ANALYSIS AND IDENTIFICATION OF ACTIVE COMPOUNDS IN METHANOL EXTRACT OF SWEET STAR FRUIT (Averrhoa
carambola L.) LEAVES, COMMONLY USED AS RAW MATERIAL OF LAWAR (A BALINESE TRADITIONAL FOOD )
Ni Putu Adriani Astiti 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 sweet 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 sweet star fruit
plant was analysed using a GC MS
instrumentation
RESULTS AND DISCUSSION
The active compounds contained in the methanol extract of sweet star fruit leaves are shown
Butane, 1,1-diethoxy-3-methyl Dodecanoic acid, methyl ester Dodecanoic acid ethyl ester
Pentadecanoic acid, ethyl ester Hexadecanoic acid, methyl ester Hexadecanoic acid, ethyl ester
Octadeca-9,12-dienoic acid methyl
9-Octadecenoic acid, methyl ester
Octcadecanoi acid, methyl ester
9-Octadecenoic acid ethyl ester
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).
Dodecanoic acid ethyl ester
are good for raw material to make hard soap as they will produce foamy soap with high detergency (Cavitch, 2001). This
compound is also a component of essential oil.
Pentadecanoic acid, ethyl ester
this compound is a saturated fatty acid, where most of saturated fatty acid have antifungal and anti-bacterial activity.
Hexadecanoic acid, methyl ester
belong to fatty acid group with capacity as anti-bacteria by disrupting bacterial cell wall and cell membrane. This
compound can work synergically with
various active compounds and increase its anti-bacterial activity (Padmini et al,
2010)
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.
Octadeca-9,12-dienoic acid methyl
This compound is used in various industries, such as cosmetic,
soap, shampoo, fabric softener, paint, dan plastic. In kepoh
plant, sterculic acid is also used as adaptive biodiesel (Duengo, 2011).
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-
Octadecanoic 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.
9-Octadecenoic acid ethyl ester
This is steroid-like compound which is
useful for lubricant and plasticizer
(Fessenden dan Fessenden.2000).
CONCLUSION
Ten compounds were identified in the methanol extract of sweet star fruit leaves using GC-MS instrumentation. these include:
Butane, 1,1-diethoxy-3 methyl-(CAS)1.1-DII Dodecanoic acid, methyl ester(CAS) methyl
Dodecanoic acid, methyl ester(CAS) Ethyl Laun Pentadecanoic acid ethyl ester
Hexadecanoic acid methyl ester (CAS) Methyl pa Hexadecanoic acid, ethyl ester
Octadeca 9.12 dienoic acid methyl
9- Octadecenoic acid methyl ester ( E )-(CAS) Octadecanoic acid methyl Ester
9-Octadecanoic acid ethyl ester
1
ANALYSIS AND IDENTIFICATION OF ACTIVE COMPOUNDS IN METHANOL EXTRACT OF SWEET
STAR FRUIT (Averrhoa carambola L.) LEAVES,
COMMONLY USED AS RAW MATERIAL OF LAWAR (A BALINESE TRADITIONAL FOOD)
Ni Putu Adriani Astiti1) and Yan Ramona1,2)
(1)Biology department, Faculty of Mathematics and Natural Sciences, Udayana University, Bali
(2)Integrated Laboratory for Biosciences and Biotechnology, Udayana University
*Corresponding author E-mail: [email protected]
ABSTRACT
The leaves of star fruit plants (Averrhoa carambola) have been traditionally used to cure many diseases, such as hypertension and fever. Besides, the leaves of this plant have also widely used as the main raw material in lawar making. 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. Analysis was conducted by applying GC-MS instrumentation using methanol solution. The results showed 10 active compounds (Butane, 1,1-diethoxy-3 methyl-(CAS)1.1-DII, Dodecanoic acid, methyk ester(CAS) methyl, Dodecanoic acid, methyl ester (CAS) Ethyl Laun, Pentadecanoic acid ethyl ester, Hexadecanoic acid methyl ester (CAS) Methyl pa, OCTADECA 9.12 DIENOIC ACID METHYL, 9- Octadecenoic acid methyl ester ( E )-(CAS), Octadecanoic acid methyl Ester, and ( E ) 9-Octadecanoic acid ethyl ester) identified in methanol extract of such leaves.
