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Composition of Water Extract from Wild Bitter Gourd (Momordica charantia L.) Fruit for application as Antifeedant and Mortality Test on Armyworm Larvae (Spodoptera litura Fab.)

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This study was to determine the fruit maturity level of wild bitter gourd and the composition of water extract which is effective as antifeedant and appropriate for mortality test on armyworm larvae. Results obtained show that water extract of wild bitter gourd fruit contains phenolic compounds, flavonoids and triterpenoids. Terpenoid compounds contained in water extract of fruits at maturity level 4H, 8H and 12H were momordicoside L, momordicoside K, compound 3β, β7, 25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2= H and momordicine 1.Fruit maturity level 4H 50% and 4H 60% resulted to the highest antifeedant index, ie 40.08% and 44.20%.LC50at fruit maturity level 4H observed on day 7 was 40%.
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Composition of Water Extract from Wild Bitter Gourd
(Momordica charantia L.) Fruit for application as
Antifeedant and Mortality Test on Armyworm Larvae
(Spodoptera litura Fab.)
Tri Wardhani (Corresponding author)
Doctoral Student of Dept. of Agricultural, Brawijaya University
Work at Dept of Agricultural, Widyagama Malang University
Jl. Borobudur 35, Malang, Indonesia
Tel: 62-857-5541-1330 E-mail: twd@widyagama.ac.id
Abdul Latief Abadi
Dept. of Agricultural, Brawijaya University
Jl. Veteran, Malang, Indonesia
Tel: 62-818-384-040 E-mail: latiefabadi@ub.ac.id
Toto Himawan
Dept. of Agricultural, Brawijaya University
Jl. Veteran, Malang, Indonesia
Tel: 62-817-910-3300 E-mail: totohimawan@yahoo.co.id
Aulanni’am
Dept. of Mathematics and Natural Science, Brawijaya University
Jl. Veteran, Malang, Indonesia
Tel: 62-812-331-7600 E-mail: aulani@ub.ac.id
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Received: June 16, 2015 Accepted: July 4, 2015
doi:10.5296/jbls.v6i2.8084 URL: http://dx.doi.org/10.5296/jbls.v6i2.8084
Abstract
This study was to determine the fruit maturity level of wild bitter gourd and the composition
of water extract which is effective as antifeedant and appropriate for mortality test on
armyworm larvae. Results obtained show that water extract of wild bitter gourd fruit contains
phenolic compounds, flavonoids and triterpenoids. Terpenoid compounds contained in water
extract of fruits at maturity level 4H, 8H and 12H were momordicoside L, momordicoside K,
compound 3β, β7, 25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2= H and
momordicine 1. Fruit maturity level 4H 50% and 4H 60% resulted to the highest antifeedant
index, ie 40.08% and 44.20%. LC50 at fruit maturity level 4H observed on day 7 was 40%.
Keywords: Wild bitter gourd, Water extract, Antifeedant, Mortality, Triterpenoids,
Armyworm larvae
1. Introduction
Armyworm (Spodoptera litura Fab.) is a polyphagous insect, which has many kinds of host
plants. Armyworm is an important pest in many kinds of plants, such as weeds like grinting
and reeds, horticultural crops such as tomatoes, chili, beans, cabbage, onions, spinach, kale,
potatoes, crops such as rice, corn and soybeans, as well as plantation crops such as citrus,
cotton, sugarcane. Armyworm can reduce the production of soybean up to 80%, resulting in
crop failure if the pest is not controlled (Marwoto and Suharsono, 2008).
Efforts to control pests can be done with mechanical/physical, biological or chemical
methods. The control of this pest has so far, been done mostly by the use of chemical method
through application of synthetic insecticides, but this method results in a negative impact on
insects, environment and human health. The negative impact on insects is the occurrence of
pest resistance to synthetic insecticides, the pest resurgence or the killing of natural enemies
of pests. The negative impact on the environment can be the accumulation of synthetic
insecticide residue on the farm and in the crops harvested. The negative impact on humans
coulb be an accident on the users of synthetic insecticides and various diseases in humans
such as skin irritation, disruption of the nervous system and cancer likelihood as a result of
intake of food contaminated with synthetic insecticides (Rogers, 2010).
Pesticide residue is pesticide remains on farmland and in agricultural yield that are not
decomposed. Some studies indicated that there are pesticide residues on land and agricultural
production. Karyadi, et al., (2011) observed an increase in heavy metals levels (Pb) on onion
crop land in Kendal, Centre Java province due to the use of pesticides. This happened
because the farmers in Kendal used seven kinds of pesticides containing heavy metal Pb. Pb
levels increased by 43 071.60 mg/Ha compared to the land condition before planting and
after onions harvest. Spraying frequency, the pesticides dose, and variable content of Pb in
pesticides significantly affected the content of Pb in the soil. One season of onion planting
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can increase the Pb content in the soil as much as 2 991.26 mg/Ha. Pesticide residues have
also been found on the agricultural yields. Mutiatikum and Sukmayati (2009) found
insecticide residues on the samples of local and imported rice from the cities of Cianjur,
Semarang and Surabaya. In Surabaya and Semarang, residues of insecticides (lindane, aldrin
and heptaklor) were found on rice in low levels. Although there were only small amounts of
residues, it should be a cause for concern because they have a long half-life. Similar results
were also found in various vegetables, like red peppers, lettuce, and onions (Miskiyah and
Munarso, 2009); spinach (Amarantus indica), water spinach (Ipomoea aquatica), mustard
(Brassica juncea) and beans (Vigna sinensis) (Tuhumury, et al., 2012), as well as in carrots
(Daucus carota) (Ohorella, et al., 2013). Pesticide residues were also found in red peppers,
lettuce, and onion which were obtained from farmers, traders, and markets in vegetable
production centers at Central Java and West Java (Miskiyah and Munarso, 2009).
Because of the negative impact of synthetic insecticides applied as the most common way to
control pests on plants, there is need for efforts towards the control of pests with methods that
are more friendly to the environment and human health. This can be effective by the use of
botanical insecticides which are biodegradable and safe for the environment and human
health. Botanical insecticides do not kill insect pests directly, but they can reduce the
incidence of pest attacks by acting as insect repellents or interfering with the development of
eggs, larvae and pupae of insects; inhibit skin turnover insects; disturb insect communication;
inhibit the ability of eating insects provisionally or permanently (as antifeedant); inhibit the
reproduction of insects or can attract insects (as attractant) (Ware, 1983).
Bitter gourd is one of the plants in Cucurbitaceae family which has been widely studied as a
botanical insecticide. The parts of its plant which have been studied are the leaves (Yasui,
2002; Ling, et al., 2008; Devanand and Rani 2008; and Abe and Matsuda, 2000). Bitter gourd
fruit has also been studied by Singh, et al., 2006 and Maurya, et al., 2009, but the age of its
fruit which is most effective as botanical insecticide has not been studied.
Wild bitter gourd (Momordica charantia L.) is an annual plant that can grow well at an
altitude 0-1000 m asl. Wild bitter gourd plant morphology is similar to the characteristics of
bitter gourd cultivated for food purposes, but it has special characteristic. The wild one has
small rash resembling a thorn in the entire skin surface of the fruit. The other characterictic is
that the size of the fruit is much smaller than the green bitter gourd, which is around the size
of a thumb about 2-5 cm in fruit length.
Wild bitter gourd plants are vines or climbing, it has tendrils strong stems and smelled
dreadful. The stems can reach 2 to 3 m. The plant roots is the taproot type. The leaves
arranged alternate, the leaves width can reach 10 cm. Pare leaves are oval and have fur on
their surface. Leaf blade pare split almost to the leaf base. The leaves are shaped like a finger
bone. The plant is monoceous. The flowers are yellow and grow from the armpit. Flowers
began to appear around the age of 45-55 days after sowing. There are male (staminate) and
female flowers (pistilate) on the same plant. The female one become fruit after pollination.
The fruit is elliptical. Unripe fruit is green. The ripe one turn to orange or orange yellowish
and brake into three pieces. The seeds is light brown to black. It is flat round shape and
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jagged edge of a flat section. The seeds are about 5-9 mm length and 2.5 to 6 mm width
(Holm, et al., 1997).
