4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

BIOACTIVITIES OF JATROPHA CURCAS LINN LATEX

Mohamad Syakir M.S.1, Ismail J.1, Zaini A.2, Nur Diyana I.1
1
Department of Plant Science and Environmental Ecology
2
Department of Chemistry
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak (MALAYSIA)
e-mails: syakir.sarib@gmail.com

Abstract
Jatropha curcas L. is a perennial tree under Euphorbiaceae family and well known for alternative
source of biodiesel obtained from its seed. The study examined the latex of J. curcas crude that might
be toxic to human and plant pathogen fungi namely Fusarium oxyporum, F. solani and Aspergillus
niger; wood decay fungi which were Trametes versicolor and Gleophylum trabeum. The specific
objectives of this study were firstly to determine the toxicity of crude powdered J.curcas latex,
secondly to determine the chemical compositions of crude powdered J. curcas latex and thirdly to
determine the chemical constituents of crude powdered J.curcas latex. Three different concentrations
of J. curcas latex (100 mg/ml, 50mg/ml and 25 mg/ml) were prepared for in vitro bioassay test using
agar well diffusion method. The wells were filled with concentration prepared and the growths of
microorganisms were observed after 72 hours incubation period. The crude latex of J.curcas was
subjected to composition and constituent analyses using Gas Chromatography – Mass Spectroscopy
(GC/MS). In toxicity test, all tested fungi showed significant inhibition to all fungus tested except A.
niger. The main compounds detected were dotriacontane (24%), pentatriacontane (20%),
hexatriacontane (20%), 1,2-benzenedicarboxylic acid (41%) and -sitosterol (35%). Further study
should be carried out to tap the potential of J. curcas latex as active ingredient in the pharmaceutical
and pesticide production.
Keywords: Jatropha curcas; pure powdered latex; toxicity; chemical composition; chemical constituent.

1 INTORDUCTION
Jatropha curcas Linn. belongs to the family Euphorbiaceae, known as ‘Jarak Pagar’ in Malay and
widely known as physic nut. It is a drought resistant and perennial tropical plant that can be grown in
low to high rainfall areas either in the farms as a commercial crop or on the boundaries as a hedge to
protect fields from grazing animals and to prevent erosion (Irvine, 1961). It grows under a wide range
of rainfall regimes from 250 to over 1200 mm per annum (Katwal & Soni, 2003). The extract of its
leaves has antifungal properties (Garcia & Lawas, 1990). Fruit of J. curcas is highly toxic and may
lead to death if consumed. According to Goonasekera et al. (1995), the fruit may cause pregnancy-
terminating in rat which benefit the pest management industry. However, the toxicity can be removed
and utilized to many other purposes. The water extract of J. curcas branches showed inhibition on the
HIV induced cytopathic effect with low cytotoxicity (Matsuse et al., 1999).

1.1 Jatropha curcas Latex

The latex of J. curcas obtained from its branches reported to contains an alkaloid known as jatrophine,
which is believed to be having anti-cancerous properties (Henning, 2003). The healing effect of
curcain a proteolytic enzyme from the latex on wound has been demonstrated (Nath & Dutta, 1991).
The latex combined with the powdered leaves is applied to sluggish wounds while when formulated as
enema it is used for the treatment of gonorrhoea (Irvine, 1961). The latex also used in healing of
wounds, refractory ulcers, and septic gums and as a styptic in cuts and bruises. A proteolytic enzyme
(curcain) has been reported to have wound healing activity in mice (Nath & Dutta, 1997). The latex is
used to treat fungal infections in the mouth, bee and wasp stings and digestive problems of children in
Mexico (Schmook & Serralta-Peraza, 1997). The chemical composition of J. curcas latex was reported
to contain curcacycline A that has anti tumour properties (Van den Berg et al., 1995) and curcain, a
protease (Nath & Dutta, 1991). Previous study on the latex using IR spectrum of ethyl acetate extract
reported the presence of aromatic phenolic compounds and phenolic compounds which generally
behave as acids that have high antimicrobial activity (Gisvold, 1977). In phytochemical screening
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

study done by Suhaili et al. (2011) on the crude latex extract revealed the presence of saponins and
tannins as had previously been reported on other parts of the plant. Saponins and tannins, in
particular, have been reported to possess antimicrobial activity (Zakaria et al., 2010)

2 METHODOLOGY

2.1 Latex Collection

The fresh latex of J. curcas was collected from Jatropha orchard owned by Carbon Capital Coperation
in Sadong Jaya, Kota Samarahan. Latex were collected and stored into clean sterilized bottle warped
with aluminum foil to prevent phytochemical reaction. Sample collected were then directly brought into
the laboratory and stored in 4°C.

