Averrhoa Carambola, L. Leaf Extract: Raju V.K., 2003

Averrhoa carambola, L.
leaf extract
INTRODUCTION
Medicinal plants, the World’s oldest known healthcare products,
play a key role in traditional medicines. They are not only used for
primary health care, many widely used pharmaceuticals are derived from
plants and other natural resources (G.R.K.Sharma, 2006).
The history of medicine dates back perhaps to the origin of human
race. Rig veda written between 4500-1600 BC is the oldest repository of
human knowledge in India. Later, Suhsruta Samhita and Charaka Samhita
dealt with various drugs (Jain,S.K., 1987). The Sushruta Samhita contains
184 chapters and description of 1120 illnesses, 700 medicinal plants, a
detailed study on Anatomy, 64 preparations from mineral sources and 57
preparations based on animal sources (Raju V.K., 2003). The Charaka
Samhita was recorded several thousand years ago from the
Teachings of the sage Punarvasu Atreya. The Charaka Samhita is
believed to have arisen around 400-200 BC. Charaka contains over 8,400
metrical verses, which are often committed to memory, in toto, by
modern medical students of Ayurveda. It presents most of the theoretical
edifice of Ayurveda and concentrates on the branch of Ayurveda called
1
Averrhoa carambola, L. leaf extract
kayachikitsa (internal medicine) (Menon, I A., et al., 1969).
Traditional healthcare systems using medicinal plants can be
recognized and used as a starting point for the development of novelties
in drugs. The volatile components of medicinal plants and herbs are
extracted that can be used for various purposes.
WHO had listed over 21,000 plant species used around the World
for medicinal purposes (NBPGR Booklet, 2006). Overall 119 plant
derived prescription drugs are commonly used in different countries, 74%
of which were discovered due to chemical isolation of active compounds
of plants used in traditional system of medicine (Sharma,G.R.K., 2006).
India is ranked no.8 in World Biodiversity. About 70% of India’s
medicinal plants are found in Tropical area and the rest in temperate and
alpine areas. Macrostudies showed that a large percentage of known
medicinal plants occur in the dry and moist deciduous vegetations as
compared to the evergreen or temperate habitats (Tiwari, 2000).
In some countries like India, pharmaceutical companies are already
marketing preparations of tablets, capsules, etc. they are made directly
from appropriate plant extracts for the treatment of specific diseases like
Hepatitis. Medicinal plants and herbs contain substances known to
modern and ancient civilizations for their healing properties. Until
2
development of chemistry, particularly, of the synthesis of organic
compounds in the nineteenth century, medicinal plants and herbs were
sole source of active principles capable of curing man’s aliments. They
continue to be important to people that do not have access to modern
medicines and moreover, modern pharmaceuticals rely heavily on the
same active principles (Yusuf et al., 1999).
Drug discovery, ethnobotany and traditional and indigenous
medicines have long been basic to medicinal plant research. As new uses
for medicinal plants have been discovered and popularized, sustainability
has become increasingly an issue; concern over the growth in biopyracy
goes hand in hand with the critical need for conservation of both species
and habitat (Lewis, et al., 2003).
The present study is concentrated on Averrhoa carambola, L. of
the family Geraniaceae. This plant is most common in African countries
and its fruits are used as a vegetable and in desserts. In India, even though
it is a native species, is less common among people as a food source. But
the analysis of star fruits revealed the presence of high amount of Dietary
fibers, protein, Vitamin C, Potassium, etc.(Facciola, et al., 1990). The
phytochemical screening as well as antimicrobial and anthelmintic assays
was carried on the leaves to reveal its potential as a good source of drug.
3
Plant Description
1.1. Taxonomy
Current name: Averrhoa carambola (The generic name is after
Averrhoes (1126-98), the widely known Arab Philosopher. The specific
name, ‘carambola’, is said to have come from Malabar and was adopted
early by the Portuguese).
Authority : L.
According to Bentham and Hooker’s classification
Kingdom : Plantae
Class : Dicotyledonae
Sub-class : Polypetalae
Series : Disciflorae
Cohort : Geraniales
Family : Geraniaceae
4
K5C5A(5+50) G(5)
Synonym(s)
Averrhoa acutangula., Stokes
Sanskrit Synonyms
Karmaranga, Karuka
1.2. Common names
Malayalam : Chaturappuli, Irumbanpuli, Karappola
Hindi : kamrakh, kamranga
English : carambola, star fruit, Carambola apple, Karazola,
Coromandal gooseberry
Filipino : balimbing
French : blinblin longue, carambolier, cornichon du pays
Indonesian : belimbing manis
Malay : belimbing manis
Mandarin : yongt’o
5
Spanish : carambola, carambold, carambole, jalea
Thai : ma fueang
Vietnamese : khê
1.3. Morphology
Tree to 30 ft (Fig.1); young parts finely pubescent; leaf 5-11,
ovate-elliptic, short pointed, mostly 1-2 in. long, nearly glabrous beneath;
flowers in the leaf axils(Fig.2); petals about ¼ in. long, white-marked
purple; 5 stamens without anthers; fruit 2-5 in. long, ovoid or ellipsoid, 3-
5 deep ribs, yellow or yellow brown, very pulpy, mildly acidic or even
sweet when ripe, fragrant (Fig.3&4)(Bailey,L.H., 1948)
1.4. HABITAT
Grows best in the hot, humid tropics but will tolerate some cool
weather. Young plants may be killed or badly damaged by frost, while
mature specimens can withstand temperatures as low as -30 C for short
periods, with some damage to branches and leaves. They can also tolerate
dry periods and some wind if it is not too cold.
1.5. Geographic distribution
Native : Indonesia, Malaysia, India, Sreelanka
Exotic : Australia, Brazil, Cambodia, China, Colombia, Dominican
Republic, Israel, Japan, Laos, Myanmar, Philippines,
6
Singapore, Taiwan, Province of China, Thailand, Trinidad
and Tobago, Uganda, United States of America, Vietnam
1.6. Biophysical limits
Altitude : 0-900 m
Soil type : Prefers deep, well-drained clay loams but can grow
successfully on sandy soils and heavy clays.
Ayurvedic Medicinal Properties
Rasa : Amla
Guna : Guru, Snigda
Virya : Ushna
1.7. Medicinal Properties of the Plant as per Ayurveda
Plant pacifies vitiated kapha, pitta, skin diseases, poison, pruritus,
worm infestations, diarrhea, vomiting, hemorrhoids, intermittent fever,
over perspiration and general debility.
In India, the ripe fruit is administered to halt hemorrhages and to
relieve bleeding hemorrhoids. The dried fruit or the juice may be taken to
counteract fevers.
Other medicinal uses
A conserve of the fruit is said to allay biliousness and diarrhea and
7
to relieve a"hangover" from excessive indulgence in alcohol. A salve
made of the fruit is employed to relieve eye afflictions. In Brazil, the
carambola is recommended as a diuretic in kidney and bladder
complaints, and is believed to have a beneficial effect in the treatment of
eczema. In Chinese Materia Medica it is stated: "Its action is to quench
thirst, to increase the salivary secretion, and hence to allay fever." A
decoction of combined fruit and leaves is drunk to overcome vomiting.
Leaves are bound on the temples to soothe headache. Crushed leaves and
shoots are poulticed on the eruptions of chicken-pox, also on ringworm.
The flowers are given as a vermifuge. In southeast Asia, the
flowers are rubbed on the dermatitis caused by lacquer derived from Rhus
verniciflua, Stokes. Burkill says that a preparation of the inner bark, with
sandalwood and Alyxia sp., is applied on prickly heat. The roots, with