Bitter gourd has function as a botanical insecticide. Yasui (2002) showed that the methanol
extract of bitter gourd leaves effectively hinder eating larvae of Spodoptera litura and
Pseudolatia separata. Ling, et al. (2008) stated that in the extract of bitter gourd leaf there
are triterpenoids compounds which are effectivelly hinder the eating of Plutella xylostella
larvae.
The ethanol extract of bitter gourd leaves inhibited the oviposition of Liriomyza trifolii
imago on bean leaves treated with the extract. The concentrations of 2000-4000 mg/ml
ethanol extract of bitter gourd leaves showed antifeedant and antioviposisi activity in L.
sativae Blanchard imago (Diptera: agromyzidae) significantly. Antifeedant index (AFI) of
cyclohexane extract of bitter gourd leaves with a concentration of 1000 ug/ml for 2 days in
imago L. sativae was 11.08%. While at the same concentration, AFI of ethyl acetate extract
and n-butanol extract of bitter gourd leaves at the same concentration was 34, 89%, and
22.99%, respectively. The AFI lowest value was water extract that was 0%. Ethyl acetate
extract of the bitter gourd leaves had the highest bioactivity (Ling, et al., 2009).
Acetone extract of bitter gourd leaves showed toxic activity and greatly impede the ability
feeding of the Spodoptera litura Fab. larvae instar 3. Tests was conducted by feeding that has
been treated acetone extract of bitter gourd leaves with a dose of 100 mg / 21cm2. LD50 value
of the acetone extracts of bitter gourd leaves was 72.60 mg/21 cm² (Devanand and Rani,
2008). Meanwhile, the methanol extract of bitter gourd leaves hindered eating four species of
Cucurbitaceae family beetles, namely the Aulacophora femoralis, A. nigripennis, Epilachna
admirabilis and E. boisduvali (Abe and Matsuda, 2000).
Besides the leaves, bitter gourd fruit also has a botanical pesticide potency. Bitter gourd fruit
juice and hexane extract of bitter gourd fruit serves as a larvicide in mosquito larvae of
Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti (Singh, et al., 2006), likewise
with petroleum ether extract, carbon tetrachloride extract and methanol extract of bitter gourd
fruit. Those extracts were also capable of controlling larvae of A. stephensi and C.
quinquefasciatus (Maurya, et al., 2009). Petroleum ether extract of bitter gourd fruit was
more effective than the karbontertraklorida extract in controlling larvae of A. stephensi and C.
quinquefasciatus. Petroleum ether extract has LC50 lower than the carbon tetrachloride extract.
LC50 petroleum ether extract of bitter gourd fruit on the larvae of Anopheles stephensi was
27.60 ppm (treatment for 24 hours) and 17.22 ppm (treatment for 48 hours), and LC90
petroleum ether extract of bitter gourd fruit on the larvae of Anopheles stephensi was 154.99
ppm and 94.79 ppm respectively for treatment 24 hours and 48 hours. While LC50 of carbon
tetrachloride extract of bitter gourd fruit for larvae of A. stephensi was 49.58 ppm (treatment
for 24 hours) and 16.15 ppm (treatment for 48 hours), and LC90 for the treatment of 24 and 48
hours also was 521.02 ppm and 369.99 ppm (Maurya, et al., 2009).
Study results showed that botanical insecticide active compounds that had been isolated from
the leaves extract of bitter gourd was terpenoids (Mekuria, et al., 2005), (Yasui, 2002), (Ling,
et al., 2008), (Abe and Matsuda, 2000) and (Devanand and Rani, 2008). Terpenoids in plants
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was part of plant self-defense mechanism against pests and diseases, so it was used much as
insecticides, fungicides and herbicides in agriculture, and as antimicrobial and antifungal in
pharmaceutical field. Terpenoids provide benefits for the plant by refusing (repellent) or
killing predators or disease. However, terpenoids can also be counterproductive for plants
because it can act as attractants. The effectiveness of terpenoids for the plant defense was
affected by the total number of existing terpenoids, the number of existing specific terpenoids
and was also influenced by the composition of two or more existing terpenoids.
Chrysothamnus nauseosus plants during the summer was protected from pests due to the high
content of terpenoids, which was about 80 ug/g dry weight. Terpenoids contained by the plant
consists of α- and γ-muurolen, β-humulen and E-β-farnesen. In winter, the content of these
terpenoids declined so that deer can eat these plants (Tellez, et al., 2002).
Yasui (2002) found two kinds of active compounds obtained via silica gel chromatography
purified by HPLC in bitter gourd, namely monoglukosida triterpenoids (momordisin II) and
diglukosida triterpenoids (momordisin I). Triterpenoids identified as
3,7,23-trihidroksicucurbita-5,24-dien-19-al (momordisin I) which were isolated from the
leaves of bitter gourd inhibit oviposition of Liriomyza trifolii on host plant leaves treated
with 33.60 g/cm2 leaf surface (Mekuria, et al., 2005). Ling, et al., 2008 also found that
momordisin I and momordicine II were also active in inhibiting eating ability of Plutella
xylostella larvae instar 2 and 3. Momordisin II showed antifeedant effect significantly on P.
separata on artificial food with concentrations of 0, 02; 0.1 and 0.5% momordisin II (Yasui,
2002). Each momordisin I and II did not affect the eating ability of Epilachna admirabilis
and E. boisduvali, but the eating ability was inhibited by mixture of momordisin I and II or II
momordisin mixtures with other components (Abe and Matsuda, 2000).
Armyworm (Spodoptera litura Fab.) was a polyfaghous pest which was widespread in Asia
and Africa. It was classified into insect which undergo perfect stage of metamorphosis,
namely egg-larva-pupa-adult insect (imago) stage. The larvae hid during the day and searched
for food at night. The eggs laid by the female imago in groups on the underside of the leaf
surface at night. It was covered by an orange brown like cotton layer. The eggs laid by the
female imago average can reach until 400 eggs in 3-4 groups each time laying. Each group
totaled 80-150 eggs, so total eggs laid were up to 1 500-2 500 eggs within 6-8 days. The eggs
were round, slightly flattened with a diameter of 0.4-0.7 mm. The incubation eggs period
lasted in 3-5 days. The larvae newly hatched had measuring 2.00 to 2.74 mm (Alyokhin, et al.
2012).
Larvae grew through 5-6 instar periods. At instar 1-3 it remained on the underside of leaves
surface. At instar 4-6, it will dropped to the ground, loosened the ground, and prepared the
clay for the cocoon. The shape of last period instar was fat and smooth with a length of about
40-50 mm. The larva period lasted about 20-28 days (Sullivan, 2007).
Color of the pupa was maroon and it lied in the ground. The length was about 18-22 mm. The
last segment of the stomach was shaped like two hooks. Pupal period lasted 7-11 days. The
imago had a yellowish-white body. The front wings were dark brown with light shadow lines
and stripes. Hind wings were white with violet sheen satin and brown stripe. The head was
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like having light and dark brown tufts. The body length ranged from 14-18 mm. Wing span
reached 28-38 mm (Sullivan, 2007). According to Alyokhin, et al. (2012) the armyworm life
cycle took place within 30-40 days.
The wild bitter gourd had not been studied as botanical insecticide until now, either. So, this
study used water solvent to extract the wild bitter gourd fruit because it was universal solvent
which easily to find, economical and friendly to environment.
2. Research Methods
2.1 Materials and Tools
The research was conducted at Biology Laboratory, Agriculture Faculty, Widyagama Malang
University, Indonesia for toxicity test of water extract of wild bitter fruit on armyworm larvae
instar 2. This was also conducted at Chemical Laboratory of Polytechnic Malang for active
compound test. Materials used in this study were three levels of fruit maturity of wild bitter
gourd, and armyworm larvae instar two. The tools used in this study were grinder, sifter 60
mesh, centrifuge, waterbath, plastic cups, glassware, digital scales and LC-MS/MS (Liquid
Chromatography-Tandem Mass Spectrometry).
This research consisted of two studies: first was the test using HPLC-MS/MS to determine
the chemical compounds in the water extract and second was the toxicity test of the extract on
armyworm as antifeedant and mortality tests. HPLC-MS/MS test was conducted according to
the determination procedure conducted by Ma, et al., 2012.