2.2 Drying and Powdering of Latex

Latex was spread on sterilized glass Petri dish and kept in fume cupboard in dark condition for 24
hours. Dried latex were subsequently scrapped off carefully using sterilized glass slide and mashed to
obtain finer powder. The powder was then sterilized using UV light inside laminar flow for 10 minutes.

2.3 Fungi Preparation

Trametes versicolor, Aspergillus niger, Fusarium oxysporum, F. solani, and Gloeophylum trabeum
cultured on Malt Extract Agar (MEA) was used for culturing all the fungi. 15ml of MEA solution was
poured into sterile disposable Petri dish and left cools until it solidified. Fungi were then inoculated
from stock culture onto agar plate and were re-inoculated after one week to obtain pure culture of
fungus.

2.4 Toxicity test

The agar well diffusion method adapted from Suhaili et al. (2011) and Oyi et al. (2007) was employed
in this study with several modifications. Three well were made using cork borer (10 mm diameter) in
25 ml agar. Three different concentrations of J.curcas latex diluted using metanol (25 mg/ml, 50
mg/ml, 25 mg/ml) filled into well, each 30µl and left for 20 minutes. Plug of test fungi (± 5mm X 5mm)
from cultured stock were next placed on the centre of solidified agar. Then it was incubated at 30 ºC
and the data were recorded on 5th day after incubation. A plate with single well was prepared and filled
with 30µl methanol for each fungi tested as a control. Antifungal activities were calculated by
measuring the zone of inhibition or clear zone. Inhibition zones above 20 mm were classified as
“strong”, 19 mm to 15 mm as “moderate” and below 14mm as “weak” activities.

2.5 Fractionation and Chemical Analysis

Column chromatography was employed to fractionate the pure powdered Jatropha curcas latex using
19 solvent systems with different ratio of n-hexane, dichloromethane, ethyl acetate and metanol. The
fractions were then subjected to Gas Chromatography equiped with Mass Spectrometer. Identification
of chemical components was made based on comparison with GC retention times of standard
references materials. It was also assisted by comparing the mass spectra obtained during analysis
with those mass spectra stored in the National Institute of Standard and Technology (NIST) standard
library incorporated in the GC/MS data system.

3 RESULTS AND DISCUSSION

3.1 Toxicity Test

The toxicity tests of crude powdered latex of J. curcas were carried out by using 100mg/ml, 50 mg/ml
and 25 mg/ml concentrations diluted in methanol. From the test, it was observed that J.curcas latex
has toxicity towards T. versicolor, G. trabeum, F. oxysporum, F. solani in all concentrations of crude
powdered J.curcas latex applied. Fusarium solani was recorded to have the least inhibition for all
concentrations of powdered J.curcas latex. Fig. 1 shows the example for the formation of inhibition
zone against F. solani with different concentrations of crude J. curcas latex observed from toxicity test.
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

However, A. niger showed no significant effect of J.curcas latex on the growth of this fungus in any
concentrations of J. curcas latex (Fig. 2).

100 mg/ml concentration showed strong inhibition with inhibition diameter more than 20 mm.
Gleophylum trabeum was observed to strongly inhibited with 27 mm inhibition zone. F. Oxysporum
and T. versicolor recorded 25.7 mm zone of inhibition. F. solani formed inhibition zone of 20 mm. For
50 mg/ml concentration, inhibition zone of T. versicolor was 22 mm, indicated strong inhibition while
the lowest inhibition diameter was F. solani with 16 mm indicated moderate activity. Both F.
oxysporum and G. trabeum showed inhibition zone of 21 mm. For 25 mg/ml concentration, inhibition
zone was strong on both F. oxysporum and T. versicolor with 20 mm while less inhibition was
observed on F. solani with 14 mm which indicated moderate activity. G. trabeum was observed to
inhibited by 19 mm zone of inhibition which indicated moderate inhibition.

Figure 1: Formation of inhibition zone against Figure 2: No inhibition zone showed against
Fusarium solani. Aspergillus niger.

Table 1 summarize zone of inhibitions of J.curcas latex and the readings were taken as mean of three
replicates. Fig. 3 visualized the inhibition classification of different types of fungi and its concentration
and shows different fungi has different inhibition strength for each concentration. Pure latex of
J.curcas has toxicity affect on several human and plant pathogen fungi. Study done by Schmook and
Serralta-Peraza (1997) proved that J. curcas latex can be used to treat fungal infection Henning
(2003) analyzed the presence of alkaloid in J. curcas called jatrophine and it’s believed to have anti
dermatomucosal disease. Ambuse & Bhale (2012) reported J. curcas latex showed toxicity toward the
growth of Fusarium proliferatum and F. pytium. It also been reported that the crude extracts of ethyl
acetate and methanol from stem bark has no significant affect towards A. niger (Gupta et al., 2010).