sugar, are considered an antidote for poison. Hydrocyanic acid has been
detected in the leaves, stems and roots. The powdered seeds serve as a
sedative in cases of asthma and colic. (Morton,J., 1987).
8
REVIEW OF LITERATURE
Phytochemical properties
Β- Anisaldehyde and β- sitosterol were isolated and elucidated
from carbontetrachloride and chloroform soluble portions of methanolic
extract of stem bark of Averrhoa carambola (Md. Masum Mia et al.,
2007).
On GC/MS analysis of chloroform soluble fraction of leaves, high
percentage of esters (45.06%) dominated by byranillin acetate (7.63%) in
addition to pentacosane(11.12%) were revealed. Similarly GLC of
saponifiable and unsaponifiable fractions of petroleum ether extracts of
both stem and leaves show that high percentage of unsaturated fatty acids
(31.87%) is present in leaves than in stem and TSFA in stem is more than
those present in leaves. Β- amyrin, stigmasterol and β- sitosterol
represented major constituents of unsaponifiable fraction (Tadros, S.H et
al., 2004)
The Total Phenolic Content (TPC) analysis of fruit of Averrhoa
carambola (star fruit) revealed that the acetone extracts contain highest
amount of total phenolics followed by ethanolic and methanolic extracts
(C.F. Yap et al., 2009)
9
Volatile constituents present in methaylchloride extract of star
fruits showed the presence of methyl anthranilate (cause aroma), ethyl
sorbate, ethyl butyrate, 6-methyl-5-hepten-2-one, , 6-methyl-5-hepten-2-
ol, phenyl ethylacetate, limonene, phenylethyl alcohol,
etc.(Wilson,C.W.,et al.,1985)
Polyphenyloxidase was isolated from starfruit causing browning of
fruits(Tengku Adnan,T.A.B., 1986). Three insoluble Fibre-Rich Fractions
(FRFs) including insoluble dietary fiber, alcohol-insoluble solid and
water-insoluble solid isolated from star fruits could retard glucose
diffusion, postpone release of glucose from starch and inhibit the activity
of α –amylose to different extent (Chi-Fai Chau et al., 2003).
Alkyl phenols like 2,5-dimethoxy-3-undecylphenol and 5-
methoxy-3- undecylphenol and benzoquinones like 5-o-methylembelin
and 2-dehydroxy-5-o-methylembelin were isolated and elucidated from
the wood of Averrhoa carambola.
Another member of the family Oxalidaceae, Oxalis erythrorhiza
contain 3-heptadecyl-5-methoxy phenol and embelin in its
leaves.(Gabriela Egly Feresin, et al., 2003). Similarly purple leaves of
Oxalis triangularis contain anthocyanins like Mavidin-3-o(4-o-malonyl-
α-rhamnopyranosyl)-β-glucopyranoside)-5-o-β-glucopyranoside(Torgils
Fossen, et al.,2005).
10
Three c-glycosylflavanones (isovitexin, isoorientin and swertisin)
were present in the leaf extract of Oxalis corniculata(Hiroki mizokami, et
al., 2007).
Antimicrobial properties
Carbon tetrachloride soluble fraction and petroleum ether soluble
fraction of methanolic extract of bark of Averrhoa carambola showed
moderate inhibitory growth while chloroform solvent showed inhibitory
zone in growth of E.coli. Similarly Vibrio mimicus show moderate
inhibitory growth in petroleum ether extract and very low inhibitory
growth in chloroform and carbon tetrachloride solvents (Md.Masum
Mia,et al., 2007).
Klebsiella sp. and Staphylococcus aureus possess inhibitory zones
of growth in aqueous extract of stem of Averrhoa carambola
(B.Sripanidkulchai,et al., 2002).
Ethanolic extract of leaf of Averrhoa bilimbi showed Minimum
Inhibitory Concentration (MIC) against Bacillus egaterium and E.coli
(Mackeen,M.M, et al., 1997).
Similarly methanolic extract of leaf of Averrhoa bilimbi showed
less potent antimicrobial activity against Salmonella paratyphi and
Bacillus megaterium than fruit extract. Same is in the case of
11
Staphyllococcus aureus, Bacillus subtilis and Vibrio parahemolyticus
(Sreedam Chandra Das, et al., 2011).
Antioxidant properties
Methanolic extracts of star fruit exhibited high percentage
inhibition of DPPH radicals, Ferric Reducing Antioxidant Property(FRAP
value), antioxidant capacity and total Phenolics compared to aqueous
extracts and control after examining the effect of sonication treatment
(Annegowda,H.V., et al., 2007).
Ethanolic extract of star fruit showed a moderate potency in
reducing lipid peroxidation (37.08%) and increased the antioxidants-
TAA, SOD CAT, GsH and GPX ( Rupal, A.V., et al., 2011).
Anthelmintic properties
Averrhoa carambola showed anthelmintic activity in dose
dependent manner giving shortest time of paralysis and death with 100
mg/ml concentration (Anisha.N.Shah, et al., 2011).
Pharmacologic and medical studies
Averrhoa carambola tea extract show a very alternation of cell
cycle of Allium cepa,L., but neither induced an increase in number of
12
chromosomal damage in bone marrow cells of Wistar rats nor alter their
cell cycle (Pimenta Vincentini V.E., et al., 2001).
Aqueous leaf extact of Averrhoa carambola depresses Guinea pig
atrial inotropism due to reduction of L-type calcium current
(Vasconcelos,C.M.L., et al., 2008).
The leaves from carambola have been utilized for the production of
herbal medicine Glico-Vitae (GV) indicated for non-insulin dependent
Diabetes mellitus (NIDDM)(Marilene Provasi, et al., 2001).
Star fruit intoxication can occur in patents with moderate Chronic
Renal Impairment (CRI) resulting in hiccups, mental confusions, coma,
etc. (Alexandre Herbland, 2007)
Neurotoxic fraction obtained from star fruit specifically inhibited
GABA binding (Carolino,R.O.G, et al., 2005).
Oxalate may play a key role in star fruit neurotoxicity in
nephrectomized rat and prpbably in uremic patients (Hua-Chang Fang, et
al., 2007)
Aqueous extract of Averrhoa bilimbi controlled rise in glucose in
blood of mice that has induced Diabetes (Giuseppina Negri, 2005). It also
prevents platelet aggregation and thrombosis in blood vessels of
13
streptozotocin- induced diabetic rats (Pushparaj, et al., 2001).
Amentoflavone, a biflavanoid from Biophytum sensitivum could
inhibit the process of angiogenesis (Guruvayoorappan,C, et al., 2009).
Decoction of whole plant of Biophytum pertersianum has a
significant antagonistic effect on caffeine- induced calcium release from
sarcoplasmic reticulatum and hence can be used as antihypertensive
(Mouzou,A.P, et al., 2010).
Cytotoxicity
Oral administration of Averrhoa carambola juice induced serum
ALT (ALanine aminoTransferase) level an increase of 38% for rats
(ZyKhoo, CC Teh, et al., 2010).
Alcoholic extracts of stems of Averrhoa carambola exhibited
selective activity against U251 (brain tumour cell line) and that of leaves
was effective against Hepg2 (liver carcinoma cell line) only (Tadros,S.H
and Sleem,A.A., 2004).
Economic importance
 Used for making fresh juice, wines, etc.
 Ingredient in Antioxidant Green Tea as a Free Radical Fighter.
 Juice as a Beverage with high phytochemicals / phytonutrient
14
values: Vitamin C, Antioxidants, flavanoids, phytoferols etc.
 As a cosmetic ingredient especially in moisturicing and anti-ageing
creams
 Used to make beauty soaps in Thailand.
 Cleaning: The acid type carambola dissolves tarnish and rust,
occasionally used for cleaning and polishing metal.
 Stain remover:, fruit juice is used in washing clothes and to remove
spots and stains. Contains potassium oxalate which is used for dyeing
(stuartxchange.org).
15
MATERIALS AND METHODS
3.1. Plant material
The material was collected from the compounds of Napier
Museum, Thiruvananthapuram. It was identified using Flora of the
Presidency of Madras (J.S.Gamble, 1915). The plant material was
authenticated at the Herbarium of Botany Department, University
College, Thiruvananthapuram were voucher specimen (no. 2316) was
deposited.
3.2. Preparation of extract
The phytochemicals present in the plant material was extracted by