2.2 Water Extracts of Wild Bitter Gourd Fruit Making
Wild bitter gourd fruits on this research were from Tawang Argo Village, Karangploso
subdistrict, Malang, Indonesia. The village had a hilly topography with a height of 700 m asl.
Flower of wild bitter gourd was observed first to know the life span from anthesis to riping
fruit. Based on data which was 14-15 days, then the three maturity levels fruit of wild bitter
gourd were determined:
- 4H: fruit maturity level 4 days after anthesis;
- 8H: fruit maturity level 8 days after anthesis;
- 12H: fruit maturity level 12 days after anthesis.
The female flower of wild bitter gourd was marked with a label. The fruit was harvested in
accordance with the age of the treatment which was 4, 8 and 12 days after anthesis, then was
sliced thinly and dried in the winnowing and covered with black cloth on the shady
conditions for 5 days. The harvest of fruit maturity level 4H was as many as 770 pieces
weighing 271.20 grams, while fruit maturity levels 8H and 12H were as many as 652 pieces
weighing 1 143.93 grams, and 727 pieces weighing 2 773.44 grams. After that, the fresh wild
bitter gourd fruit was dried for 5 days, blended into powder and sieved with 16 mesh sieve.
The powder obtained from the fruit maturity levels 4H, 8H and 12H respectively were as
much as 31.19 grams, 30.64 grams, and 30.03 grams dry weight, while the water contents on
the powder at fruit maturity levels 4H, 8H and 12H were 9.97%, 10.1% and 10.3%,
respectively.
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The powder of wild bitter gourd fruit maturity levels 4H, 8H and 12H was extracted by
maceration with water solvent based on the procedure performed by Balafif, et al. (2013).
The powder as much as 30 grams was extracted by maceration with distilled water for 3 x 24
hours. On day 1 the powder was macerated with 180 ml of distilled water, as well as day 2
and day 3. Total distilled water was added to 540 ml. The filtrate result from day 1, day 2 and
day 3 was centrifuged twice each at a speed of 3 000 rpm for 5 minutes. The filtrates obtained
from treatments 4H, 8H and 12H were respectively 117.5 ml, 115.0 ml and 116.3 ml. The
filtrates subsequently were thickened with a water bath at a temperature of 100° C up to
one-third volume, which was to be 38.3 to 39.2 ml.
2.3 The Phytochemical Test
Water extract of wild bitter gourd fruit contained saponin if there was stable foam when it
was given hot water and shaking vigorously after cooling. The extract contained alkaloids if
there was white sediment after water extract sample was treated with Mayer and there was
brown sediment after extract sample was reacted with Dragendorf reagent. Water extract
contained any steroids if appeared green or blue color after the sample was reacted with the
Liebermann-Burchard reagent. This was indicated by the onset of a yellow color after the
sample reacted with 2 N H2SO4 solution, the incidence of red color after the sample was
treated with NaOH 10% solution and the incidence of yellow color accompanied with foam
after the sample water extract was treated with concentrated HCl concentrated, and
Magnesium. The water extract contained phenolic compounds indicated. by the appearance of
a black color after the sample was treated with 5% FeCl3 solution. Water extract contained
triterpenoids if appeared brownish red color after the sample was reacted with the
Liebermann-Burchard reagent.
2.4 The Bioactive Compounds Test with LC-MS/MS
Operating Conditions of Liquid Chromatography-Tandem Mass Spectrometry were as
follows. Column used had as specifications Hypersil Gold (50mm x 2.1 mm x 1.9 m).
UHPLC brands ThermoScientific 1250 ACCELLA type which consisted of vacuum degasser,
quartener pump, an autosampler thermostatically controlled with a personal computer
through a program called x-calibur 2.1. A mobile phase consisted of 0.1% formic acid in
aquabidest, phase B consisted of 0.1% formic acid in Acetonitrile, phase C consisted of 0.1%
formic acid in methanol. A linear gradient at a rate of 300 mL/minute with a mobile phase
was set as follows: a) 0-0.6 (minute 65% A, 25% B, 10% C), 2-3.5 minute 90% B, 10% C),
4-5 minutes which was equal to 0-0.6 minutes. Injection volume on LC was 10 µL . The
column was controlled at 30°C, and autosampler compartment was set at 10°C. The usage of
MS/MS Triple Q (quadrupole) mass spectrometer TSQ Quantum ACCESS MAX from
Thermo Finnigan with ionization source ESI (electro spray ionization) was controlled with
software TSQ Tune-operated with positive mode.
2.5 Antifeedant Test on Armyworm Larvae
The aim of this test was to determine the level of fruit maturity and concentration of water
extract of wild bitter gourd fruit which was effective as antifeedant test on each level of fruit
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maturity. The tests were carried out with five concentrations of water extract of wild bitter
gourd fruit and control. Each treatment concentrations of water extract and controls was
repeated 4 times using a randomized block design. Pest target was 20 armyworm larvae instar
two in each treatment unit. The concentrations of water extracts used were 20%, 30%, 40%,
50% and 60%. To make 20%, 30%, 40%, 50% and 60% concentration, 2, 3, 4, 5 and 6 ml
concentrated solution of water extract were taken and put in a 10 ml flask and then was added
distilled water until the solution volume reached 10 ml.
The treatment was conducted by dipping larvae feed into each extract water according to
maturity level and treatment concentration. Feed given to the armyworm larvae was cabbage
which is the common feed used in mass breeding of armyworm. Cabbage was purchased
from traditional markets and was washed first with water and then dried. The cabbage was
cut circular with diameter of 3 cm. Pieces of cabbage were then dipped in water extract and
air-dried for five minutes. They were put into a plastic cup with 9 cm diameter on the top side,
diameter 7 cm on below side and height of 7 cm. For the control treatment, the pieces of
cabbage were dipped in distilled water only. Into each plastic cup was inserted one hungry
armyworm larvae. Filter paper soaked in 0.5 ml water was placed on the base of plastic cups
to maintain moisture. The treatments were conducted over 24 hours.
The tests were carried out with non-choice method (Pavela, 2009). Observations were carried
out 24 hours after treatment. The rest of leaves that were not eaten by the armyworm larvae
was calculated to obtain leaf area eaten. Percentage antifeedant index (AF) was calculated
using the formula:
AF (%) = (C - T) / (C + T) x 100% (Pavela, 2009),
where C: leaf area eaten by larvae on control, T : leaf area eaten by larvae on treatment.
Water extract which had the greatest value of antifeedant index was the most active extracts
for antifeedant test.
2.6 Mortality Test
The aim of this test was to determine the level of maturity of bitter gourd fruit and
concentration of water extract which was effective as a mortality test. The method used was
the same as the antifeedant test method. Observations were made every 24 hours after
treatment up to 6 x 24 hours, ie at 24, 48, 72, 96, 120, 144 hours after the treatment. Number
of armyworm larvae that died in every 24 hours and the percentage mortality was observed.
Percentage armyworm larvae that died was then calculated based on the following formula:
(X-Y) / X x 100% (Abbott, 1987),
X: live armyworm larvae on control
Y: live armyworm larvae on the treatments
2.7 Data Analysis
The data on antifeedant and mortality test was analyzed by F test at error level α = 5%, and if
there were significant results, the analysis was continued with HSD test (Steel and Torrie,
1960).
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3. Result and Discussion
3.1 Phytochemicals test of Water Extract of Wild Bitter Fruit
The phytochemical test result are stated in Table 1.
Table 1. Secondary metabolites in water extract of wild bitter gourd fruit
No.
Secondary Metabolite
Reagent
Result
1.
Saponin
Hot water
-
2.
Alkaloid
Mayer reagent
-
Dragendorf reagent
-
3.
Steroid
Liebermann-Burchard reagent
-
4.
Phenolic
FeCl3 5%
+
5.
Flavonoid
H2SO4 2N
+
NaOH 10%
+
Concentrated HCl + Mg
+
6.