Table 1: Inhibition zones of fungal species in agar well diffusion plate assay of powdered J. curcas
latex after 5 days incubation
Conc. of latex (mg/ml) Fungi / Zone of inhibition (mm)
FS FO TV GT AN
100 20 ± 2.3 25.7 ± 0.3 25.7 ± 0.8 27 ± 1.5 -
50 16.3 ± 1.2 21.7 ± 0.3 22 ± 1.2 21.7 ± 0.9 -
25 14 ± 1.2 20 ± 0.6 20 ± 2.3 19 ± 1.7 -
Methanol - - - - -

FS = Fusarium solani; FO = Fusarium oxysporum; TV = Trametes versicolor; GT = Gleophylum trabeum; AN =

Aspergillus niger; - = No inhibition
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

Fig3: Inhibition classification of three concentrations of J. curcas latex against tested fungi

3.2 Chemical Constituents and Compound Analysis

The GC/MS analyses revealed that the most abundantce compounds in hexane and methanol
fractions were dotriacontane and 1,2-benzenedicarboxylic acid.
3.2.1 n-hexane Fraction
Major chemical constituents of hexane fraction of J.curcas latex was dotriacontane with percentage of
composition 24% (Fig 4). Other significant compounds were pentatriacontane (20%), hexatriacontane
(19%), tetracosane (19%), tetratricontane (10%) and nonacosane (5%) (Table 2).
The chemical constituents in hexane fraction contain nonacosane was reported to responsible in the
inhibition of several types of bacterial formation (Yayli et al, 2006). Nonacosane is known as a plant
compound that responsible in formation many of waxy layer of long chain paraffin (Hankin &
Kolattukudy, 1968).

Fig 4: Gas chromatogram traced by GC/MS for n-hexane fraction of powdered J.curcas latex

Tetratriacontane was also reported in the extract of Dictyopteris membracea and showed significants
for antimicrobial activities (Ozdemir et al, 2006). Dotriacontane was reported to show toxicity affect on
the microbe tested. (Amin & Sleem, 2007). This compound was also found in extract of Sideritis
scardica and it showed antimicrobial properties (Vanja et al., 2012). Hexatriacontane is classified as
flavanoid and has very high medicinal value (Vaishali et al., 2009). Yassa et al. (2009) reported
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

Hexatriacontane can be used as active preventing agent for many diseases and also has antioxidant
affects. Tetracosane was also reported in flower extract of Allium atroviolaceum and showed positive
antibacterial activity (Dehpour et al., 2011). It also found in the essential oil of Geranium columbinum
that has toxicity affects (Radulovic et al., 2011)

Table 2: Major chemical constituents identified in n- hexane fraction of pure latex of J.curcas

3.2.2 Methanol Fraction

Methanol fraction is assumed to consist of intermediate polar compounds such as fatty acids and most
abundance chemical identified was mainly classified in alcohol group. Figure 5 shows presence of 1,2-
benzenedicarboxylic acid (40.96%), ß-Sitosterol (35.06%), 2-Hexyl-1-decanol (9.30%) and 4,6-
Cholestadien-3.beta.-ol (4.72%) (Table 3).

2-hexyl-decanol was reported to has antibacterial activities (Najiah et al,. 2008) and also found in Thai
honey bee which has antibiotic affect for many human diseases (Suwannapong et al., 2011). 1,2-
benzenedicarboxylic acid is a plasticizer compound reported to have high toxicity and antimicrobial
effect against Listeria (Hsouna, 2011) and reported as the active phytochemical compound (Chen et
al., 2011). Compound 4,6-Cholestadien-3.beta.-ol was found in extract of Azadiracta indica which had
showed antibacterial activity (Moorthy & Boominathan, 2011).

Fig 5: Gas chromatogram traced by GC/MS for methanol fraction of powdered J.curcas latex
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

Compound ß-Sitosterol was also been found in the root and kernel meal of J.curcas in phenolics and
flavonoids analyses by HPLC analysis (Oskoueian et al., 2011). The compound was also found in the
leaves of J. curcas. ß-Sitosterol has no antifungal effect (Hegazy et al, 2010), but it had significant
effect on bacteria (Gohari et al., 2009)

Table 3: Major chemical constituents identified in methanol fraction of pure latex of J.curcas

4 CONCLUSION
The toxicity test showed J. curcas latex inhibited Trametes versicolor, Gleophyllum trabeum, Fusarium
oxysporum, Fusarium solani, but negative to Aspergillus niger indicate the powdered latex has limited
inhibition affect different type of organisms.The main compounds detected include dotriacontane,
pentatriacontane, hexatriacontane, 1,2-benzenedicarboxylic acid and -sitosterol. Extensive research
need to be done with various concentrations of Jatropha curcas latex and organisms. Extraction of
pure compounds to obtain significant active ingredient and finding which inert ingredients have the
potential to be amplified for chemical activation, will lead to the diversification of J. curcas utilization,
and hence benefit industries related to Jatropha.