distillation method using soxhlet apparatus with methanol as solvent.
About 700 g of plant material i.e., leaves were collected and washed well
to remove dirts. Moisture was removed by blotting with a cotton cloth
and shade dried for 10 days. The dried leaves were powdered and 50 g of
powder sample was packed in a thimble and kept in soxhlet apparatus.
About 300 mL of methanol was poured into the bottom flask of soxhlet
apparatus. The whole apparatus was kept over a heating mantle and was
continuously heated for 8 hours at 650 C (Boiling point of methanol). The
extract was concentrated to dryness and the residues were transferred to a
pre-weighed sample bottle and were stored in a desiccator for further
16
studies (Harborne, 1973).
3.3. PHYTOCHEMICAL SCREENING OF PLANT EXTRACT
Different biochemical parameters like reducing sugar, flavonoids,
steroids and terpenoids, tannins, caumarins, saponins and glycosides were
analysed.
3.3.1. Qualitative Analysis
Test for reducing sugars (Fehling’s test)
The aqueous methanolic extract was added to boiling Fehling’s
solution in a test tube. A brick red colour indicates the presence of
reducing sugars.
Test for flavonoids (Shinoda test)
Take 1 mL of extract and add a few magnesium turnings, followed
by addition of conc. HCl drop by drop. Development of pink colour
indicates the presence of flavonoids.
Test for saponins
5 mL of the extract was shaken with 5 mL of distilled water and
heated to the boiling point. Froathing indicates the presence of saponins.
17
Test for alkaloids
To 5 mL of warmed extract add 10 mL 2% HCl, warmed and
filtered. 1 mL of aliquot was treated with a few drops of Dragendroff’s
reagent. Orange brown precipitate indicates the presence of alkaloids.
Test for steroids and terpinoids
In 2ml of dry and chloroform dissolved extract, add a few drops of
acetic anhydride and conc.H2SO4 and keep undisturbed for few minutes.
Formation of reddish brown colour indicates the presence of steroids
while pink colour indicates the presence of terpinoids.
Test for Tanins
To 1 mL of the extract, add 2 drops of 5% FeCl3. Presence of dirty
green indicates the presence of tannins.
Test for coumarins
1 mL of the extract was dissolved in methanol followed by the
addition of a few drops of alcoholic NaOH. Conc.HCl was added through
the sides of the test tubes which shows the appearance and disappearance
of yellow colour, indicating the presence of coumarins.
Test for Glycosides (Killer-Killani test)
To 0.5 g of the extract diluted to 5 mL with distilled water, add 2
mL of glacial acetic acid followed by a drop of Ferric chloride solution.
18
This was underplayed with 1 mL of H2SO4. A brown ring at the interface
indicates the presence of glycosides.
3.3.2. Identification of some compounds by HPLC (High
Performance Liquid Chromatography)
Apparatus and Chromatographic conditions
A supercritical fluid extractor SFE-2 (Applied Separation, USA)
which is capable of pressure up to 680 bar and temperature up to 240ºC,
static and dynamic extraction with flow from 0 to 10 L/min (gaseous
carbon dioxide) and extraction vessels from 5 ml to 1l were used. An
Agilent 1200 liquid chromatograph system (Agilent technologies, CA,
USA) consisting of binary pump, an auto-sampler and diode-array
detector was used.
The column configuration consisted of an Agilent Zorbax Extend
reversed-phase C18 column (250 mm × 4.6 mm, 5 μm). Detection
wavelength was set at 220 nm. The mobile phase consisted of A
(methanol) and B (deionized water), using a linear gradient: 0-40 min
(85% A), 40-60 min (85% A-95% A). The flow rate was 1.0 ml/min. The
column temperature was maintained at 30 ºC.
19
Preparation of standard solutions - Averrhova
A mixed standard stock solution containing
a) Epigallocatechin
b) Catechin
c) Caumarin
d) Squamostatin
e) Squamocin
f) Ascorbic acid
g) Gallic acid
h) Gallotannin
i) Abscisic acid
j) Vomifoliol
k) Proanthocyanidin
l) Tocopherol (m)nerol
m) Linalool
n) Geraniol
o) Terpenol
p) Roseoside
q) Methyl benzoate
20
Was prepared in methanol. Working standard solutions were
prepared by diluting the mixed standard solution with methanol to give
six different concentrations within the ranges:
a) 3.8-45.6 μg/ml
b) 2.1- 6.7μg/ml
c) 3.2-36.4 μg/ml
d) 4.5-51.7 μg/ml
e) 2.7-29.2 μg/ml
f) 0.58-17.3 μg/ml;
g) 2.5-18.9 μg/ml
h) 4.3-48.9 μg/ml
i) 3.3-41.8μg/ml
j) 12.7-29.2 μg/ml
k) 10.58-17.3 μg/ml
l) 12.5-18.9 μg/ml;
m) 14.3-28.9 μg/ml
n) 33.3-41.8μg/ml
o) 22.7-29.2 μg/ml
p) 12.58-17.3 μg/ml
q) 2.5-18.9 μg/ml
r) 44.3-48.9 μg/ml
21
For calibration curves. The standard solutions were filtered through
a 0.45 μm membrane prior to injection. The standard stock and working
solutions were stored at 4°C.
Preparation of sample solutions
The extract sample of Averrhova carambola under optimized
conditions (extraction pressure: 30 Mpa; extraction temperature: 35ºC;
extraction time: one hour; 20 ml 95% ethanol modifier) was used.
After evaporating ethanol to dryness by a rotary evaporator, residue
was dissolved in methanol in a 25 ml flask, and then filtrated through a
0.45 micro-m millipore filter before HPLC injection. Three aliquots of
the solution (20 μl) were injected to RP-HPLCDAD system. The graph
obtained after the elution of compounds was compared with the standard
graph and major compounds were detected.
3.3.3. Quantitative analysis
Determination of moisture content
5 g of material was taken in a preweighed petridish. The petridish
was placed without lid into an oven at 1100C for 3 hours. The petridish
was taken out and closed immediately with alid. The dish was then
cooled in a dessicator and weighed. The amount of moisture was
calculated from the difference in weight.
22
Estimation of Total Carbohydrate
Weighed amount of fresh tissue was homogenized with distilled
water. The homogenate was filtered using a two layered cheese cloth. The
filtrate was then centrifuged at 10,000 rpm for fifteen minutes. The
supernatant was collected and the volume was made up to 25 mL using
distilled water. An aliquot of sample was pipette out and 4 mL Anthrone
reagent was added. It was then kept in a boiling water bath for ten
minutes. The tubes were cooled and the absorbance was measured at 530
nm in a spectrophotometer. The amount of total carbohydrates was
determined using the standard graph of glucose (Hedge.J.E., et al., 1962).
Estimation of Protein
Total protein present was estimated by Lowry method. (Lowry.
O.H., et al., 1951).one gram of fresh leaf tissue was ground thoroughly in
10 mL Phosphate Buffer. The homogenate was filtered through a double
layered cheese cloth and centrifuged at 5,000 rpm for 10 minutes. The
supernatant collected was made to a known volume of 10 mL using
Phosphate buffer. An aliquot of one mL was taken and 10%
TriChloroAcetic acid (TCA) was added and shaken well. This was kept
under refrigeration for fifteen minutes and then centrifuged at 10,000 rpm
for ten minutes. The upper layer was decanted; the pellet collected and
dissolved in 10 mL 0.1 N NaOH solution. 0.1 mL aliquot was pipette out
23
and made up to 1mL by using 0.1 N NaOH. To this 5 mL of reagent C
(mixture of 2g Na2CO3 in20 mL 0.1N NaOH and 0.25 g
KNaC4H4O6·4H2Oin 25 mL distilled water in 50:1 ratio) was added and
kept for ten minutes. Then add 0.5 mL of alkaline Folin’s reagent and
kept for thirty minutes. Absorbance was measured at 670 nm. The protein
present was calculated from the standard graph of Bovine Serum
Albumin (BSA).
Estimation of Total Phenols
Weighed amount of fresh tissue was chopped and putinboiling
methanol (80%) for ten minutes and refluxed. The refluxed matter was
homogenized. The homogenate was filtered and centrifuged at 10,000