Triterpenoid
Liebermann-Burchard reagent
+
This was consistent with the research conducted by Horax, et al. (2005) which states that
there was phenolic compounds in the extract of bitter gourd fruit. Horax, et al. examined the
total phenolic content and acid phenolic components in 4 bitter gourd varieties which were
India White, India Green, China White and China Green. Total phenolic content obtained
from samples dried with oven was much higher than from samples that went through freeze
drying. Total phenolic content obtained from bitter gourd fruit pulp extract was 5.36 to 8.90
mg CAE/g dry weight. Total phenolic content of the bitter gourd seeds extract was lower than
that contained in the bitter gourd fruit which was 4.67 to 8.02 mg CAE/g dry weight. Major
phenolic acids found were gallic acid, gentisic acid, catechin, epicatechin and chlorogenic
acid. Meanwhile, protocatecheuic acid, sirinic acid and benzoic acid were also found in the
extract but in small amounts, ie less than 10 mg/100 g material dry weight. Similar results
were also demonstrated by Ghaima, et al. (2013) who found phenolic compounds in bitter
gourd fruit extract.
Water extract of wild bitter gourd fruit contained flavonoids. This was in line with the
observation by Tan, et al., 2014, that the bitter gourd fruit extract contains flavonoid
compounds. Total flavonoid contained in the water extract was very small, ie only 5.4%
compared with the flavonoids content presented in the acetone extracts. Mada et al. (2012)
suggested that flavonoids were also found in the water extract of bitter gourd leaves.
Wild bitter gourd fruit water extract contained triterpenoids. This was in accordance with
Sundari, Padmawati and Ruslan (1996) who reported that bitter gourd flesh contained
steroid/triterpenoid. Nagarani, et al. (2014) also stated that other parts of bitter gourd plants
contained cucurbitane triterpenoids, phenolic compounds, glucoside and several types of
peptides. However, Nagarani, et al. (2014) regretted that there was still very little information
about bioactive compounds in wild bitter gourd.
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3.2 Bioactive Compounds in Water Extract of Wild Bitter Gourd Fruit
Preparation and procedures for testing the active compound content was like that
conducted by Ma, et al., 2012. The results of analysis of the active compounds in water
extract of wild bitter gourd fruit maturity levels 4H, 8H and 12H are presented in Table 2.
Table 2 shows that there were 4 kinds of triterpenoid compounds in water extract of
bitter gourd fruit maturity level 4H, 8H and 12H, namely momordicoside L,
momordicoside K, 3β, β7, 25-trihydroxycucurbita-5.23(E)-diena-19-al: R1=H, R2=H and
momordicine 1. The structure of those compounds are shown in Figure 1.
Figure 1. The Structure of Momordicoside L, Momordicoside K, 3β, β7,
25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2=H and momordicine 1
(Ma, et al., 2012).
Tabel 2. Active Compounds and the Area Found in the Water Extract of Wild Bitter Gourd
Fruit Maturity Levels 4H, 8H and 12H
Fruit Maturity
Level
Active Compound
H
Momordicoside L
Momordicoside K
3β,7β,25-trihydroxycucurbita-
5,23(E)-diena-19-al:R1=H,R2=H and Momordicine 1
8H
Momordicoside L
Momordicoside K
3β,7β 25-trihydroxycucurbita-5,
23(E)-diena-19-al:R1=H,R2=H and Momordicine 1
12H
Momordicoside L
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Momordicoside K
3β,7β, 25-trihydroxycucurbita-
5,23(E)-diena-19-al:R1=H,R2=H and momordicine 1
The compounds area found in the water extract of fruit maturity level 4H was the widest, so it
can be stated that active compound in the water extract of fruit maturity level 4H was the
highest. In water extract of fruit maturity level 4H the content of momordicoside L was 2.73
and 3.99 higher than those contained in the water extract of fruit maturity levels 8H and 12H.
Momordicoside L content in water extracts of fruit maturity level 8H were 1.46 times higher
than those contained in water extract of fruit maturity level 12H. Meanwhile, momordicoside
K content in water extracts of fruit maturity level 4H was also the highest when compared to
the content of water extracts of fruit maturity levels 8H and 12H. The amount of
momordicoside K in water extract of fruit maturity level 4H was respectively 16.86 and 2.81
times higher than that contained in the water extract of the fruit maturity levels 8H and 12H.
While momordicoside K contained in the water extract of fruit maturity level 8H was less
than that contained in water extract of fruit maturity level 12H, ie 0.17 times. The amount of
compound 3β, β7, 25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2=H and
momordicine 1 at fruit maturity level 4H was respectively 23 times and 103 times higher than
at fruit maturity level 8H and 12H. These results are in line with that expressed by the
Maharani (2013) that different age affected content and types of constituent compounds in the
different plant parts.
Momordicoside L in water extracts of fruit maturity level 4H, 8H and 12H can be seen on
Figure 2-7.
Figure 2. Momordicoside L in Water Extract of Fruit Maturity Level 4H of Wild Bitter Gourd
Figure 3. Area of Momordicoside L in Water Extract of Fruit Maturity Level 4H of Wild
Bitter Gourd
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Figure 4. Momordicoside L in Water Extract of Fruit Maturity Level 8H of Wild Bitter Gourd
Figure 5. Area of Momordicoside L in Water Extract of Fruit Maturity Level 8H of Wild
Bitter Gourd
Figure 6. Momordicoside L in Water Extract of Fruit Maturity Level 12H of Wild Bitter
Gourd
Figure 7. Area of Momordicoside L in Water Extract of Fruit Maturity Level 12H of Wild
Bitter Gourd
The comparisan of momordicoside L compound which found in water extracts with fruit
maturity levels 4H, 8H and 12H. The figures can be seen on Figure 8-13.
Figure 8. Momordicoside K in Water Extract of Fruit Maturity Level 4H of Wild Bitter Gourd
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Figure 9. Area of Momordicoside K in Water Extract of Fruit Maturity Level 4H of Wild
Bitter Gourd
Figure 10. Momordicoside K in Water Extract of Fruit Maturity Level 8H of Wild Bitter
Gourd
Figure 11. Area of Momordicoside K in Water Extract of Fruit Maturity Level 8H of Wild
Bitter Gourd
Figure 12. Momordicoside K in Water Extract of Fruit Maturity Level 12H of Wild Bitter
Gourd
Figure 13. Area of Momordicoside K in Water Extract of Fruit Maturity Level 12H of Wild
Bitter Gourd
Comparison of compound 3β, β7, 25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2=H
and Momordicine 1 among the water extracts with fruit maturity levels 4H, 8H and 12H
(Figure 14 -19).
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Figure 14. 3β, 7β, 25-trihydroxycucurbita-5,23(E)-diena-19-al:R1=H, R2=H and
Momordicine 1 in Water Extract of Fruit Maturity Level 4H of Wild Bitter Gourd
Figure 15. Area of 3β, 7β, 25-trihydroxycucurbita-5,23(E)-diena-19-al:R1=H, R2=H and
Momordicine 1 in Water Extract of Fruit Maturity Level 4H of Wild Bitter Gourd
Figure 16. 3β, 7β, 25-trihydroxycucurbita-5,23(E)-diena-19-al:R1=H, R2=H and
Momordicine 1 in Water Extract of Fruit Maturity Level 8H of Wild Bitter Gourd
Figure 17. Area of 3β, 7β, 25-trihydroxycucurbita-5,23(E)-diena-19-al:R1=H, R2=H and
Momordicine 1 in Water Extract of Fruit Maturity Level 8H of Wild Bitter Gourd
Figure 18. 3β, 7β, 25-trihydroxycucurbita-5,23(E)-diena-19-al:R1=H, R2=H and
Momordicine 1 in Water Extract of Fruit Maturity Level 12H of Wild Bitter Gourd
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Figure 19. Area of 3β, 7β, 25-trihydroxycucurbita-5,23(E)-diena-19-al:R1=H, R2=H and
Momordicine 1 in Water Extract of Fruit Maturity Level 12H of Wild Bitter Gourd
3.4 Antifeedant Test
Based on the analysis of variance, the interaction between fruit maturity levels and the
concentrations of water extract affected antifeedant index significantly, so the anylysis was
continued with honestly significant difference to find the different treatments. Analysis of
variance was based on arcsin transformation of antifeedant index data. The antifeedant index
can be seen on Table 3.