REFERENCES
Ambuse, M. G., & Bhale, U. N. (2012). Evaluation of antifungal activity of plant latex extracts against
resistant isolates of pathogens associated on Rumex Acetosa L. International Journal Of
Ayurvedic And Herbal Medicine, (2) 2:389-393
Amin, W. M. A., & Sleem, A. A. (2007). Chemical and biological study of aerial parts of Anethum
graveolens L. Egypt Journal of Biomedical Science, (23) 13: 188-190
Bruce, A. & Highley, T. L. (1991). Control of growth of wood decay Basidiomycetes
by Trichoderma spp. and other potentially antagonistic fungi. Forest Product Journal, 41: 63-
67.
Chen, H., Yang, Y., Xue, J., Wei, J., Zhang, Z., & Chen H. (2011). Comparison of compositions and
antimicrobial activities of essential oils from chemically stimulated agarwood, wild agarwood
and healthy Aquilaria sinensis (Lour.) Gilg trees. Molecules Journal. (16): 4884-4896
Dehpour, A. A., Babakhani, B., Khazaei, S., & Asadi, M. (2011). Chemical composition of essential oil
and antibacterial activity of extracts from flower of Allium atroviolaceum. Journal of Medicinal
Plants Research, (5) 16: 3667-3672
Garcia, R.P., & Lawas, P. (1990). Note: Potential plant extracts for the control of Azolla fungal
pathogens. Philippine Agriculture, (73) 4: 343-348
Gisvold, O. (1977). Phenols and their derivatives. In: Textbook of organic Medicinal and
Pharmaceutical Chemistry 7th Edition by Wilson, C. O., Gisvold, O & George, R. F. (1977).
Philadephia.: J.B Lippincott company, pp. 182-184.
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

