Krishnamurthy S.R et al. / Journal of Pharmacy Research 2011,4(6),1610-1613 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Phytochemical screening of of umbellatum Burm.: A medicinal of Central Western Ghats *Krishnamurthy S.R. and Asha B. Department of Applied Botany, Kuvempu University, Shankaraghatta – 577 451, Shimoga District, Karnataka, INDIA Received on: 11-02-2011; Revised on: 16-03-2011; Accepted on:21-04-2011 ABSTRACT Memecylon umbellatum Burm., a medicinal plant belongs to the family Melastomaceae. The plant is used in the treatment of diabetes, herpes, gonorrhea, leucorrhea and skin diseases. The young and mature leaves were collected form semi evergreen forests of central Western Ghats. The hot and cold extracts of leaves in different solvents namely petroleum ether, chloroform, ethanol and aqueous, were subjected for phytochemical screening for carbohydrates, proteins, tannins, saponins, terpinoids, flavonoids, steroids, glycosides, alkaloids, phenols and lignin. The glycosides and lignin were absent in all the extracts of young and mature leaves. The quantitative analysis of phenols, tannins, steroids, alkaloids, flavonoids, lignins, proteins and carbohydrates reveal that alkaloid and lignin were absent in both young and mature leaves whereas phenols, tannins, flavonoids and steroids were recorded and their variation between young and mature leaves were narrow.

Key words: Central Western Ghats, medicinal , Melastomaceae, Memecylon umbellatum, semi ever green forests. INTRODUCTION The Memecylon belongs to the family Melastomaceae. Sivu et al. [1] mentioned 300 from 2 species of Memecylon. Agarwal and Rastogi [5], isolated umbelactone (4- species of Memecylon which are found in wide range of habitats from deciduous, semi hydroxymethyl-3-methyl-but-2-ene-4,1-olide), a new constituent of M. umbellatum. Joshi evergreen, evergreen and montane shoals with a wide range of altitude from sea level to et al. [6] isolated fatty acids like octocosonoic acid, cerotic acid, ethyl palmitate, palmitic 2500 mts. Memecylon umbellatum Burm. is a small handsome tree, it is locally called as acid and butyric acid from the n-hexane extract of the roots. Amalraj and Ignacimuthu [7] Adachare and the English name is iron-wood tree. All the parts of the plant is used as evaluated the hypoglycaemic effect of alcoholic extract of the leaves in normal and alloxan medicine, the fruits are edible, are eaten in time of famine and they are quite safe. The diabetic mice. Krishnamurthy and Asha [8] determined proximates, nutritive value and leaves are used in the dyeing industry to dye wool, silk and grass mats [2]. Ram Rastogi elemental composition of M. umbellatum from the two distinct climatic regions of Karnataka and Mehrotra [3] isolated and characterized the umbellactone-b-amyrin, sitosterol, its from the young and mature leaves and reported that young leaves are more nutritive. glycoside, olenalic and ursolic acids. Lowry [4] reported the occurrence of Anthocyanins Puratchikody and Nagalakshmi [9] evaluated the wound healing activity of alcoholic extract of leaves in rats in the form of ointment. The leaves have been being used for the

INDIA preparation of herbal formulations. The two major tribal groups of south Tamil Nadu, India, used the shade dried powder with cup of water and boiled rice and kept it for overnight. They take one teaspoon of this orally in the morning for 40 days or until cure to treat diabetes [10]. The paste of the leaves with a piece of roof tile is applied on herpes infected parts [11]. The infusion of leaf is used against snake- bite [12]. In the present study, an attempt is made to screen both the hot and cold extracts of young and mature leaves of Memecylon umbellatum Burm. for its phytochemicals. KARNATAKA MATERIALS AND METHODS

Study Area The samples were collected at semi ever green forests of Koppa in Chikmagalore districts of Karnataka, India. It is geographically situated on 13o 33’ N latitude and 75o 21’ 59" E longitude. The altitude is 2506 feet MSL. The temperature varies from 21.1oC to 28.2oC. It receives 1925 mm annual rainfall (Map 1).