Table 3. Interaction Between Fruit Maturity Levels and Concentrations of Water Extract of
Wild Bitter Gourd Fruit on Antifeedant Index (%)
Treatments
Antifeedant Index (%)
4H 20%
28,10 cd
4H 30%
29,96 de
4H 40%
34,22 f
4H 50%
40,08 g
4H 60%
44,20 h
8H 20%
26,35 cd
8H 30%
28,52 cd
8H 40%
28,70 de
8H 50%
32,62 ef
8H 60%
33,05 ef
12H 20%
20,75 a
12H 30%
22,03 ab
12H 40%
26,29 cd
12H 50%
25,09 bc
12H 60%
28,22 d
Note: Numbers followed by the same letter on the same column were not significantly
different by HSD test with α = 5%
At fruit maturity level 4H of wild bitter gourd, concentrations 20% and 30% did not differ on
the antifeedant index. Concentrations 20% and 30% caused the lowest antifeedant index,
when compared to the concentrations 40%, 50% and 60%. This showed that the water extract
of bitter gourd fruit at a concentration 20% and 30%, had the lowest ability to inhibit eating
ability of armyworm larvae instar two. At fruit maturity level 4H of bitter gourd, water
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extract concentration 60% gave the highest antifeedant index value, which meant that this
concentration had the highest inhibition on eating ability of armyworm larvae.
At fruit maturity level 8H of wild bitter gourd, a similar phenomenon occured with the fruit
maturity level 4H of wild bitter gourd. The antifeedant index values at concentration 20% and
30% were not different. Both concentrations led to the same inhibition of the eating ability of
armyworms larvae instar two. Antifeedant index values of the concentration 50% and 60%
did not differ with the antifeedant index at concentration 40%, either, but they were higher
than the antifeedant index at concentration 20% and 30%. Concentration 50% and 60% of
water extract of fruit maturity level 8H were able to inhibit eating ability of armyworm larvae
instar two greater than concentration 20% and 30%. The inhibition of eating ability of
armyworm larvae instar two at concentration 50% and 60% were 32.62% and 33.05%.
In the water extract of fruit maturity level 12H, concentration 20% and 30% also had lower
antifeedant index value than concentrations 40% and 60%. Antifeedant index on
concentration 40% was not different with concentration 50% and 60%, but different with
concentration 20% and 30%. The highest antifeedant index was at the concentration 40% and
60%.
Comparing among the fruit maturity levels 4H, 8H and 12H, antifeedant index at
concentrations 20% and 30% at fruit maturity level 4H was not different with the antifeedant
index at fruit maturity level 8H, but in contrast with fruit maturity level 12H. Antifeedant
index on extracts of fruit maturity level 4H was higher than on extracts of fruit maturity level
12H. Antifeedant index on extracts from fruit maturity level 8H was also higher than the
antifeedant index of the extract with the fruit maturity level 12H.
Among high concentrations, ie 50% and 60%, it was found that the water extract of 4H
maturity level of wild bitter gourd fruit had higher antifeedant index value than fruit maturity
levels 8H and 12H. Antifeedant index of water extract of maturity level 8H of wild bitter
gourd fruit was also higher than fruit maturity level 12H treatment.
This occured because the water extract of younger wild bitter gourd fruit, i.e. in fruit maturity
level 4H had more content of triterpenoid compounds than in fruit maturity level 8H and 12H.
Based on the chemical compounds analysis, in the fruit maturity level 4H, 8H and 12H,
triterpenoid compounds found were momordicoside K, momordicoside L, compound 3β, β7,
25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2=H and Momordicine 1. In water
extract of fruit maturity level 8H and 12H there were also found such triterpenoid compounds
but in lower contents than in fruit maturity level 4H. Momordicoside L content in fruit
maturity level 4H was 2.73 and 3.99 times higher than that found in 8H and 12H fruit
maturity level. Momordicoside K content in the water extract of 4H fruit maturity level 4H
was also the highest. The compound was 16.86 and 2.81 times higher than that contained in
fruit maturity levels 8H and 12H. While 3β, β7,
25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2=H and Momordicine 1 found in water
extract of fruit maturity level 4H was 23,20 and 103,0 and times higher than in fruit maturity
level 8H and 12H.
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This was in line with Dewi (2013) who said that different ages of fruit had different contents
of secondary metabolite compounds. The research result on 3 levels of fruit maturity
indicated that the content of secondary metabolites in fruits with different ages were different.
Dewi (2013) working on the bel fruit (Limonia acidissima) showed that the methanol extract
of ripe bel fruit had higher bioactivity than the methanol extract of old and young bel fruit.
The secondary metabolite content in bel fruit, ie flavonoids, increased with the incerasing age
of bel fruit. Rahardjo, Darwati and Shusena (2006) demonstrated similar results as well, that
the organs of plants at different ages produced secondary metabolites with different contents.
Secondary metabolites contained in canopy of purwoceng (Pimpinella pruatjan Molkenb) at
nine months of age was higher than for the age of three and six month. Tellez et al. (2002)
stated that the effectiveness of secondary metabolites for plant defense was affected by the
total number of existing compounds, and the content and composition of the compounds.
3.5 Mortality Test
Analysis of variance test results indicated that the interaction between the treatment of fruit
maturity level of wild bitter gourd and concentration of the water extract did not significantly
affect the percentage of armyworm larvae mortality. In the fruit maturity levels 4H, 8H and
12H of wild bitter gourd, on the observations of day 1 to day 6, the concentration of water
extracts did not significantly affect the percentage of armyworm larvae mortality. On the
observations on day 7, extract concentration on the fruit maturity level 4H significantly
affected the percentage of armyworm larvae mortality, while the extract concentration on the
fruit maturity levels 8H and 12H were not significant. Mortalities of armyworm larvae can be
seen on Table 4.
Table 4. Armyworm Larvae Mortality at Water Extract Concentrations of Fruit Maturity
Level 4H (%)
Concentration
Observation Day-
1
2
3
4
5
6
7
20%
0.00
1.25
1.25
5.00
8.75
25.00
30.26a
30%
0.00
0.00
0.00
2.50
7.50
18.42
35.53a
40%
0.00
3.75
5.00
11.25
13.75
28.95
44.74ab
50%
0.00
1.25
2.50
5.00
10.00
42.11
59.21ab
60%
0.00
2.50
5.00
10.00
11.25
47.37
68.42b
The numbers in the same column followed by the same letter do not differ by HSD test with
level α = 5%
Fruit maturity level 4H on the observation day 7 showed that armyworm larvae mortality at
concentrations 20% and 30% were lower than at 60% concentration treatment. Meanwhile,
the armyworm larvae mortality at concentration of 40% treatment did not differ with
concentrations 50% and 60%.
At fruit maturity level 8H observation day 7, armyworm larvae mortality at concentration
20%, 30%, 40%, 50% and 60, respectively, was 27,63%, 34,21%, 40,79%, 47,37%, and
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48,68%. While at fruit maturity level 12H observation day 7, armyworm larvae mortality at
concentration 20%, 30%, 40%, 50% and 60, respectively, was 26,32%, 27,63%, 28,95%,
31,58%, and 36,84%.
Based on probit analysis to determine LC50 values, at fruit maturity level 4H observations day
7, probit model obtained was as follows:
Y = 0.844 + 2.121 x,
where x = concentration based on log10 transformed. At the fruit maturity level 4H, the
estimated LC50 was the concentration 40%.
4. Conclusion
Water extract of wild bitter gourd fruit contained phenolic, flavonoids and triterpenoids, but
did not contain saponin, alkaloids, and steroids.
The active compounds contained in the water extract of wild bitter gourd fruit at maturity
level 4H, 8H and 12H are momordicoside L, momordicoside K, compound 3β, β7,
25-trihydroxycucurbita-5.23(E)-diena-19-al:R1=H, R2= H and momordicine 1.
Interaction between fruit maturity levels 4H, 8H and 12H and the concentration of the water
extract resulted in a significantly different of antifidant index at non-choice method. Fruit
with a maturity level 4H 50% and 60% resulted the highest antifeedant index, ie 40.08% and
44.20%.
In mortality test, there was no interaction between fruit maturity level and extract
concentration. Fruit maturity level gave no significant result on mortality armyworm larvae,
except on observation day 7 at fruit maturity level 4H. LC50 at fruit maturity level 4H on
observation day 7 was 40%.
Acknowledgement
The research was financed by Kopertis VII East Java and Directorate General of Higher
Education, the Ministry of Education and Culture of Indonesia.