rpm for ten minutes. The supernatant was collected and the volume was
made upto 20 mL using 80% methanol. 0.5 mL Folin-Ciocalteau reagent
followed by 2 mL 20% Sodium carbonate was added to an aliquot of
sample and mixed thoroughly. The reaction of phenol with
phosphomolybdic acid in Folin-Ciocalteau reagent in alkaline medium
produced a blue coloured complex. The tube was kept in a boiling water
bath for 1 minute, cooled, centrifuged and the supernatant was taken to
measure the absorbance at 650 nm. The amount of Total Phenols was
determined using a standard graph of catechol (Malick.C.P., et al., 1980).
24
Estimation of Reducing sugars
1 g of fresh leaf tissue was taken and homogenized in 10 mL
distilled water. The homogenate was filtered using a cheese cloth. The
filtrate was centrifuged at 10,000 rpm for ten minutes. The supernatant
collected was made upto a known volume of 10 mL. About 0.2 mL 0f this
was taken and made upto 1 mL using distilled water. 2 mL DNS
(DiNitroSalicylic Acid) reagent was added and kept in a boiling water
bath for ten minutes. This was later cooled to get an orange red colored
solution. Absorbance was measured at 540 nm (Plummer, D.T., et al.,
1987).
Estimation of Amino acids
Fresh tissue was weighed to 1 g and refluxed in 80% methanol for
ten minutes. This was grinded thoroughly in a mortar and pestle and
filtered out. The filtrate was centrifuged at 7500 rpm for ten minutes. The
collected supernatant was made up to a known volume of 10 ml using
80% methanol. 0.1 mL of this was taken and 5 mL of Ninhydrin reagent
was added. It was shaken well and heated in a boiling water bath for ten
minutes. The tubes were cooled under running tape water and the
absorbance was measured at 570 nm. The amount of amino acids was
estimated using the standard graph of leucin (Moose. S.,et al., 1948).
25
Estimation of Chlorophylls
Ig of fresh leaf tissue was grinded in 10 mL 80% acetone. The
filtrate obtained through cheese cloth was centrifuged at 5,000 rpm for
five minutes. The supernatant was collected and made upto a known
volume of 10 mL. From this 1 ml was taken and made upto 5 mL using
80% acetone. The absorbance was measured at 645, 663, 652 and 490 nm
for calculating the amount of Chlorophyll a, b, total Chlorophyll and
Caroteinods respectively (Arnon, D.I, 1949).
26
3.4. ANTHELMINTIC ASSAY
The Assay was performed in Pheretima posthuma due to its
anatomical and physiological resemblance with the intestinal round worm
(Ascaris lumbricoides) parasite of human beings (Thorn, G.W., et al.,
1977).
3.4.1. Worm collection
The Indian Earthworm Pheretima posthuma (Annelida) was
collected from Botanical garden, Department of Botany, University
College, Thiruvananthapuram.
3.4.2. Anthelmintic activity
The anthelmintic assay was carried as per the method of
Anisha.N.Shah et al., 2011 with necessary modifications. The leaves of
Averrhoa carambola were dried, ground to coarse powder and boiled.
The aqueous extract obtained was filtered and concentrated by
evaporation to get extract.
3.4.3. Preparation of test sample
Samples for in vitro study were prepared by dissolving 2.5 g of
extract in 25 mL of distilled water to obtain stock solution of 100 mg/mL.
From this stock solution, different working dilutions were prepared to get
27
a concentration range of 10, 50 and 100 mg/mL.
50 mL formulations containing three different concentrations each
of extract (10, 50 and 100 mg/mL in distilled water) were prepared and
two worms were placed in it. Time for paralysis was noted when no
movement of any sort could be observed except when the worms were
shaken vigorously. Time of death of worms was recorded after
ascertaining that the worms neither moved when shaken vigorously or
when dipped in warm water (500C). Albendazole (10, 50 and 100 mg/mL)
was used as reference standard while distilled water as the control.
3.5. ANTIMICROBIAL STUDIES
3.5.1. Sterilization of glass wares
The petridishes, test tubes and other glasswares were washed with
a mild detergent and rinsed with sterile distilled water. Then they are
autoclaved at 1210C for ten minutes.
3.5.2. Preparation of Peptone water
The media was prepared by taking 13.6 g of Peptone powder in
1000 mL distilled water. Then it is poured in 5-6 mL in each test tube.
The test tubes were then autoclaved at 121 0C for twenty minutes and
stored.
28
3.5.3. Preparation of media for culturing bacteria
The media used for culturing bacteria was nutrient agar medium. It
was prepared by adding 28 g of nutrient agar in 1000 mL of distilled
water taken in a beaker. For solidification of the medium, 10 g of agar
was dissolved by placing it in a boiling water bath. Then it was
autoclaved at 1200C for fifteen minutes. Required volume of molten
medium were poured into sterile petridishes under aseptic condition and
stored. The bacterial strains used were E.coli, Enterococcus sps.,
Staphylococcus and Bacillus pyocyaneus. The pure samples were
obtained from the Department of Microbiology, Govt. Medical College,
Thiruvananthapuram.
3.5.4. Inoculation of bacteria
Inoculation was done under flame in highly aseptic condition under
laminar airflow chamber. The pure culture obtained from laboratory was
taken and from that 1-3 spores of bacteria were transferred to 5 mL of
Peptone water in the test tube. Then it was incubated for one hour at 370C
for regeneration. After one hour, this was taken out and swabbed into the
nutrient agar plate. Then it was kept aside.
3.5.5. Preparation of Disc for inhibition studies
In the disc diffusion method, discs of 4 mm diameter were cut out
from Whatman No.42 filter paper. They were sterilized by autoclaving at
1200C for fifteen minutes. Then it was stored in aseptic condition in a test
29
tube. During the time of treatment, the discs were taken out from the test
tube with the help of a foreceps. The crude extract of different
concentrations were pipetted out and poured into different sterile
petridish. The concentrations were 1mg/mL, 2mg/mL and 3 mg/mL of
each extract. Then 10 filter paper discs were placed into each petridish.
Cover it with a glass lid and kept for one hour so that the filter paper will
be fully saturated with crude extract.
With the help of forceps, one disc from each petridish of various
concentrations, which are saturated with crude extract, was taken. These
discs were placed on each petridish containing bacterial spores. A control
disc was treated with the solvent, methanol. The standard disc used here
is an antibacterial drug, Ampicillin. Then it was covered with petriplate
and was kept for incubation overnight at 370C. After one day, zone of
inhibition was noted only if the plant extract possess antibacterial activity
(Barry, A.L., 1980).
30
RESULTS AND DISCUSSION
4.1. PHYTOCHEMICAL SCREENING
4.1.1. Qualitative analysis
The qualitative analysis of leaf extract indicated the presence of
tannin, reducing sugar, Terpinoids, caumarin and saponin. But
glycosides, alkaloids, steroids and flavonoids are absent (Table 1).
4.1.2. Quantitative analysis
The qualitative analysis of fresh leaves of Averrhoa carambola, L.
showed the presence of Protein, Phenol, Amino acids, Reducing sugar,
Carbohydrate, Chlorophyll a, Chlorophyll b and caroteinods in different
amounts.
Proteins and amino acids are present in 6.92 and .078 mg/mL while
carbohydrate and reducing sugar are present in 147 and .029 mg/mL
respectively.
The pigments chlorophyll and caroteinoid are present in 4.55 and
0.015 mg/mL respectively. Among Chlorophylls, Chlorophyll a and
chlorophyll b are present in 3.64 and 3.19 mg/mL respectively.
31
Table.1
Name of compound Present/absent
Reducing sugar Present

