Preliminary phytochemical screening test of Garcinia griffithii
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Innova Ciencia Vol 4, No. 4;Apr 2012 Preliminary phytochemical screening test of Garcinia griffithii Plant Tara Kamal1, Ahmad Muzammil1, Raji Akintunde Abdullateef1, 2Muayad S. Rahma and Muhammad Nor Omar1# 1 Department of Biotechnology, Kulliyyah of Science, International Islamic University Malaysia (IIUM), Jalaln Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia. 2 Kulliyyah of Dentistry International Islamic University Malaysia 1# IIUM Research and Innovation Office, Office of Campus Director, International Islamic University Malaysia (IIUM), Jalaln Sultan Ahmad Shah, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia. E-mail: tarakamal111@hotmail.com, tarajalal@ymail.com ABSTRACT Nowadays, researchers are focusing their attention towards folk medicine for developing better drugs against microbial infections. The present study aimed to evaluate the phytochemical constituents from Garcinia griffithii plant. The plant materials were extracted using solvents with different polarities (methanol, ethyl acetate and hexane). The solvent extracts were screened and the results of screening test for phytochemicals showed that, the presence of various metabolites in different solvents. only the phenolic groups were presented in hexane and ethyl acetate extracts, whereas, The methanol extract contains terpenoids, alkaloids, flavonoids, saponins and glycosides, and those chemical compounds possess antibacterial activities against pathogenic microorganisms. Key words: G. griffithii, phytochemicals, antibacterial activities, medicinal plants. Introduction: Use of traditional medicine is an old means of curing diseases and infections. Various plants have been used in different parts of the world to treat human diseases and infections (Caceres et al., 1991; Nweze et al., 2004; Vineela and Elizabeth, 2005). The following plant, Garcinia griffithii T. Anders (Guttiferae). The family of Guttiferae were reported to be rich in natural chemical substances (Kosela et al., 1999). It is a medium-sized tree, widely distributed throughout Peninsular Malaysia. The species either grows wild or is cultivated on a small scale by villagers for its acidic fruits, much used for medicinaland culinary purposes (Permana et al., 2001). Plants in the genus Garcinia is a rich source of bioactive molecules including xanthones, flavonoids, benzophenones, lactones and phenolic acids (Patil, 2005). As part of an ongoing effort to search for phytochemical substances from Garcinia plants, it was found out that the methanol extract contains terpenoids, alkaloids, flavonoids, saponins and glycosides, crude methanol, whereas, only the phenolic groups were presented in hexane and ethyl acetate extracts. Previous phytochemical investigation on the leaves reported the presence of cytotoxic bixanthone, griffipavixanthone, which is common to G. griffithii (Xu et al., 1998) 68 IC@SCIENCERECORD.COM
Innova Ciencia Vol 4, No. 4;Apr 2012 2. Materials and Methods Collection, preparation of plant materials: The plant was collected from Taman Pertanian Sultan Haji Ahmad Shah, Kuantan, Pahang, the plant was prepared for extraction by washing off all dirt and soil residues. They were left to dry in the oven at 40oC for 72h. After drying, powdered (650g) leaf parts of G. griffithii was sequentially extracted with different solvents polarities, i.e n-hexane, ethyl acetate and methanol using Soxhlet apparatus according to Green, (2004). Each solvent was removed in vacuo by rotary evaporator at 60˚C to obtain the extracts. Then, the extracts were stored at 4˚C until further analyses. 2.2 Qualitative Phytochemical Screening of G. griffithii Extracts The qualitative chemical screening were carried out using standard procedures to identify the presence of alkaloids, tannins, flavonoids, steroids, saponins, reducing sugars and cardiac glycoside (Harborne et al., 1998). 3.2.1 Test for Flavonoids For 1 ml of each extract, a few drops of 2 % lead acetate solution were added. The configuration of yellow or orange colour was taken as evidence for the presence of the flavonoids in the plant extract. 3.2.2 Test for Alkaloids About 2 ml of the plant extract was mixed with 2ml of 2% HCl on a water bath. A few drops of Drangendorff’s reagent were then added to 1 ml of the filtrate. A yellow precipitation was taken as an indication of alkaloids in the plant. 