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Momordica species (Family Cucurbitaceae) are cultivated throughout the world for their edible fruits, leaves, shoots and seeds. Among the species of the genus Momordica, there are three selected species that are used as vegetable, and for medicinal purposes, Momordica charantia L (Bitter melon), Momordica foetida Schumach (Bitter cucumber) and Momordica balsamina L (African pumpkin). The fruits and leaves of these Momordica species are rich in primary and secondary metabolites such as proteins, fibers, minerals (calcium, iron, magnesium, zinc), β-carotene, foliate, ascorbic acid, among others. The extracts from Momordica species are used for the treatment of a variety of diseases and ailments in traditional medicine. Momordica species extracts are reputed to possess anti-diabetic, anti-microbial, anthelmintic bioactivity, abortifacient, anti-bacterial, anti-viral, and play chemo-preventive functions. In this review we summarize the biochemical, nutritional, and medicinal values of three Momordica species (M. charantia, M. foetida and M. balsamina) as promising and innovative sources of natural bioactive compounds for future pharmaceutical usage.
... It has been used in ethnomedicine and has been proposed as a source of anti-diabetic drugs and antioxidants (Virdi et al., 2003;Grover and Yadav, 2004;Kubola and Siriamornpun, 2008;Joseph and Jini, 2013;Desai and Tatke, 2015). It has triterpenes, terpenoids, phenolics and other compounds that act as antifeedants to insects, and the antifeedant or repellent activity has been tested against some lepidopteran pests (Yasui, 2002;Ling et al., 2008;Wardhani et al., 2015) and occasionally against coleopteran and dipteran pests (Chandravadana and Pal, 1983;Abe and Matsuda, 2000;Mekuria et al., 2005). Thus it has a good potential of acting as an antifeedant that can repel insect pests and at the very least be harmless to human consumers. ...
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Bitter gourd (Momordica charantia L.) has compounds that repel insect pests. Unlike conventional pesticides, these compounds are eco-friendly and beneficial for human health. However the mechanisms by which these compounds repel insects and affect their physiology remains poorly known. Here we used Drosophila melanogaster (Meigen) to address these issues. We tested a wild strain, and a laboratory bred Canton S strain. Bitter gourd extract reduced the viability of developing flies, but did not affect survival in adults. Flies avoided bitter gourd extract in a food choice assay, and consumed a significantly low amount of food mixed with bitter gourd – indicating that it acts as an antifeedant. Transgenic flies with impaired aversive taste sensitive neurons showed a reduced aversion towards bitter gourd extract showing that these compounds act through bitter sensitive gustatory neurons. Finally, flies also retained the memory of consuming bitter gourd extract for at least 24 hours, suggesting an additional cognitive mechanism for long term aversion. Our study provides the first evidence of bitter gourd compounds acting as antifeedants and also as potent reinforcers of aversive memory in drosophilids. We suggest that flies can be used to understand the physiological and neural mechanisms underlying the mode of action of other such phyto-extracts with the goal of developing potent but less harmful pest control formulations.
... Momordica charantia, is a tropical to sub-tropical vine in the family Cucurbitaceae. The plant produces triterpenoid compounds in the leaves, fruits and seeds, which have been reported to have antifeedant and oviposition-inhibition properties on arthropods [51][52][53][54] . However, the impact of these biorational insecticides is not well understood on the whitefly and its parasitoid in Bt cotton. ...
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The toxicity of seven biorational insecticides [five insect growth regulators (Buprofezin, Fenoxycarb, Pyriproxyfen, Methoxyfenozide, and Tebufenozide) and two oil-extracts of neem and bitter gourd seeds] against Bemisia tabaci and their selectivity for its parasitoid, Encarsia formosa were evaluated in laboratory and field conditions for 2 years (2018–2019) in Pakistan. Toxicity results demonstrate that Pyriproxyfen, Buprofezin, and Fenoxycarb proved to be effective (80–91% mortality and 66.3–84.2% population-reduction) against B. tabaci followed by Methoxyfenozide, Tebufenozide (50–75% mortality and 47.8–52.4% population-reduction), and then oil-extracts of neem and bitter gourd (25–50% mortality and 36.5–39.8% population-reduction) in the laboratory [72 h post-application exposure interval (PAEI)] and field trails (168 h PAEI), respectively. All tested biorationals, except Methoxyfenozide [(slightly-harmful/Class-II), i.e., causing mortality of parasitoids between a range of 25–50%] and Tebufenozide [(moderately-harmful/Class-III), i.e., causing mortality of parasitoids between the ranges of 51–75%], proved harmless/Class-I biorationals at PAEI of 7-days in the field (parasitism-reduction < 25%) and 3-days in the lab (effect < 30%). In laboratory bioassays, exposure of parasitized-pseudopupae and adult-parasitoids to neem and bitter gourd oils demonstrated that these compounds proved harmless/Class-I biorationals (< 30% mortality). Alternatively, Pyriproxyfen, Buprofezin, Fenoxycarb, Methoxyfenozide, and Tebufenozide were slightly-harmful biorationals (30–79% mortality) against the respective stages of E. formosa. We conclude that most of the tested biorationals proved harmless or slightly harmful to E. formosa, except tebufenozide after PAEI of 7-days (168 h) in the field and, therefore, may be used strategically in Integrated Pest Management (IPM) of B. tabaci.
... Additionally, momordicin I and momordicin II, have exhibited anti-feedant and larvicidal activity in insects (Ling et al., 2008;Li et al., 2015). Moreover, terpenoid compounds including, momordicside -L and momordicoside-K, have demonstrated significant mortality in Armyworm larvae (Wardhani, Abadi, and Himawan, 2015). Furthermore, bioactive compounds of Momordica charantia including, apigenin and tannins have established potential anthelmintic effect (Vedamurthy et al., 2015). ...
Conference Paper
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Mosquito borne diseases contribute to the major disease burden around the world. The use of botanicals as an alternative to synthetic compounds have been explored due to its multifarious advantages. The present study investigated the larvicidal activity of Momordica charantia methanol leaf extracts against the second instar larvae of the laboratory reared, Anopheles tessellatus. Momordica charantia is a medical herb, belonging to the Cucurbitaceae family. The bioassay test was carried out by using the WHO procedure. The mean percentage mortality of Anopheles tessellatus was shown to increase with increasing concentration and increased time of exposure to the Momordica charantia methanol leaf extract. 100% larval mortality was observed at 48 hours following the exposure to plant extract at the highest concentration of 1000 ppm. The study revealed an LC50 value which decreased with time (251 ppm at 24 hours and 126 ppm at 48 hours). Additionally, behavioural changes during the larvicidal assay were observed in response to tactile stimuli. In order to identify histological changes in the cuticle and mid-gut upon exposure to Momordica charantia plant chemicals, the tissue was processed and sections were stained with Hematoxylin and Eosin. The most commonly observed characteristic changes in treated Anopheles tessellatus larvae include damage to cuticle and shrinkage of cells. Therefore, the present investigation revealed that Momordica charantia methanol extract demonstrated effective larvicidal properties against Anopheles tessellatus larvae which can be attributed to the phytochemicals present in the plant. Hence, the formulation of Momordica charantia methanol extracts may potentially be used as an effective and eco-friendly larvicide, which could be an alternative to malaria control.