Glycoside Absent
Alkaloid Absent
Terpinoids Present
Steroids Absent
Flavonoid Absent
Tannin Present
Caumarin Present
Saponin Present
32
Table. 2
Name of compound Amount (Mg/gL)
Protein 6.92
Phenol 0.005
Amino acids 0.078
Reducing sugar 0.029
Carbohydrate 147.0
Total chlorophyll 4.55
Chlorophyll a 3.64
Chlorophyll b 3.19
caroteinoid 0.015
moisture content 90.76%
33
The moisture content is about 90.76% and the amount of
Carbohydrate is the most and that of phenol is the least (Table 2).
Carbohydrates represent an important class of naturally occurring
substance. About 80% of the dry weight of green plants is made of
carbohydrates. Catabolism of carbohydrates provides the major share of
the energy requirement for life and performance of work, glucose is an
immediate source of energy in organism (Lehninger, 1995)
Proteins are the essential constituents of life which make up about
12% of protoplasm. They form enzymes, transport materials, regulation
of metabolism, movement and defense reactions (Lehninger, 1995)
Phenols are biologically active compounds. They are aromatic
compounds possessing one or more phenolic carboxyl group. Plants
respond to infection or injury by the accumulation of phenols. Phenols are
active against a wide range of microorganisms. It form the basic frame
work of lignin and also helps in photosynthesis by absorbing U.V. passed
on to the chlorophyll (Takayama, et al., 2001)
Chlorophyll and Caroteinoid are the major photosynthetic pigments
found in plants. Chlorophyll plays crucial role in the phenomenon of
photosynthesis. Chlorophyll a and Chlorophyll b are the common
chlorophyll pigments in green plants. Caroteinoids include Carotenes and
xanthophylls (Arnon, 1949).
34
4.1.3. Identification of compounds by HPLC
The HPLC method is adopted for the identification of compounds
present in the Methanolic extract of leaves. The compounds identified
were listed below (List.1). About 13 compounds were identified.
35
LIST-1
HPLC Compounds Identified – Averrhova
1. Gallic acid
2. Citric Acid
3. Epigallocatechin
4. Epigallo Catechin Gallate
5. Catechin
6. Epicatechin
7. L-Ascorbic Acid
8. Proanthocyanidin
9. Abcisic Acid
10. Vomifoliol
11. Roseoside
12. Methyl benzoate
36
4.2. Anthelmintic assay
The leaves of Averrhoa carambola displayed a significant
anthelmintic activity in dose depended manner giving shortest time of
paralysis (P) and death (D) with100mg/ml concentration as shown in
Table 3 (Fig 5, 6 and 7). The anthelmintic activity of aqueous extract was
comparable with that of standard drug at 100mg/ml concentration, plant
extract showed paralysis in 9.44 minutes and death in 16 minutes while
Albendazole exhibited similar effects at 8 and 18 minutes respectively
(Fig.7). Control worms were live up to 24 hours (Fig.8).
As the plant extracts are found to contain tannins, which are
Polyphenolic compounds, there is a possibility that their effects are like
synthetic phenolic anthelmintics e.g. - niclosamide, oxyclozanide,
bithionol etc. which are reported to interfere with energy generation in
helminth parasites by uncoupling oxidative phosphorylation(Martin, R.J,
1997). Another possible anthelmintic activity of tannins is that they can
bind to free proteins in gastrointestinal tract of host animal or
glycoprotein on the cuticle of parasite and may cause death(Thompson,
D.P. et al., 1995).
37
Table 3
Anthelmintic activity of Averrhoa carambola,L.
Time taken for Paralysis (P) and