3.2.3 Test for saponins 2 ml of each plant extract with 2 ml of distilled water were added to the test tube and strongly shaken. The subsequent formation of foam after 15 – 20 minutes was a preliminary indication of the presence of saponins. 3.2.4 Test for steroids Salkowski’s test: 2 ml of CHCl3 was added to 2ml of each extract and filtered. Then, 2ml of concentrated H2SO4 was added to the filtrate. Appearance of a reddish brown layer was the indication of the steroids compound in the plant extracts. 3.2.5 Test for terpenoids Salkowski’s test: 2ml of the CHCl3 was added to the alcoholic solution of each extract and then evaporated to dryness. Then 2ml of concentrated H2SO4 was added and heated for 2 minutes. A greyish colour was taken as a positive test for the presence of terpenoids. 3.2.6 Test for tannins To 2ml of aqueous solution of each extract, 4 drops of ferric chloride were added. Appearance of green precipitation was taken as a preliminary test for the tannins compound in the plant. 69 IC@SCIENCERECORD.COM
Innova Ciencia Vol 4, No. 4;Apr 2012 3.2.7 Test of phenolic groups About 1ml of 1% gelatine reagent containing 10% of sodium chloride was added to 1ml of each filtered aqueous extract. The formation of white precipitate was predicted as the presence of the phenols. 3.2.8 Test for glycoside Keller-Killani’s Test: About 2ml of each extract were treated with 2ml of glacial acetic acid solution which was contained in 1 drop of ferric chloride solution and 1ml of concentrated H2SO4. A reddish brown colour formed at the junction of the two layers was taken as an indication of the presence of glycoside in the plant extract. RESULTS The result of the phytochemical screening of G. griffithii extracts is presented in Table 1, which revealed that the presence of various metabolites in different solvents. The methanol extract contains terpenoids, alkaloids, flavonoids, saponins and glycosides, and those chemical compounds possess antibacterial activities against pathogens according to Nweze et al., (2007), whereas, only the phenolic groups were presented in hexane and ethyl acetate etracts. The presence of tannins in G. griffithii was observed when formation of a green precipitation was formed after the addition of ferric chloride solution. There was a development of greyish colouration which is a positive reaction for the presence of terpenoids. Flavonoids were present when a yellow colouration formed after treating the sample with lead acetate. A reddish-brown layer was observed and taken as a significant result of the presence of steroids. The formation of steady foam after shaking each extract with distilled water was a positive test for the presence of saponins in the plant extracts. The appearance of a white precipitate indicated the presence of phenolic groups after the addition of gelatine reagent. A reddish brown colour was formed at the correlation of the two layers indicated the positive result of the presence of glycoside in the plant and that is a glycone portion of glycoside. 70 IC@SCIENCERECORD.COM
Innova Ciencia Vol 4, No. 4;Apr 2012 DISCUSSION The result of the phytochemical screening of G. griffithii is presented in Table 1. Which revealed the presence of various metabolites in different solvents. The methanol extract contains terpenoids, flavonoids, saponins and glycosides, and those chemical compounds possessed antibacterial activities against pathogens according to Nwaogu et al., (2007). Only the phenolic groups were presented in hexane and ethyl acetate extracts of the plant. Phenolic groups are known to exhibit medicinal as well as physiological activity (Sofowora, 1993). Flavonoids which are part of the phytochemical constituents of methanolic extract of G. giriffithii exhibit a wide range of biological activities towards hydroxyl radicals, and superoxide anion radicals, and thus promoting health in action (Ferguson, 2001). Flavonoids also exhibit anti-inflammatory, antiangionic, anti-allergic effects, analgesic and antioxidant properties (Hodek et al., 2002). Besides that, flavonoids have also been reported as anticancer, antiviral and antibacterial agents (Silva et al., 1996; Iwu, 1999; Scheck et al., 2006). These observations support the usefulness of G. griffithii in folklore remedies for the treatment of various infections. Another finding showed that