Identification and description Having collected and brought to the laboratory, the specimen was identified. The plant is an erect shrub or a small tree, 2-3 mts tall and branches glabrous, leaves petiolated, flowers on pedicilate, pedunculate cyme, cymes branched lateral, the ultimate branches are umbellulate. Leaves ovate or ovate to elliptic, 2 inches long and 0.75 inches broad, emarginated apex, yellowish when dry, nerves not visible, calyx companulate and the disc rays are conspicuous [13]. (Specimen voucher No. KU/AB/BA-4).

* Preparation of Samples The collected leaves were washed in the running tap water and alcohol to remove dust particles from the surface. The leaves were shade dried and powdered in electric blender. The powdered samples were packed in air-tight plastic containers and stored in the refrigerator for further analysis.

* Koppa Preparation of Extracts The powdered material was subjected to both hot and cold extraction.

Hot Extraction A known amount of shade dried powder of leaf was filled in the thimble and extracted successively with various solvents of gradual increasing polarities from petroleum ether to *Corresponding author. distilled water through chloroform and ethanol using a Soxhlet extractor for 48 hrs. All the extracts were concentrated using rotary flash evaporator. After complete solvent evaporation, Krishnamurthy S.R. the extract was preserved at 5 o C in airtight bottles until further use [14,15]. Department of Applied Botany, Kuvempu University, Cold Extraction Shankaraghatta – 577 451, Powdered samples was weighed and soaked in different solvents for 24 hrs with intermittent Shimoga District, Karnataka, INDIA shaking. The plant extract was then collected and filtered through Whatman No.1 filter

Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1610-1613 Krishnamurthy S.R et al. / Journal of Pharmacy Research 2011,4(6),1610-1613 paper. The extract was concentrated using a rotary evaporator, then preserved in air tight Saponins bottles at 4o C until further use [16,17]. Hot and cold ethanol extracts of both young and mature leaves showed +ve response for both foam and haemolysis test. Preliminary screening of primary and secondary metabolites The extracts obtained from both hot and cold extractions were subjected for preliminary Triterpenoids phytochemical screening of primary and secondary metabolites [Carbohydrates (Molisch’s, Only hot and cold ethanol extract of both hot and cold extract of young and mature leaves Benidict’s and Fehling’s tests), Proteins (Million’s, Xanthoprotic, Biuret and Ninhydrin showed +ve response to Salkowaski test. tests), Tannins (Fecl and Gelatin tests), Saponins (Foam and Haemolysis tests), 3 Flavonoids Triterpenoids (Salkowaski test), Flavoniods (Fecl3, Shinoda, NaoH, Lead acetate, Mineral acid and Zinc hydrochloric tests), Steroids (Salkowaski test), Glycosides (Killer Killani Hot and cold ethanol extracts of both samples showed +ve response to FeCl3, Shinoda and test), Alkaloids (Mayer’s, Wagner’s, Sonneuschinis and Tannic acid tests), Phenols, zinc hydrochloric tests. Only hot and cold aqueous extracts of both young and mature (Phenol and Ellagic tests), Lignins (Labat and Furfuraldehyde tests)] [18,19]. leaves showed +ve response to mineral acid test. Hot ethanol extract of mature leaves, cold aqueous extract of young leaves and chloroform and aqueous extracts of mature leaves Quantitative estimation of primary and secondary metabolites showed +ve response to lead acetate test. Hot petroleum ether and chloroform extracts of The powdered plant material was subjected for Quantitative analysis of secondary young leaves, chloroform extracts of mature leaves and cold aqueous extract of young metabolites. (Phenols [20,21], tannins [22], steroids [23], alkaloids [24], flavonoids [25], proteins leaves, chloroform and aqueous extract of mature leaves showed +ve response to NaOH [26] and carbohydrates [27] ). test.