Concentration Death (D) of Pheretima posthuma
Sample
(mg/ml) in minutes
P D
Albendazole 10 28.23±1.34 67.36±5.65
Albendazole 50 13.36±1.46 38.5±1.32
Albendazole 100 8.4±1.14 17.48±2.42
Plant extract 10 83.05±8.43 187.5±30.01
Plant extract 50 25.55±2.54 33.12±3.15
Plant extract 100 9.44±2.01 17.24±1.47
All values represent Mean ± SEM; n=2 in each group.
38
4.3. Antibacterial assay
The present study indicates that the extract from leaves showed
positive reaction against pathogens. E.coli, Enterococcus sps.,
Staphylococcus and Bacillus pyocyaneus.
In the present investigation, the soaked filter paper discs carrying
plant extract in methanol bring about an zone of inhibition of 1.9 mm,
2.1mm and 1.0 mm for 100mg/mL for E.coli , Enterococci sp. and
Bacillus pyocyaneus respectively. The zone of inhibition for 50 mg/mL
were 1.7 mm, 1.6 mm, 1.5mm, 1.9 mm for E.coli , Staphylococcus sp..,
Enterococci sp. and Bacillus pyocyaneus respectively. Similarly 1.5mm
and 1.6 mm were the zone of inhibition for E.coli , and Enterococci sp.
(Fig.9, 10,11 and 12), (Table.4).
The cell walls of gram negative bacteria were thinner compared to
that of gram positive bacteria. So they undergo lysis more easily than the
others. This makes them more sensitive to drugs than gram positive
bacteria.
The immediate reason for positive results may be due to the
presence of chelating agents and also due to the favorable change in the
permeability of cell membranes.
39
Table.4
Zone of inhibition of bacteria
Test organism Disc Conc.of Zone of Control Standard