the methanol extract contains high amount of saponins and polyphenols which were reported earlier to have some antimicrobial activities (Nabila et al., 2008). Tannins have remarkable activity in cancer prevention and anticancer (Li et al., 2003). CONCLUSION The present study showed that crude extracts of the leaves of Garcinia grifithii, can be used in herbal medicine. The presence of saponins in large percentages confirmed its usage by traditional healers in treating infected wounds and ulcers and also as anti toxic properties. Flavonoids are known to be anti-inflammtory, antiallergic, antiviral and antibacterial agents. Tannins has remarkable activity in cancer prevention and anticancer. Thus, G. griffithii support the traditional medicinal use in the treatment of ailments caused by microorganisms. 71 IC@SCIENCERECORD.COM
Innova Ciencia Vol 4, No. 4;Apr 2012 REFERENCE Caceres, A. L., Lopez, B. R., Giron, M. A., & Logemann, H. (1991). Plants used in Guatemala for the treatment of dermatophytic infection. I. Screening for antimycotic activity of 44 plant extracts. J. Ethnopharmocol, 31(263-276). Ferguson, L. R. (2001). Role of plant polyphenols in genomic stability. Mutat. Res., 475, 9-111. Green, R. J. (2004). Antioxidant Activity of Peanut Plant Tissues. North Carolina State University, USA. Harborne, J. B. (1998). Phytochemical Methods. Chapman and Hall Ltd London, 49-188. Hodek, P., &Trefil, P. (2002). Flavonoids-Potent and versatile biologically active compounds interacting with cytochrome. Chemico-Biol. Int., 139(1), 1-12. Iwu, M. (1999). Garcinia cola: A new adaptogen with remarkable immunostimulant, antiinfective and anti-inflammatory properties. A Colloquium on Garcinia kola presented in International Conference on Ethnomedicine and drug discovery. Maryland, USA, 26. Li, H., Wang, Z., & liu, Y. (2003). Review in the studies on tannins activity of cancer prevention and anticancer Zhong-Yao-Cai, 26(6), 444-448. Kosela, S., Hu L., H., Yip S. C., Rachmatiah, T., Sukri, T., Daulay, T. S., Tan, G. K., Vital, J. J., & Sim, K. Y. (1999). Dulxanthone E: a pyranoxanthone from the leaves of Garcinia dulcis. Phytochemistry, 52, 1375. Nabila, B., et al. (2008). Antioxidant and antimicrobial activities of the Pistacia lentiscu and Pistacia atlantica extracts. Afri.j.pharm.pharmacol, 2(2), 22-28. Nwaogu, L.A., Alisi C.S., Ibegbulem C.O., & Igwe, C.U. (2007). Phytochemical and antimicrobial activity of ethanolic extract of Landolphia owariensis leaf. African Journal of Biotechnology, 6, 890-893. Nweze, E. L., Okafor, J. I., & Njokn, O. (2004). Antimicrobial activities of methanolic extracts of Trema guinensis (Schumm and Thorn) and Morinda Lucida Benth used in Nigeria. Bio. Res., 2, 39-46. Patil, B. P. (2005). Kokum, Brochure, Western Ghats Kokum Foundation, Goa. Permana, D., Lajis N.H, Mackeen M.M, Ali A.M, Aimi N., Kitajima M. (2001). Isolation and bioactivities of constitutents of the roots of Garcinia atroviridis. . J Nat Prod. , 64(7), 976-980. 72 IC@SCIENCERECORD.COM
Innova Ciencia Vol 4, No. 4;Apr 2012 Scheck, A. C., Perry, K., Hank, N. C., & Clark, W. D. (2006). Anticancer activity of extracts derived from the mature roots of Scutellaria baicalensis on human malignant brain tumor cells. Biomed Centr. Complement. Alternat. Med., 6, 27. Silva, O., Duarte, A., Cabrita, J., Pimentel, M. D. A., & Gomez, E. (1996). Antimicrobial activity of Guinea-Bissau traditional remedies. J. Ethnopharmacol, 50, 53-59. Sofowora, A. (1993). Medicinal and Traditional medicine in Africa. (2ed.). Ibadan, Nigeria. p.130: published by Spectrum Books Limited Spectrum Books Limited. Vineela, C. H., & Elizabeth, K. M. (2005). Antimicrobial activity of marine algae of Visakhapatnam city, Andhra Pradesh. Asian J. Microbiol. Biotech.Environ. Sci, 7, 209- 212. Xu, Y. J., Cao, S. G., & Wu, X. H. (1998). Griffipavixanthone,a novel cytotoxic bixanthone from Garcinia griffithii and pavifolia. Tetrahedron Lett., 39, 9103-9106. 73 IC@SCIENCERECORD.COM
Innova Ciencia Vol 4, No. 4;Apr 2012 Table 1. Phytochemical screening of G. griffithii (leaves) Phytochemical Hexane Ethylacetate Methanol components extract extract extract Phenolic + + + group flavonoids _ _ ++ alkaloids _ _ + tannins _ _ + Saponins _ _ +++ Terpenoids _ _ ++ Glycoside _ _ ++ Steroids _ + + + = Trace, ++ = Moderate, +++ = Abundant, = Absent 74 IC@SCIENCERECORD.COM
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