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The antioxidant and free radical scavenging activities of Bitter Melon Momordica charantia exracts including phenolic compounds, ethanolic and aqueous were studied. Phenolic compounds were extracted, purified and identified by High Performance Liquid Chromatography (HPLC) method. The main phenolic constituents, which were present in the fruit extract of bitter melon, were gallic acid, protocatechuic acid, gentisic acid, catechin, chlorogenic acid and epicatechin. The results clearly indicated that phenolic compounds have an effective antioxidant activity by using Ferric Thiocyanate (FTC) method. Phenolic compounds caused 91.25% lipid peroxidation inhibition of linoleic acid emulsion. This activity was greater than ethanolic extract 82.5%, α-tocopherol 70% and aqueous extract 49.58%. Also the phenolic compounds revealed obvious activity for HR 2 ROR 2 Rscavenging 68.8% in comparison with α-tocopherol 45.3%, ethanolic extract 52.6% and aqueous extract 36.2%. These results confirmed the important role of phenolic compounds as antioxidants and the most antioxidant activity of bitter melon fruits belong to these compounds. ‫ﺍﻟﻤﺴﺘﺨﻠﺺ‬ ‫ﺍﻟﻜﻴﺮﻳﻼ‬ ‫ﺛﻤﺎﺭ‬ ‫ﻟﻤﺴﺘﺨﻠﺼﺎﺕ‬ ‫ﺍﻟﺤﺮﺓ‬ ‫ﻟﻠﺠﺬﻭﺭ‬ ‫ﺍﻟﻤﺰﻳﻠﺔ‬ ‫ﻭﺍﻟﻔﻌﺎﻟﻴﺔ‬ ‫ﻟﻼﻛﺴﺪﺓ‬ ‫ﺍﻟﻤﻀﺎﺩﺓ‬ ‫ﺍﻟﻔﻌﺎﻟﻴﺔ‬ ‫ﺩﺭﺳﺖ‬ Momordica charantia ‫ﺍﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﺮﻛﺒﺎﺕ‬) ، (‫ﻭﺍﻟﻤﺎﺋﻲ‬ ‫ﺍﻻﻳﺜﺎﻧﻮﻟﻲ‬ ، ‫ﻭﺷﺨﺼﺖ‬ ‫ﺍﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﺮﻛﺒﺎﺕ‬ ‫ﻭﻧﻘﻴﺖ‬ ‫ﺃﺳﺘﺨﻠﺼﺖ‬ ‫ﻛﻤﺎ‬ ‫ﻣﺴﺘﺨﻠﺺ‬ ‫ﻓﻲ‬ ‫ﺍﻻﺳﺎﺳﻴﺔ‬ ‫ﺍﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﻜﻮﻧﺎﺕ‬ ‫ﺷﺨﺼﺖ‬. ‫ﺍﻷﺩﺍء‬ ‫ﻋﺎﻟﻴﺔ‬ ‫ﺍﻟﺴﺎﺋﻠﺔ‬ ‫ﺍﻟﻜﺮﻭﻣﺎﺗﻮﻛﺮﺍﻓﻴﺎ‬ ‫ﻁﺮﻳﻘﺔ‬ ‫ﺑﺎﺳﺘﺨﺪﺍﻡ‬ ‫ﺍﻟﻜﺎﻟﻚ‬ ‫ﺣﺎﻣﺾ‬ ‫ﻣﺜﻞ‬ ‫ﺍﻟﻜﻴﺮﻳﻼ‬ ‫ﺛﻤﺎﺭ‬ ، ‫ﺍﻟﺒﺮﻭﺗﻮﻛﺎﺗﺠﻮﻳﻚ‬ ‫ﺣﺎﻣﺾ‬ ، ‫ﺍﻟﺠﻨﺘﺴﻚ‬ ‫ﺣﺎﻣﺾ‬ ، ‫ﺍﻟﻜﺎﺗﺠﻴﻦ‬ ، ‫ﺣﺎﻣﺾ‬ ‫ﻭﺍﻻﻳﺒﻴﻜﺎﺗﺠﻴﻦ‬ ‫ﺍﻟﻜﻠﻮﺭﻭﺟﻨﻚ‬ ‫ﺃﺷﺎ‬. ‫ﺍﻟﻜﻴﺮﻳﻼ‬ ‫ﺛﻤﺎﺭ‬ ‫ﻣﺴﺘﺨﻠﺺ‬ ‫ﻓﻲ‬ ‫ﺍﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﺮﻛﺒﺎﺕ‬ ‫ﺍﻥ‬ ‫ﺍﻟﻰ‬ ‫ﻭﺍﺿﺢ‬ ‫ﺑﺸﻜﻞ‬ ‫ﺍﻟﻨﺘﺎﺋﺞ‬ ‫ﺭﺕ‬ ‫ﺍﻟﺤﺪﻳﺪﻳﻚ‬ ‫ﺛﺎﻳﻮﺳﻴﺎﻧﺎﺕ‬ ‫ﻁﺮﻳﻘﺔ‬ ‫ﺑﺄﺳﺘﺨﺪﺍﻡ‬ ‫ﻟﻸﻛﺴﺪﺓ‬ ‫ﻣﻀﺎﺩﺓ‬ ‫ﺷﺪﻳﺪﺓ‬ ‫ﻓﻌﺎﻟﻴﺔ‬ ‫ﺃﻣﺘﻠﻜﺖ‬ ‫ﺍﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﺮﻛﺒﺎﺕ‬ ‫ﺛﺒﻄﺖ‬. ‫ﺑﻨﺴﺒﺔ‬ ‫ﺍﻟﻠﻴﻨﻮﻟﻴﻚ‬ ‫ﺣﺎﻣﺾ‬ ‫ﻟﻤﺴﺘﺤﻠﺐ‬ ‫ﺍﻟﺪﻫﻮﻥ‬ ‫ﺃﻛﺴﺪﺓ‬ ‫ﺍﻟﻤﺴﺘﺨﻠﺼﺔ‬ 91.25 ‫ﺍﻷﻳﺜﺎﻧﻮﻟﻲ‬ ‫ﺍﻟﻤﺴﺘﺨﻠﺺ‬ ‫ﻣﻦ‬ ‫ﺃﻋﻠﻰ‬ ‫ﻭﻫﻮ‬ % 82.5 ‫ﻭﺍﻷﻟﻔﺎﺗﻮﻛﻮﻓﻴﺮﻭﻝ‬ % 70 ‫ﺍﻟﻤﺎﺋﻲ‬ ‫ﻭﺍﻟﻤﺴﺘﺨﻠﺺ‬ % 49.58 % ، ‫ﻓﻌﺎﻟﻴﺔ‬ ‫ﺍﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﺮﻛﺒﺎﺕ‬ ‫ﺃﻅﻬﺮﺕ‬ ‫ﻛﺬﻟﻚ‬ ‫ﻭﺑﻨﺴﺒﺔ‬ ‫ﺍﻟﻬﻴﺪﺭﻭﺟﻴﻦ‬ ‫ﺑﻴﺮﻭﻛﺴﻴﺪ‬ ‫ﻷﺯﺍﻟﺔ‬ ‫ﻭﺍﺿﺤﺔ‬ 68 ‫ﺑﺎﻷﻟﻔﺎﺗﻮﻛﻮﻓﻴﺮﻭﻝ‬ ‫ﻣﻘﺎﺭﻧﺔ‬ % 45.3 ‫ﻭﺍﻟﻤﺴﺘﺨﻠﺺ‬ % ‫ﺍﻷﻳﺜﺎﻧﻮﻟﻲ‬ 52.6 ‫ﺍﻟﻤﺎﺋﻲ‬ ‫ﻭﺍﻟﻤﺴﺘﺨﻠﺺ‬ % 36.2 ‫ﺍ‬ ‫ﻟﻠﻤﺮﻛﺒﺎﺕ‬ ‫ﺍﻟﻤﻬﻢ‬ ‫ﺍﻟﺪﻭﺭ‬ ‫ﺍﻛﺪﺕ‬ ‫ﺍﻟﻨﺘﺎﺋﺞ‬ ‫ﻫﺬﻩ‬. % ‫ﻛﻤﻮﺍﺩ‬ ‫ﻟﻔﻴﻨﻮﻟﻴﺔ‬ ‫ﺍﻟﻤﺮﻛﺒﺎﺕ‬ ‫ﻟﻬﺬﻩ‬ ‫ﺗﻌﻮﺩ‬ ‫ﺍﻟﻜﻴﺮﻳﻼ‬ ‫ﻧﺒﺎﺕ‬ ‫ﺛﻤﺎﺭ‬ ‫ﻓﻲ‬ ‫ﻟﻼﻛﺴﺪﺓ‬ ‫ﺍﻟﻤﻀﺎﺩﺓ‬ ‫ﺍﻟﻔﻌﺎﻟﻴﺔ‬ ‫ﻣﻌﻈﻢ‬ ‫ﻭﺍﻥ‬ ‫ﻟﻼﻛﺴﺪﺓ‬ ‫ﻣﻀﺎﺩﺓ‬ .