no. extract inhibition (Methanol) (mm)
(mg/mL) (mm)
E.coli 1 100 1.9
2 50 1.7 - 6
3 10 1.5
Staphylococcus sp. 1 100 -
2 50 1.6 - 4.7
3 10 -
Enterococci sp. 1 100 2.1
2 50 1.9 - 3.6
3 10 1.6
Bacillus pyocyaneus 1 100 1.0
2 50 1.5 - 3.9
3 10 -
40
Thus from the present study, it was proved that Averrhoa
carambola has antibacterial property as found in previous works
(Md.Masum Mia, et al., 2007; Burgorn Sripanidkulchai. et al., 2002)
41
SUMMARY
Averrhoa carambola, L.of Family Geraniaceae is a tree widely
cultivated in Asian and African countries. The leaves were used for the
treatment of biliousness and diarrhea.
Leaves collected from Thiruvananthapuram were used to make
extract by using distillation method using soxhelet apparatus with
methanol as solvent. The weighed shade dried and powdered material
was continuously heated at the boiling point of solvent. Later it was dried
and transferred to a pre-weighed sample bottle.
Different biochemical parameters like reducing sugar, flavonoids,
steroids and terpenoids, tannins, caumarins, saponins and glycosides were
analysed. Quantitative test for plant like determination of moisture, total
carbohydrate, aminoacid, total protein, total phenol and chlorophyll were
done using standard methods.
The anthelmintic assay was done in Pheretima posthuma. Averrhoa
carambola showed anthelmintic activity. However dose and form in
which they can be used requires standardization. . Future scope involves
isolation of phytoconstituents responsible for activity. Pure cultures of
Gram positive - Enterococci sps. and Staphylococcus sps. And gram
negative- E.coli and Bacillus pyocyaneus were used for antibacterial
42
studies and zone of inhibition were observed high in crude extract
(100mg/mL).
Thus from the present study, it was made clear that Averrhoa
carambola, L. is not only a food source, but also possess very effective
anthelmintic and antibacterial properties. Thus the economic potentialities
of this plant may be exploited in future, after more extensive studies.
43
REFERENCE
 Alexandre Herbland, et al., Star fruit poisoning is potentially life
threatening in patients with moderate chronic renal failure, Journal
of intensive care medicine, 2007, vol.35(8);1459-63
 Anisha N. Shah, Anuja B. Raut, Prof. Akhay Bahetiand Dr.
Kuchekar, B.S., Invitro anthelmintic activity of leaf extract of
Averrhoa carambola against Pheretima posthuma,
Pharmocologyonline, 2011;524-27
 Annegowda H.V and Mansor, S.M, Influence of Sonication
treatments and extraction solvents on the phenolics and antioxidants
in star fruits, Fruits, vol.59(6), 2009; 256-58.
 Arnon, D.I., Plant Physiol.24, 1949.
 Bailey, H.S., Manual of cultivated plants, MacMillan Company,
NewYork, 1948; 601-3.
 Barry, A.L., Procedures for testing antimicrobial agents in agar
media. In: Antibiotics in Laboratory Medicines, Williams and
Wilkins Co. Baltimore, USA, 1980.
44
 Chi-Fai Chau, Chien-Hung Chen, Ching-Yi Lin, Insoluble fiber-
rich fractions derived from Averrhoa carambola: hypoglycemic
effects determined by in vitro method.
 Carolino, R.O.G., et al., Convulsant activity and neurochemical
alternations induced by a fraction obtained from the fruit Averrhoa
carambola (Oxalidaceae: Geraniales), Neurochemistry international,
2005, vol.46(7); 523-31.
 Deore S.L, Khadbadi S.S., In vitro anthelmintic studies of
Chlorophytum borivilianum sant. &Fernandez tubers, Indian
Journal of Natural Products and Resources 2010; 1:53-5.
 Facciola, Stephen. Cornucopia: a Source Book of Edible Plants.
Kampong Publications, 1990. p. 39.
 Gamble J.S., Flora of the Presidency of Madras, vol.1, Bisen
Singh, Mahendrapal Singh Pub., Dehradun,1915; 130-33
 Giuseppina Negri, Diabetes mellitus: plants and natural active
principles, Journal of Pharmaceutical Sciences, 2005, vol.41.
 Guruvayoorappan, C. and Kuttan, K., Inhibition of tumor specific
angiogenesis by amentoflavone, Journal of Biochemistry, 2010,
vol.73 (2); 209-18.
 Hedge, J. E. and Morfreiter, B.T., In carbohydrate chemistry, (Eds.
45
Whistter, R.L. and Be Miller, J.N.), Academic Press, New York,
1962.
 Hiroki Mizokami, Kaori Tomita Yokotani and Kunijiro Yoshitama,
Flavonoids in the leaves of Oxalis corniculata and sequestration of
the flavonoids in the wing scales of the pale grass blue, Journal of
Plant Research, 2007, 121(1); 133-6.
 Hua-Chang Fang et al., The role of oxalate in star fruit
neurotoxicity of five-sixths nephromized rats, Journal of Food and
Chemical Toxicity, 2007, vol.45(9); 1764-69
 Jain.S.K, Medicinal Plants, Aphsa Pub. Ltd., New Delhi; 1987;
1-4.
 Lehninger, A.L., Biochemistry, KAlyani Publishers, New Delhi,
1995.
 Lewis et al., Medicinal botany- plants affecting health, 8112
 Lowry, O.H, Rosebrough,N.J., Farr,A.l. and Randall, R.J., J.Biol.
Chem., 1951, 193; 265.
 Mackeen,M.M., Ali, A.M., Salleh, K.M.,and Kawazu, Antimicrobial
and cytotoxic properties of some Malaysian Vegetables (Ulam),
Journal of Pharmaceutical Biology, 1997, 35(3); 174-8
46
 Mallick, C.P. and Singh, M.B., In plant enzymology and Histo
enzymology, Kalyan Publishers, New Delhi, 1980; 286.
 Marilene Provasi, et al., Evaluation of the toxicity and
antihyperglycemic potential of Averrhoa carambola L.
(Oxalidaceae), Acta Scientiarum, Health Sciences, 2001, vol.23.
 Martin, R.J. Mode of action of anthelmintic drugs, Vet.J. 1997,
154; 11-34.
 Md. Masum Mia, Mohammad S. RAhman, Kohinur Begum, Bikis
Begam and Mohammad Rashid, phytochemical and biological
stidies of Averrhoa carambola, Dhaka Univ. J. Pharm. Sci. 6(2):
125-128, 2007 (December).
 Menon, I A and H F Haberman, Dermatological writings of ancient
India Medical History. 1969 October; 13(4): 387–392.
 Moose,S and Stein,W.H., In the methods in Enzymol. (Eds.
Colwick,S.P. and KAlpan,N.D.),1948.
 Morton, Julia F. Fruits of Warm Climates. Creative Resources
Systems, Inc. 1987. pp. 125-128.
 Mouzou, A.p et al., Effects of the Biophytum petersianum
(Oxalidaceae) decoction on calcium release from the sarcoplasmic
reticulatum in the skeletal muscle cells, Phytotherapie, 2009,
vol.8(4); 231-35.
47
 National Bureau of Plant Genetic Resources (NBPGR) Annual
Review Booklet, 2006.
 Pimenta Vincentini V.E., Camporato, M.L., Teixeira, R.O and
Mantovani, M.S, Averrhoa carambola L., Syzygium cumini (L.)
Skeels and Cissus sicyoides L.; medical herbal tea effects on
vegetal and animal test systems, Acta Scientiarum, Maringa,
v.23(2), 2001; 593-8.
 Plummer,D.T, An introduction to Practical biochemistry, Tata
McGraw Hill Pub. Co. Ltd, New Delhi, 1987; 180-1.
 Pushparaj,P.N., Huattan, B.K., The mechanism of hypoglycemic
action of the semi-purified fractions of Averrhoa bilimbi in
Stereptozotozin-diabetes rats, Life Sci, 2001,vol.70; 535-47
 Raju VK; Susruta of ancient India; Indian J Ophthalmol 2003;
51:119-22.
 Rupal, A.V. and Narasimhacharya, A.V.R.L., Antioxidant potential
of seasonal fruits in fluoride toxicity: an in vitro study, Journal of
Cell and Tissue Research, 2011, vol.11 (2); 2789-94.
 Sharma. G.R.K., Trivedi. P.C.; Medicinal plants, Traditional
Knowledge; J.K. International Pub. House Pvt. Ltd, New Delhi;
2006; 118-21.
48
 Sreedam Chandra Das et al., Antibacterial and cytotoxic activities
of methanolic extracts of leaf and fruit parts of the plant Averrhoa
bilimbi (Oxalidaceae), American Journal of Scientific and
Industrial Research, 2011 2153-649x.
 Sripanidkulchai,S., et al., Anti-inflammatory and Bactericidal
Properties of Selected Indigenous Meicinal Plants used for
Dysuria, J.Pharm.Sci.,2002,26(1-2): 33-8
 Tadros, SH TS El Alfy and TA Ibrahim ,Volatile Constituents,
Nutritive Value and Bioactivity of the Pericarp of
Averrhoa carambola L. Fruits Grown in Egypt - Bulletin of the
Faculty of Pharmacy , 2004.
 Takayama, et al., Journal of natural products, 2001, 64(9), 1224-5.
 Tengku Adnan, T.A.B., Augustin, M.A. and Ghazali, H.M.,
Polyphenoloxidase from star fruit (Averrhoa carambola L.), J. Sci.
Fd. Agric. 36: 1259- 65.
 Thorn G.W., Adams R.D., Braunwald E., Isselbacher K.J. and
Petersdorf P.G; Harrison’s Principles of Internal Medicine,
McGraw Hill Co., New York, 1977: 1088.
 Thompson, D.P., and Geary, T.G., The structure and function of
helmintic surfaces. IN Marr, J.J. eds. Biochemistry and molecular
Biology of parasites, Academic Press, New York, 1955; 203-32.
49
 Tiwari, D.N., Medical Plants an Introduction, Department of Indian
system of Medicine and Homeopathy.
 Thomas S., Patil D.A., Pharmacognostic Evaluation and
Physicochemical Analysis of Averrhoa Carambola L. Fruit,
Journal of Herbal Medicine and Toxicology 2008; 2 : 51-54.
 Torgils Fossens et al., Acylated anthocyanins from leaves of Oxalis
triangularis, Journal of Phytochemistry, 2005, 66(10); 1133-40.
 Vasconcelos,C.M.L., et al., Aqueous leaf extract of Averrhoa
carambola L. (Oxalidaceae) reduces both the inotropic effect of
BAY K 8644 on the guinea pig atrium and the calcium current on
GH3 cells, Brazilian Journal of Pharmacognosy,2008, 18(4);
539-43.
 Wilson, C.W., Shaw, P.E., Knight, R.J., Steven Nagy and Marty
Klim, Volatile constituents of carambola(Averrhoa carambola L.)
J. Agric. Food Chem. 1985,33; 199-201
 Yap, C.F., Ho,C.W.,et al., Optimization of extraction conditions of
Total Phenolic Compounds from Star fruit(Averrhoa
carambola.L.) Residues, Sains Malaysiana 38(4)2009; 511-20
 Yusuf, V.K. and Sinohin, V.O., Cananga odorata(Lamark) Hook.f,
Plant resourses of South East Asia, 1999; 70-4.
50
 ZyKhoo,CC Teh, et al., Effect of Averrhoa carambola juice in the
serum ALT (ALanine aminoTransferase) level for rats, Seizure,
2010, vol.14(7), 521-25.
Online reference
 http://www.thefruitbook.com/fruits/FINFO/carambola.html
 http://www.rarefruit.hypermarket.net/exotic3.html
 http://www.agroforestrytreedatabase.html
 http://www.stuartxchange.org/Balimbing.html
51