RESULTS Steroids Preliminary screening of primary and secondary metabolites Salkowaski test showed +ve to only ethanol extracts of both hot and cold young and The presence or absence of phytochemicals for different tests are given in Table 1. mature leaves. Table 1. Screening of phytochemicals of M. umbellatum

Hot extract Cold extract Young leaves Mature leaves Young leaves Mature leaves Carbohydrates PE Chl Eth Aq PE Chl Eth Aq PE Chl Eth Aq PE Chl Eth Aq

Molisch’s test + + + + + + + + + + + + + + + + Benidict’s test + - + ------+ - - - - - Fehling’s test - - + - - - + - - - + - - - + - Proteins Million’s test + + - + - - - + - - + + - + - + Xanthoprotic test + + - + ------Biuret test ------+ ------Ninhydrin test ------Tannins

Fecl3 test - - + - - - + - - - + - - - + - Gelatin test ------Saponins Foam test - - + - - - + - - - + - - - + - Haemolysis test - - + - - - + - - - + - - - + - Triterpenoids

Salkowaski test - - + - - - + - - - + - - - + - Flavoniods

Fecl3 test - - + - - - + - - - + - - - + - Shinoda test - - + - - - + - - - + - - - + - NaoH test + + - - - + - - - - - + - + - + Lead acetate test ------+ - - - + + - - + - Mineral acid test - - - + - - - + - - - + - - - + Zinc hydrochloric test - - + - - - + - - - + - - - + - Steroids Salkowaski test - - + - - - + - - - + - - - + - Glycosides Killer killani test ------Alkaloids Mayer’s test ------Wagner’s test ------Sonneuschinis test ------Tannic acid test ------Phenols Phenol test - - + - - - + - - - + - - - + - Ellagic test ------Lignins Labat test ------Furfuraldehyde test ------PE=Petroleum Ether;Eth=EthanolChl=Chloroform;Aq=Aqueous ;‘+’ = Positive;‘-’ = Negative

Carbohydrates Glycosides Both hot and cold extracts of young and mature leaves showed +ve response to Molisch’s All cold and hot extracts of young and mature showed –ve response to Killer Killani test. test. Hot extract of petroleum ether and ethanol of young leaves and cold of ethanol extract of young leaves showed +ve response to Benidict’s test. Both hot and cold ethanol Alkaloids extracts of young and mature leaves showed +ve response for Fehling’s test. Only hot aqueous extract of young leaves and cold ethanol extract of young leaves showed negative response to Sonneuschinis test. Mayer’s, Wagner’s and tannic acid tests showed Proteins –ve result to both hot and cold extracts of young and mature leaves. Hot petroleum, chloroform and aqueous extracts of young leaves and aqueous extract of mature leaves showed +ve response to Million’s test and cold ethanol and aqueous Phenols extracts of young leaves and chloroform and aqueous extracts of mature leaves showed +ve Phenol test showed +ve response to only hot and cold ethanol extracts of young and response to Million’s test. Only hot petroleum ether, chloroform and aqueous extracts of mature leaves and all the extracts showed –ve response to Ellagic test. young leaves showed +ve for Xanthoprotic test. Only hot ethanol extract of mature leaves showed +ve for Biuret test and all the extracts of both the samples showed –ve for Lignins ninhydrin test. Both Labat and furfuraldehyde tests showed –ve response to all extracts of young and mature leaves. Tannins Quantitative estimation of primary and secondary metabolites Both hot and cold ethanol extract of young and mature leaves showed +ve result for FeCl3 test and Gelatin test showed –ve response for both hot and cold extracts of young and The values of protein ranges between 2.85 to 3 mg/g in mature leaves and 1.75 to 1.9 mg/ mature leaves. g in young leaves with an average of 2.95 mg/g and 1.85 mg/g in mature and young leaves respectively. The young leaves contain an average of 10.7 mg/g and mature leaves contain Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1610-1613 Krishnamurthy S.R et al. / Journal of Pharmacy Research 2011,4(6),1610-1613

