxy-3methyl-(CAS)1.1-DH, Decanoic acid ethyl ester (CAS) Ethyl decano, Hexadecanoic acid, ethyl ester (CAS) Ethyl pa,Eicosanoic acid methyl ester (CAS) Arachidic, and 9- Octadecanoic acid methyl ester (E)-(CAS)

an explanation of each active compound is Butana,1,1-diethoxy-3-methyl,

This compound has retention time of 3.590 minutes which is identical to mass spectra of WILEY7.LIB database.

According (19) this compound can be used for synthesis of organics, production of ethylene, fuel, and as a mixture in petroleum, as well as a main raw material in the production of synthetic rubber.

Decanoic acid ethyl ester

This compound has retention time of 10.016 minutes which is identical to mass spectra of WILEY7.LIB database.

Saturated fatty acid (miristate acid) is good for soap making. Fatty acid with 14-18 carbon atom is excellent for foam producers. If it is used in soap making, it will produce hard soap, high level of detergency and produce soft foam (3).

It is also a compound to compose essential oil.

Hexadecanoic acid, ethyl ester

This compound has several different names, such as stearic acid that is commonly used as food additive, cosmetic, and industrial products. In industrial sectors, this compound is used as material in the making candles, soaps, plastics,

3 cosmetics, as well as to soften rubbers. this compound also help to harden soaps. Excessive use of this compound in soap making can cause less foamy ( 7 ).

Eicosanoic acid methyl ester

This compound is also known as arakida acid. This acid is used in the detergent production, as photographic material, and lubricant. This compound is a saturated fatty acid with 20 carbon atoms. This is minor constituent of cupuaçu butter (7%), [2] perilla oil (0-1%), [3] peanut oil (1,1-1,7%), [4] corn oil (3%), [5] and cocoa butter (1%).

[6] This is also 7.08% of fat in species of Durio graveolent.

9- Octadecanoic acid methyl ester

The retention time of this compound is 17.924 minutes which is identical to (E)-9-Octadecanoic Acid, ethyl ester according to database of NIST08.LIB. The molecular formula of this compound is C20H38O. This compound is similar to steroid. It can be used as lubricant and plasticizer (8).

There were 6 peaks produced in the chromatogram of GC-MS, and 2 of those peaks (peak 3 and 5) were the same compound (Hexadecanoic acid, ethyl ester), but had different retention times (12.543 and 16.882 minutes). This indicated that times needed by these compounds to migrate were different in different columns. Phytochemical screening showed that the leaf extract of sour star fruit plant contains flavonoid and tannin, and indicates that it is potential as an antioxidant source to scavenge free radicals so that appearance of degenerative diseases can be prevented. Oxidative activity of flavonoid is due to its ability to donate hydrogen atom or to chelate metal ions. Some flavonoid compounds, such as quercetin, kaempferol, myricetin, apigenin, luteolin, vitexin and isovitexin are found in cereals, vegetables, fruits, and their derivate. This strengthen hypothesis that flavonoid has certain biological effects which are related to its anti-oxidative properties (17 ). A Food with flavonoid in it can be used as an alternative medicine due to its anti-carcinogenic properties. This can preve. people from having viral infection, cancer, diabetes, or depression (17 )Tannin on the other side is a complex compound with mixtures of polyphenols which is difficult to separate. Phenol is an important compound with ability to chelate metal ions (17 )The mechanism or the process of chelating depends on the pH of this phenolic compound (11). This normally happens in hydrolysed tannin and resulting in more stable chelated metal in human body. Tannin will have specific acidic astringent taste when diluted in water.

This high content of tannin in sour star fruit plant affects its sour taste in its fruits which much sour than that of sweet star fruit plant.

4 CONCLUSION

1. Methanol extract of sour star fruit leaves (Averrhoa carambola L.) is potential as antioxidant as they contain high level of flavonoid and tannin.

2. Based on the GC-MS chromatogram, 5 active compounds were identified, including Butane, 1,1-diethoxy- 3methyl-(CAS)1.1-DH, Decanoic acid ethyl ester (CAS) Ethyl decano, Hexadecanoic acid, ethyl ester (CAS) Ethyl pa, Eicosanoic acid methyl ester (CAS) Arachidic, and 9- Octadecanoic acid methyl ester (E)-(CAS)

Acknowledgment

This research was supported by the Directorate Research and Community Service (DP2M), Directorate of Higher Education through

Higher Education Primary Research (PDUPT) program 2018

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