Averrhoa Carambola, L. Leaf Extract: Raju V.K., 2003

Uploaded by

Copyright:

Available Formats

Averrhoa Carambola, L. Leaf Extract: Raju V.K., 2003

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Averrhoa Carambola, L. Leaf Extract: Raju V.K., 2003

Uploaded by

Copyright:

Available Formats

Averrhoa carambola, L.

Medicinal plants, the World’s oldest known healthcare products,

plants and other natural resources (G.R.K.Sharma, 2006).

The history of medicine dates back perhaps to the origin of human

race. Rig veda written between 4500-1600 BC is the oldest repository of

human knowledge in India. Later, Suhsruta Samhita and Charaka Samhita

184 chapters and description of 1120 illnesses, 700 medicinal plants, a

detailed study on Anatomy, 64 preparations from mineral sources and 57

preparations based on animal sources (Raju V.K., 2003). The Charaka

Samhita was recorded several thousand years ago from the

Teachings of the sage Punarvasu Atreya. The Charaka Samhita is

metrical verses, which are often committed to memory, in toto, by

modern medical students of Ayurveda. It presents most of the theoretical

edifice of Ayurveda and concentrates on the branch of Ayurveda called

Traditional healthcare systems using medicinal plants can be

recognized and used as a starting point for the development of novelties

in drugs. The volatile components of medicinal plants and herbs are

extracted that can be used for various purposes.

for medicinal purposes (NBPGR Booklet, 2006). Overall 119 plant

derived prescription drugs are commonly used in different countries, 74%

of which were discovered due to chemical isolation of active compounds

of plants used in traditional system of medicine (Sharma,G.R.K., 2006).

India is ranked no.8 in World Biodiversity. About 70% of India’s

alpine areas. Macrostudies showed that a large percentage of known

medicinal plants occur in the dry and moist deciduous vegetations as

compared to the evergreen or temperate habitats (Tiwari, 2000).

In some countries like India, pharmaceutical companies are already

marketing preparations of tablets, capsules, etc. they are made directly

Hepatitis. Medicinal plants and herbs contain substances known to

modern and ancient civilizations for their healing properties. Until

compounds in the nineteenth century, medicinal plants and herbs were

sole source of active principles capable of curing man’s aliments. They

continue to be important to people that do not have access to modern

medicines and moreover, modern pharmaceuticals rely heavily on the

same active principles (Yusuf et al., 1999).

Drug discovery, ethnobotany and traditional and indigenous

for medicinal plants have been discovered and popularized, sustainability

has become increasingly an issue; concern over the growth in biopyracy

and habitat (Lewis, et al., 2003).

The present study is concentrated on Averrhoa carambola, L. of

the family Geraniaceae. This plant is most common in African countries

it is a native species, is less common among people as a food source. But

fibers, protein, Vitamin C, Potassium, etc.(Facciola, et al., 1990). The

phytochemical screening as well as antimicrobial and anthelmintic assays

Current name: Averrhoa carambola (The generic name is after

Averrhoes (1126-98), the widely known Arab Philosopher. The specific

early by the Portuguese).

According to Bentham and Hooker’s classification

Averrhoa acutangula., Stokes

1.2. Common names

Malayalam : Chaturappuli, Irumbanpuli, Karappola

Hindi : kamrakh, kamranga

English : carambola, star fruit, Carambola apple, Karazola,

French : blinblin longue, carambolier, cornichon du pays

Indonesian : belimbing manis

Malay : belimbing manis

Tree to 30 ft (Fig.1); young parts finely pubescent; leaf 5-11,

flowers in the leaf axils(Fig.2); petals about ¼ in. long, white-marked

sweet when ripe, fragrant (Fig.3&4)(Bailey,L.H., 1948)