30 The presence of some of these phytochemicals has been demonstrated previously by other Mature researchers. For example, the percentage of tannins in different parts of M. umbellatum has [34] 25 Young been estimated by Killedar and More . They estimated that the highest percentage of tannin content was found in bark than in leaves, roots and stem bark and inflorescence [5] 20 showed minimum amount of tannins. Agarwal and Rastogi isolated and characterized umbelactone a new compound from M. umbellatum. Joshi et al.[6] determined fatty acids in n-hexane extract of the roots of M. umbellatum. Lowary [4] found acylated anthocyanins 15 from two species of Memecylon. The presence of phenolic compounds in the leaves mg/gm indicates that this plant may be used as antimicrobial agent because phenols and phenolic 10 compounds have been extensively used in disinfections [35]. The aromatic compounds with hydroxyl group are widespread in the plant kingdom. They occur in all parts of the 5 plants phenols are said to offer resistance to diseases and pests in plants [36]. The presence of phenol further indicated that the leaves of M. umbellatum may act as anti-inflammatory, 0 anti-clotting, anti-oxidant, immune enhancers and hormone modulator. The extensive [37, 38] Lignins

Phenols Tannins research is going on, on the phenols as they are disease preventives . Phenols have Steroids Proteins Alkaloids

Flavonoids been responsible in having the ability to block specific enzymes which causes inflammation.

Ca rbohydrates They also play an important role in modifying the prostaglandin pathways and thereby Plant metabolites they protect platelets from clumping [39].