Gohari, A. R., Saeidnia, S., Shaverdia, A. R., Yassa, N., Malmir, M., Mollazade, K., & Naghinejad, A.
R. (2009). Phytochemistry and antimicrobial compounds of Hymenocrater calycinus. EurAsia
Journal of BioScience, (3): 64-68
Goonasekera, M. M., Gunawardana, V. K., Jayasena, K., Mohammed, S. G., & Balsubramaniam, S.
(1995). Pregnancy terminating effect of Jatropha curcas in rats. Journal of
Ethnopharmacology, (47) 3:117–123
Gupta, D. D., Haque, M. E., Islam, M. N., Mondal, M. S. I., & Shibib, B. A. (2010) Antimicrobial and
cytotoxic activities of Jatropha curcas (Euphorbiacea). Dhaka Universities Journal of
Pharmaceutical Sciences, (9) 2: 139-142
Hankin, L., & Kolattukudy, P. E. (1968). Metabolism of a Plant Wax Paraffin (n-Nonacosane) by a Soil
Bacterium (Micrococcus cerificans). Microbiology, 51: 457
Hegazy, A. K., Sayed, A. M., & Kabiel, H. F. (2010). Study on combined antimicrobial activity of some
biologically active constituent from wild Moringa peregrine Forssk. Journal of yeast and
fungal research, (1) 1:15-24
Henning, R.K. (2003). A Guide to Jatropha Promotion in Africa. Weissensberg, Germany: bagani GbR.
Hsouna, A. B., Trigui, M., Mansour, R. B., Jarraya, R. M., Damak, M., & Jaoua, S. (2011). Chemical
composition, cytotoxicity effect and antimicrobial activity of Ceratonia siliqua essential oil with
preservative effects against Listeria inoculated in minced beef meat. International Journal of
Food Microbiology, (148) 1: 66-72
Irvine, F. R. (1961). Woody Plants of Ghana 1st Edition. London: Oxford University Press.
Katwal, R. P. S., Soni, P. L. (2003). Biofuels: an opportunity for socioeconomic development and
cleaner environment. Indian Forester, (129): 939–949.
Matsuse, I. T., Lim, Y. A., Hattori, M., Correa, M., & Gupta, M.P. (1999). Search of anti-viral properties
in Panamanian medicinal plants, the effects on HIV and its essential enzymes. Journal of
Ethnopharmacology, (64): 15-22.
Moorthy, V., & Boominathan, M. (2011). The antimicrobial activities of crude extracts and fraction of
Psidium guajava and Azadirachta indica against Staphylococcus aureus in chronic disease
affected patients. International Journal of Universal Pharmacy and Life Sciences, (1) 2: 160-
173
Najiah, M., Wei, L. S., Seng, C. T., Wee, W., & Leong L. K. (2008). Potential of edible plants as
remedies of systemic bacterial disease infection. Cultured Fish Global Journal of
Pharmacology 2, (2): 31-36
Nath, L. K., & Dutta, S. K., (1992). Extraction and purification of curcain, a protease from the latex of
Jatropha curcas L. J. Pharm. Pharmacology, (43) 111–114.
Oskoueian, E., Norhani Abdullah, Syahida Ahmad, Wan Zuhainis Saad, Abdul Rahman Omar, & Ho,
Y. W. (2011). Bioactive compounds and biological activities of Jatropha curcas L. kernel meal
extract. International Journal of Molecular Science, (12): 5955-5970
Oyi, A. R. Onaolapo, J.A & Adigun, J. O. (2002). Phytochemical and antimicrobial Screening of the
latex of Jatropha curcas Linn (Euphorbiaceae). Journal of phytomedicine and Therapeutical
(1&2), 63-74.
Ozdemir, G., Horzum, Z., Atakan, S., & Ulku, K. Y. (2006) Antimicrobial activities of volatile and
various component and extracts of Dictyopteris membranacea and Cystoseira barbata from
the coast of Izmir, Turkey. Turkish Journal of Chemistry, (3) 44: 183-188
Radulovic, N., Deki, M., Stojanovi, Z. R., & Pali’c, R. (2011). Chemical composition and antimicrobial
activity of the essential oils of Geranium columbinum L. and G. lucidum L. (Geraniaceae).
Turkish Journal of Chemistry, (35): 499-512
Schmook, B., Serralta, P. L. (1997) Jatropha curcas: distribution and uses in the Yucantan Penisula.
Proceeding of First International Symposium on Biofuel and Industrial Product from Jatropha
curcas and other Tropical Oil Seed Plants, Managua, Nicaragua.
4th Regional Conference on Natural Resources in the Tropics (NTrop4), 2012

Suhaili, Z., Yeo, C. C., Yasin, H. N., Badaludin, N. A., & Zakaria, Z. A. (2011). Antibacterial profile of
Jatropha curcas latex extracts against selected human pathogenic bacteria. African Journal of
Microbiology Research, (5) 29:5147-5154
Suwannapong, G., Benbow, M. E., & Nieh, J. C. (2011). Biology of Thai honeybees: natural history
and threats. Thailand: Nova Science. pp. 1-98.
Vaishali, A., Khatiwora, E., Manik, K., Tambe, A., Pawar, P. & Deshpande, N. (2009). GC-MS study of
fatty acid, ester and alcohol from leaves of Ipomea carnea. International Journal of
Pharmaceutical Technology Reseach, (1) 4: 1224-1226
Van den Berg, A. J., Horsten, S. F., Kettenes van den Bosch, J. J., Kroes, B. H., Beukelman, C. J.,
Loeflang, B. R., & Labadie, R. P., (1995). Curcacycline A: a novel cyclic octapeptide isolated
from the Jatropha curcas. FEBS Letters, (358): 215–218.
Vanja, T., Dragica, B., Ivana, A., Sofija, D., & Ksenija, A. (2012). Chemical and antimicrobial
evaluation of supercritical and conventional Sideritis scardica L. extract. Molecules Journal,
17: 2683-2703
Yassa, N., Masoomi, F., Rohani, R. S. E., & Hadjiakhoondi, A. (2009). Chemical composition and
antioxidant activity of the extract and essential oil of Rosa damascena from Iran, population of
Guilan. DARU Journal of Pharmaceutical Sciences, (17) 3: 175-179
Yayli, N., Conan, G. Osman, U. C., Ahmet, Y., Ulker, S., Kamil, C., & Salih, T. (2006). Composition
and Antimicrobial Activities of Volatile Components of Minuartia meyeri. Turkish Journal of
Chemistry, 3: 71-76
Zakaria, Z. A., Sufian, A. S., Ramasamy, K., Ahmat, N., Sulaiman, M. R., Arifah, A. K., Zuraini, A.,
Somchit, M. N. (2010). In vitro antimicrobial activity of Muntingia calabura extracts and
fractions. African Journal of Microbiology Research, (4) 4: 304-308.