Key word: Averrhoa carambola, active compounds, lawar, Balinese traditional food.
INTRODUCTION
Sweet star fruits plants are fruiting all seasons for the whole year. The fruits of this plants contain high level of vitamin A and C and have potential as antioxidant properties. Leaves of this plant have been used emphirically to cure stomachace, parotitis, as well as rheumatic related diseases (Junita, 2010). In Bali such leaves have also been used as raw material for lawar (a traditional Balinese food) making. Such traditional food is commonly sereved in any Balinese ceremonial evets, such as temple festival or weding ceremony.
Herbal medicines have been developed worldwide by exploiting potential plant components. This indicates that people tend to be back to nature to achieve optimal healthiness to cope with various diseases naturally. Radical-induced oxidative stress has been recognized to stimulate various degenerative related diseases, such as cancer, coronary heart attack or early aging. Human body frequently need additional antioxidant to fulfil the body need, and this can often be obtained from plant materials. Junita (2010) reported that there is an inversed relationship between dose of leaf extract of sweet star fruit pland and sugar blood level of rats.
Free radicals are molecules with unpaired electron on their outermost orbit that make them very reactive. Such free radicals tend to initiate chain reactions within human body and cause damages continuously.
Under normal conditions, human body has endogenic defence system against free radicals. This is normally done by cellular metabolism of normal cells and inflammation. Stress factors, radiation, smoke of cigarette, and pollution can increase the quantity of free radicals in our body. If this happens, our body need extra antioxidant from our diets to neutralize those excessive free radicals. As a mega biodiversity country, Indonesia has various types of plant with high antioxidant content. Star fruit plants are examples of Indonesian flora that need to be explored as antioxidant sources. The leaves of those plants are believed to improve human health by Balinese people. The knowledge pharmacological potency of such leaves is still limited. Therefore identification of active compounds content of sweet star fruit leaves need to be conducted. The main objective of this research was to investigate
2
types of active compounds contained in the extract of leaves of such plants, so that their potency for health improvement can be recognized.
METHODS
This is an explorative research using methanol extract of sweet star fruit leaf powder. Extraction was conducted by maceration method specified by Fauziati (2017 ). A quantity of 50g leaf powder was extracted with 250 ml methanol, shaked for 2 hours, macerated for 24 hours, and filtered. 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, ethanol : methanol (1:1), and 100% methanol. The amount of each eluent used was 50 ml. The active components of extract of sweet star fruit plant was analysed using a GC MS instrumentation.
RESULTS AND DISCUSSION
The active compounds contained in the methanol extract of sweet star fruit leaves are shown in Table 1. Ten compounds appeared following GC-MS analysis.
Table 1: Compounds appeared following GC-MS Analysis
No. Name Retention time
(minute)
Area ( % )
Height of peaks
1 Butane, 1,1-diethoxy-3-methyl 3.594 8326807 4264212
2 Dodecanoic acid, methyl ester 11.697 844179 525359
3 Dodecanoic acid ethyl ester 12.551 1202825 831235
4 Pentadecanoic acid, ethyl ester 14.824 576038 395694
5 Hexadecanoic acid, methyl ester 16.215 6287117 4587809
6 Hexadecanoic acid, ethyl ester 16.890 2027551 1400658
7 Octadeca-9,12-dienoic acid methyl 17.868 353790 232168
8 9-Octadecenoic acid, methyl ester 17.933 2565251 1749246
9 10
Octcadecanoi acid, methyl ester 9-Octadecenoic acid ethyl ester
18.165 18.545
538420 622177
352787 416128
Butane,1,1-diethoxy-3-methyl,
This compound has retention time of 3.594 minutes which is identical with mass spectra of WILEY7.LIB database. According to Jong ( 2015 ), 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 has retention time of 11.697 minutes identical to mass spectra of WILEY7.LIB database. This compound is used as an alternative food flavour enhancer (Alibaba, 2019).