Fig. 1. Quantitative estimation of secondary metabolites Tannins are complex moieties produced by majority of plants as protective substances, 7.33 mg/g of carbohydrates and the average values ranged between 9.8 to 11.6 mg/g and they have wide pharmacological activities. They have been used since past as tanning 6.8 to 8 mg/g in young and mature leaves respectively. The values of phenols ranged agents and they posses astringent, anti-inflammatory, antidiarrhoeal, antioxidant and [34] between 24.6 to 26.4 mg/g and 22.8 to 24.6 mg/g with an average of 25.4 and 23.6 mg/ antimicrobial activities . Tannins have astringent properties, they heal wounds and [40] g in young and mature leaves respectively. The values of tannins ranged between 6.2 to inframed mucous membrane . They are also known as antimicrobial agents. They are 6.4 mg/g with an average of 6.26 and between 5.5 to 5.1 mg/g with an average of 5.6 mg/ water soluble polyphenols that are present in many plant foods and precipitate proteins. g in mature and young leaves respectively. The young leaves contain high values of They also prevent the development of microorganisms by precipitating microbial protein [36] steroids than mature leaves and the values ranged between 13.6 to 25.2 mg/g 5.8 to 13.6 and making nutritional proteins unavailable for them . The tannins also have various mg/g with an average of 20.2 in young leaves and 9.33 mg/g in mature leaves. The physiological effects like anti-irritant, antisecretolytic, antiphlogistic, antiparasitic effects [41] average values of flavonoid is 3.44 mg/g in young leaves and 1.83 mg/g in mature leaves . The presence of tannins in M. umbellatum strongly supports that it may be used in and the values ranged between 2.56 to 4.06 mg/g and 1.74 to 1.92 mg/g in young and treating wounds, varicose ulcers, hemorrhoids, frostbite and burns in herbal [42, 43] mature leaves respectively. The alkoloids and lignins were absent in both young and medicine . mature samples. (Table2). Between the two leaf samples mature leaf recorded high values of phenols, tannins and proteins where as young leaves recorded high values of steroids, Flavonoids are a group of polyphenolic compounds which influence the radical scavenging, flavonoids and carbohydrates. The alkaloids and lignin were absent not only in young inhibition of hydrolytic and oxidative enzymes and also act as anti-inflammatory agent [44] leaves but also in mature leaves. . The flavonoids show antioxidant activity and their effects on human nutrition and health is considerable. The mechanisms of action of flavonoids are through scavenging or Table 2. Minimum, maximum and average values of chelating process [45,46]. They also inhibit microbes which are resistant to antibiotics [47]. DISCUSSION primary and secondary metabolites in mg/gm (pro- The presence of flavonoids in M. umbellatum reveals that the plant is used for the Plant foods contain constituents teins, carbohydrates, phenol, tannins, steroids, fla- management of cardiovascular diseases and oxidative stress since flavonoids are biologic such as flavonoids, saponins, vonoids, lignins, and alkaloids) of M. umbellatum antioxidants. Flavonoid may help in providing protection against some diseases such as tannins phenolics etc, which have Secondary Metabolites Mature Young oxidative stress, cellular damage [48]. Oxidative stresses have been linked to cancer, been assessed for their anti-oxidant, ageing, atherosclerosis, inflammation, ischemic injury and neuro degenerative diseases anti-mutagenic, anti carcinogenic Proteins 2.95 1.85 (1) [49] [28] (2.85-3.00) (1.75-1.90) . and other biological effects . The Carbohydrates 7.33 10.7 medicinal properties of plants can (6.80-8.00) (9.80-11.60) Saponins are a special class of glycosides which have soapy characteristics [50], they have be assessed by their response to Phenols 25.4 23.6 active antifungal properties [36]. The presence of saponins in M. umbellatum contribute to attacks from insect predators and (24.6-26.4) (22.8-24.6) Tannins 6.26 5.6 its medicinal value. Saponins inhibit Na+ efflux by the lockage of the entrance of the disease organisms. This is achieved (6.2-6.4) (5.5-5.7) Na+out of the cell. This leads to higher Na+ concentration in the cells, activating a Na+ - by the accumulation of Steroids 9.33 20.2 Ca2+ anti porter in cardiac muscle. The increase in Ca2+ in flux through this antiporter, phytochemicals at the sites of (5.8-13.6) (13.6-25.2) Flavonoids 1.83 3.44 which strengthens the contractions of heart muscle [51]. infusion of plants several of which (1.74-1.92) (2.56-4.02) are insecticidal, anti-bacterial, anti- Lignins - (2) - The presence of phenols, tannins, flavonoids and sponins justify the use of leaf extracts for fungal etc. [29, 30]. Alkaloids - - the treatment of diabetes. Amalral and Ignacimuthu [7] evaluated the hypoglycaemic effect (1) = The values in parenthesis indicates the average of alcoholic extract of the leaves in normal and alloxan diabetic mice. Tiwari and Rao [52] Phytochemical analysis reveals the (2) = Not recorded said that besides antioxidant properties the phenols are also inhibit alpha- amylase, active biological components of sucrase as well as the action of sodium glucose-transporter 1 (S-GLUT-1) of the intestinal medicinal plants. The qualitative and quantitative analysis of different solvent extracts of brush border, hence results in the antidiabetic action. In addition isoflavones, tannins, M. umbellatum were carried out in both young and mature leaf samples. The result of chlorgenic acids and crude saponins are also posses potent S-GLUT-1 mediated inhibition qualitative analysis reveals that the hot and cold ethanol extracts of both young and of glucose transporter and again results in the antidiabetic activity. During our present mature leaves showed positive results for majority of the tests. The quantitative analysis study the qualitative and quantitative studies reveals that alkoloids are absent in both result reveals that the phenols, tannins, steroids, flavoniods, proteins and carbohydrates young and mature leaves. However, Maridass [53] reported the presence of alkoloids and were present in both the samples but alkaloids and lignin were not recorded in both the triterpens in the 50% ethanol extract of leaves of M. umbellatum and absence of samples (Table 1 and 2 respectively). This result shows high level of its possible flavonoids, saponins and tannins. Where as the present study reveals the presence of medicinal values. Some of these active components have been demonstrated to posses flavonoids, saponins and tannins not only in the young leaves but also in mature leaf antinutritional effects, following their ability to reduce palatability and digestibility of samples. feedstuff. Some of these constitute may be completely harmful to both man and farm [31] animals and some are species specific as observed in the case of tannins . In table 2 the The present study scientifically validates the use of this plants in traditional medicinal quantity of these phytochemical were shown. The young samples showed higher levels of plant and phytochemical data will be helpful for the standardization and quality control of steroids, flavanoids and carbohydrates and mature samples showed higher leaves of precious indigenous drug and also pharmaceutical industries. phenols, tannins and protein and both the samples were failed to record alkaloid and lignins. ACKNOWLEDGEMENT The authors wish to thank the Chairman, Department of Applied Botany, Kuvempu The protein contributes to the formation of hormones and other metabolites which are University for providing laboratory facilities and Asha, B. thank Kuvempu University for [32] required for the growth and development of plants and Mau et al. emphasized the awarding Junior Research Fellowship. contribution of low level proteins to the formation of hormones which controls a variety of body functions such as growth, repair and maintenance of body proteins. Carbohydrate REFERENCES [33] content indicates the storage and transport of metabolic fuel and energy source . [1] Sivu AR, Pradeep NS and Pnadurangan AG, The status of genus Memecylon Linn. () in Western Ghats, Souvenir and Abstracts, XX Annual Conference of Indian Association for Angiosperm Taxonomy and International Symposium on “Taxonomy, Plant Diversity Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1610-1613 Krishnamurthy S.R et al. / Journal of Pharmacy Research 2011,4(6),1610-1613 and Conservation”, held at Department of Botany, Bharathiar University, Coimbatore – 641 046, [26] Lowry OH, Rosebroug NG, Farra AL and Pandall RL, Protein measurement with folin phenol Tamil Nadu, 2010. reagent, J. Biol. Chemistry, 193, 1951, 265-275. [2] Kolkata (http://rajbhavankokata.gov.in/pdf/occassional%20paper5.pdf.) [27] Hedge JE and Hofreiter HT, In: Carbohydrate chemistry, Dhistler RL and Miller JN (eds), Acad [3] Ram P Rastogi and Mehrotra BN, Compendium of Indian Medicinal Plants. Central Drug Research Press, New York, 1962. Institute, Lucknow, 2, 1991, 1970-1979, 453. [28] Krishnaswamy K and Raghuramulu N, Bioactive phytochemicals with emphasis on dietary [4] Lowry JB, Anthocyanins of the Melastomataceae, Myrtaceae and some allied families, practices. In. J Med Res., 108, 1998, 167-181. Phytochemistry, 15, 1976, 513-516. [29] Walker RL, Functional aspects of phenolic compounds in plants. The biology of plant phenolics. 