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The antioxidant and free radical scavenging activities of Bitter Melon Momordica charantia exracts including phenolic compounds, ethanolic and aqueous were studied. Phenolic compounds were extracted, purified and identified by High Performance Liquid Chromatography (HPLC) method. The main phenolic constituents, which were present in the fruit extract of bitter melon, were gallic acid, protocatechuic acid, gentisic acid, catechin, chlorogenic acid and epicatechin. The results clearly indicated that phenolic compounds have an effective antioxidant activity by using Ferric Thiocyanate (FTC) method. Phenolic compounds caused 91.25% lipid peroxidation inhibition of linoleic acid emulsion. This activity was greater than ethanolic extract 82.5%, - tocopherol 70% and aqueous extract 49.58%. Also the phenolic compounds revealed obvious activity for H2O2 scavenging 68.8% in comparison with - tocopherol 45.3%, ethanolic extract 52.6% and aqueous extract 36.2%. These results confirmed the important role of phenolic compounds as antioxidants and the most antioxidant activity of bitter melon fruits belong to these compounds.
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Bitter melon, Momordica charantia L, is a popular traditional medicinal fruit in tropical and subtropical countries. It has been linked with therapeutic effects, some of which are likely due to its flavonoids. To determine its total flavonoid content (TFC) and to prepare extracts for use as nutritional supplements or ingredients for nutraceutical functional foods, various solvents have been used, including water, which is the preferred solvent because it is inexpensive, safe and environmentally friendly. The study aimed to extract bitter melon, using five solvents (ethanol, methanol, n-butanol, acetone and water) before and after the optimal conditions for water were determined in terms of extraction temperature, time, ratio of water to bitter melon (mL/g) and number of times the same material was extracted. The TFC of six varieties of bitter melon was also determined. Acetone was the best of the five solvents for extracting flavonoids from the Moonlight variety (23.2 mg Rutin Equivalents (RE)/g). Even after increasing the extraction by 88% (1.24 vs 0.66 mg RE/g) using optimised conditions for the aqueous extraction (two extractions at 40˚C for 15 min at a ratio of 100:1 mL/g of bitter melon powder), the fla-vonoids extracted from the Moonlight variety using water was very little (5.4%) compared to acetone. Furthermore , using acetone, it was shown that the Moonlight variety (23.2 mg RE/g) bought at a local market had higher levels of flavonoids than the greenhouse-grown Jade (15.3 mg RE/g), Niddhi (16.9 mg RE/g), Indra (15.0 mg RE/ g), Hanuman (3.9 mg RE/g) and White (6.9 mg RE/g) varieties. Therefore, acetone was the best solvent for extracting flavonoids from bitter melon and the aqueous extraction could only be improved to extract 5.4% of the flavonoids extracted with acetone from the Moonlight variety, which had the highest TFC of the six varieties of bitter melon.
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Plants with potential therapeutic values have been used from time immemorial to cure various ailments and infectious diseases. Of late, scientific evidences have been provided on the potential therapeutic agent exhibited by certain traditionally used vegetable extracts. The importance of wild edible plants may be traced to antiquity but systemic studies are recent. All the Momordica species have been consumed as vegetable and traditionally used for various disorders. The whole plant parts are ascribed to possess the anti-diabetic effect in traditional medicinal system. The active constituents of Momordica plant parts were cucurbitane type triterpenoids, phenolics, glycosides, and several kinds of peptides including Momordica anti-HIV protein (MAP 30). Recent reports revealed the presence of several kinds of cucurbitane type triterpenoids in leaf, stem and fruits of Momordica species having several pharmacological activities. There is lack of scientific information available on the wild species which also having several bioactive components with potential activities. So the present review compares and highlights the current knowledge of the nutritional value, phytochemistry and physiological effects of wild species with known variety.
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There are many methods to manage plant pests and diseases. One method that is always used by farmers is chemical control using synthetic pesticides. Utilization of synthetic pesticides inappropriately in terms of kind, target, dose/concentration, technique and time can be unsafe to the environment as well as the consumers. The objective of this research was to determine pesticide residues on fresh vegetables in the central market of Ambon city. The vegetables being sampled were spinach (Amarantus indica), water cresant (Ipomoea aquatica), green mustard (Brassica juncea) and long bean (Vigna sinensis) collected from Mardika and Passo local markets, Ambon City. Residue analysis was conducted at the Testing Laboratory of the Agricultural Post Harvest Research and Development Board of the Ministry of Agriculture, Bogor (Laboratorium Pengujian, Balai Besar Penelitian dan Pengembangan Pasca Panen Pertanian, Kementrian Pertanian, Bogor). The results showed that the residues of Organochlorine (Chlorinated Hydrocarbons), Organophosphate, Carbamate and Pyrethroid were detected on sampled vegetables from central marketing at Ambon City. The residues of these classes of synthetic pesticides were detected in the form of Heptachlor, Aldrin, Endosulfan, Lindane, Chlorpyrifos, Profenofos, Diazinon, Monocrotophos, Parathion, Carbofuran, and Cypermethrin. The residues detected were below Maximum Residue Limit (MRL) based on SKB of the Minister of Health and the Minister of Agriculture (Menteri Kesehatan dan Menteri Pertanian) No. 881/MENKES/SKB/VIII/1996 and No. 711/Kpts/TP270/8/96, and The Regulation of the Minister of Agriculture (Peraturan Menteri Pertanian) No.27/Permentan/ PP.340/5/2009
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Bitter gourd, Momordica charantia, was less palatable to 2 species of armyworms, Spodoptera litura and Pseudaletia separata, than 2 other cucurbitaceous plants. A methanol extract of M. charantia leaves inhibited feeding of the armyworm larvae. The 2 most active fractions obtained by silicagel chromatography were purified by HPLC. Momordicine II, triterpene mono-glucoside, was identified as an antifeedant compound from the most active of these fractions. The second active fraction led to the isolation of a new triterpene di-glucoside. Fresh leaves of M. charantia contained ca. 0.3% of momordicine II. Momordicine II showed a significant antifeedant effect on P. separata at the concentrations of 0.02, 0.1 and 0.5% in artificial diets. Momordicine II caused a significant feeding reduction in S. litura only at the highest concentration (0.5%) tested. The difference in the feeding response of the 2 armyworms to momordicine II may be related to the diversity in their host range. The author also examined whether stress applied to plant exerted an effect on insect's feeding preference. M. charantia is the host plant for the larvae of the pyralid moth, Diaphania indica but not for those of P. separata. Feeding response to UV-irradiated M. charantia leaves was compared between these 2 insects. D. indica preferred intact leaves, while P. separata preferred UV-irradiated leaves. These differences might be caused by the difference in the contents of antifeedants and feeding stimulants in the intact and UV-irradiated leaves.
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Background & objectives: Development of insect resistance to synthetic pesticides, high operational cost and environmental pollution have created the need for developing alternative approaches to control vector-borne diseases. In the present study, larvicidal activity of flavonoid extracts of different parts of Vitex negundo (Linnaeus) and Andrographis paniculata (Nees) have been studied against the late III or early IV instar larvae of Aedes aegypti and Anopheles stephensi (Liston). Methods: Flavonoids were extracted from different parts of the selected plants using standard method. Bioassay test was carried out by WHO method for determination of larvicidal activity against mosquitoes. Different compounds of the most active extract were identified by the gas chromatography-mass spectrometry (GC-MS) analysis. Results: Flavonoid extract of whole aerial part of A. paniculata was found to be inactive against the selected larvae of Ae. aegypti even at the concentration of 600 ppm, whereas it caused 70% mortality in An. stephensi at the concentration of 200 ppm. Flavonoid extract of flower-buds produced highest mortality (100%) at the concentration of 600 ppm for the late III or early IV instar larvae of Ae. aegypti and at the concentration of 200 ppm for the larvae of An. stephensi. GC-MS analysis of the most active flavonoid extract from flower-buds of Vitex showed 81 peaks. Phenol (26.83% area), naphthalene (4.95% area), 2,3-dihydrobenzofuran (6.79% area), Phenol-2,4-Bis (1,1-dimethyl) (4.49% area), flavones 4'-OH,5-OH,7-di-O-glucoside (0.25% area) and 5-hydroxy- 3,6,7,3',4'-pentamethoxy flavones (0.80% area) were present in major amount. Conclusion: Flavonoid extracts from different parts of two selected plants possess larvicidal activity against two selected mosquito species, hence, could be utilized for developing flavonoid-based, eco-friendly insecticide as an alternative to synthetic insecticides.