Dodecanoic acid ethyl ester
This compound has retention time of 12.55 minutes identical to mass spectra of WILEY7.LIB database. Saturated fatty acid (miristate acid) is for soap making. For Fatty acids with 14-18 carbon atoms are good for raw material to make hard soap as they will produce foamy soap with high detergency (Cavitch, 2001). This compound is also a component of essential oil.
Pentadecanoic acid, ethyl ester
This compound has retention time of 14.824 minutes identical to mass spectra of WILEY7.LIB database. According to Agoramoorthy et al (2007), this compound is a saturated fatty acid, where most of saturated fatty acid have antifungal and anti- bacterialactivity.
3 Hexadecanoic acid, methyl ester
This compound has retention time of 16,890 minutes identical to mass spectra of WILEY7.LIB database. Hexadecanoic acid methyl ester (metil palmitate) belong to fatty acid group with capacity as anti-bacteria by disrupting bacterial cell wall and cell membrane.
This compound can work synergically with various active compounds and increase its anti-bacterial activity (Padmini et al, 2010)
Hexadecanoic acid, ethyl ester
This compound has other name of stearate acid which is commonly used as food supplement, cosmetics, and industrial products.
In the stearic acid industry, this compound is used as a material in wax, plastic, and soap making in addition to soften rubber.
Stearic acid is also used to harden soap. Excessive use of stearic acid causing the soaps less foamy while deficient in stearic acid in soap making will cause the soap stay liquid (Hart et al, 2003).
Octadeca-9,12-dienoic acid methyl
This compound has retention time of 17.868 minutes identical to mass spectra of WILEY7.LIB database. Other name of this compound is sterculic acid. Plant-based oil consists of sterculic acid with molecular formula of C19H3402. This compound is used in various industries, such as cosmetic, soap, shampoo, fabric softener, paint, dan plastic. In kepoh plant, sterculic acid is also used as adaptive biodiesel (Duengo, 2011).
9-Octadecenoic acid, methyl ester
This compound has retention time of 17.933 minutes identical to mass spectra of WILEY7.LIB database. 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.
Octadecanoic acid, methyl ester
This compound has retention time of 18.165 minutes identical to mass spectra of NIST 08.LIB database. This compound plays important role as insecticide, effective to control larvae of Aedes aegyptii and Culex pipiens pallens (Imaniar et al., 2013;
Perumalsamy et al., 2015).
9-Octadecenoic acid ethyl ester
This compound has retention time of 18.545 minutes identical to mass spectra of NIST 08.LIB database. This is steroid-like compound which is useful for lubricant and plasticizer (Fessenden dan Fessenden.2000).
CONCLUSION
Ten compounds were identified in the methanol extract of sweet star fruit leaves using GC-MS instrumentation. these include:
Butane, 1,1-diethoxy-3 methyl-(CAS)1.1-DII Dodecanoic acid, methyk ester(CAS) methyl Dodecanoic acid, methyk ester(CAS) Ethyl Laun Pentadecanoic acid ethyl ester
Hexadecanoic acid methyl ester (CAS) Methyl pa Hexadecanoic acid, ethyl ester
Octadeca 9.12 dienoic acid methyl
9- Octadecenoic acid methyl ester ( E )-(CAS) Octadecanoic acid methyl Ester
9-Octadecanoic acid ethyl ester
All of those compounds are potential for anti-bacteria and anti-fungi. Besides, they are also used as solvents in pharmaceutical aspects, antioxidant, material for soap making, biodiesel, lubrican, anti-mosquito,and food flavour enhancer.
4 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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