2nd [5] Agarwal SK and Rastogi RP, Umbelactone (4-hydroxymethyl-3- methyl-but-2-ene-4, 1-olide) Edn., Edward Arnold Ltd. London, 1975, 23-32. new constituent of Memecylon umbellatum, Phytochemistry, 17, 1978, 1663-1664. [30] Amen OM, Fatope OM, Usman LA and Adebayo SA, Bioactive metabolites in improved cowpea [6] Joshi H, Joshi AB, Sati H, Gururaja MP, Shetty PR, Subramanyam EVS and Satyanarayana D, seeds. Afr. J. Biotech., 4, 2005, 513-516. Fatty acids from Memecylon umbellatum (Burm.), Asian J. Research Chem., 2(2), 2009, 178-180. [31] Odebiyi OO and Sofowora EA, Phytochemical screening of Nigerian medicinal plants 2nd OAU/ [7] Amalraj T and Ignacimuthu S, Evaluation of the hypoglycaemic effect of Memecylon umbellatum STRC inter-African Symposium on traditional pharmacopoeia and African medicinal, 1979, 216- in normal and alloxan diabetic mice, Journal of Ethnopharmacology, 62, 1998, 247-250. 220. [8] Krishnamurthy SR and Asha B, Determination of chemical components of Memecylon umbellatum [32] Mau JL, Miklus MB and Beelman RB, Shelf life studies of foods and Beverages charalambous E.D. Burm.–A medicinal plant, Karnataka, India, Pakistan Journal of Nutrition, 9(5), 2010, 438-443. Chem. Biol. Phys. Nutr. Aspect, 57, 1999, 475-477. [9] Puratchikody A and Nagalakshmi G, Wound healing activity of Memecylon umbellatum Burm. [33] Alli Smith YR, Determination of Chemical composition of Senna siamea (Cassia leaves). Pakistan Journal of Plant Sciences, 2(2), 2007, 179-186. Journal of Nutrition, 8(2), 2009, 119-121. [10] Ayyanar M, Sankarasivaraman K and Ignacimuthu S, Traditional herbal medicines used for the [34] Killedar SG and More HN, Estimation of tannins in different parts of Memecylon umbellatum treatment of diabetes among two major tribal groups in South Tamil Nadu, India, Ethnobotanical Burm., Journal of Pharmacy Research, 3(3), 2010, 554-556. Leaflets, 12, 2008, 276-280. [35] Okwu DE, Evaluation of the chemical composition of indigenous spices and flavouring agents. [11] Maruthi KR, Krishna V, Manjunatha BK and Nagaraja YP, Traditional Medicinal Plants of Global J. Pure and Applied Sci., 8, 2001, 455-459. Davanagere District, Karnataka with Reference to Cure of Skin Diseases, Environment and Ecology, [36] Sadipo OA, Akanji MA, Kolawole FB and Odutuga AA, Bio Res Commun, 3, 1991, 171. 18(2), 2000, 441-446. [37] Okwu DE and Omodamiro OD, Effects of hexane extract and phytochemical content of Xylopia [12] Kshirsagar RD and Singh NP, Some less known ethnomedicinal uses from Mysore and Coorg acthiopica and Ocimum gratissium on the uterus of Guinea pig., Bio. Res., 2005, 3. districts, Karnataka state, India, Journal of Ethnopharmacology, 75, 2001, 231-238. [38] Duke J, Handbook of biological active phytochemicals and their activities, BOCA Raton (RL), CRC [13] Gamble JS, Flora of Presidency of Madras, Published under the authority of the secretary of state Press, 1992, 99-131. for India in council, London, Adlard and son, Limited, 21, Hart Street, W.O, 1, 1928, 500. [39] Okwu DE, Phytochemicals, vitamins and mineral contents of two Nigerian medicinal plants. [14] Babu S, Satish S, Mohana DC, Raghavendra MP and Raveesha KA, Antibacterial evaluation and International Journal of Molecular Medicine and Advance Sciences, 1(4), 2005, 375-381. phytochemical analysis of some Iranian medicinal plants against plant pathogenic Xanthomonas [40] Okwu DE and Okwu ME, Chemical composition of Spondias mombia Linn. Plant parts. J. Sustain pathovars, Journal of Agricultural Technology, 3(2), 2007, 307-316. Agric. Environ., 6, 2004, 140-147. [15] Srinivasan Kesavan, Devarajan Natarajan, Chokkalingam Mohanasundari, Chinthambi [41] Asquith TN and Butler LG, Phytochemistry, 25(7), 1986, 1591-1593. Venkatakrishnan and Nandakumar Nagakumarugan, Antimicrobial, preliminary phytochemical [42] Igboko DO, Phytochemcial studies on Garcinia kola Heckel. M.Sc., Thesis University of Nigeria and pharmacognostical screening on the leaves of Vicoa indica (L.) DC. Iranian Journal of Nsukka Nigeria, 1983, 202. Pharmacology and Therapeutics, 6(1), 2007, 109-113. [43] Maduinyi I, Biochemical and pharmacological studies of active principles of the seeds of Garcinia [16] Kubmarawa D, Ajoku GA, Enweram NM and Okorie DA, Preliminary phytochemical and kola Heckel. M.Sc., Thesis University of Nigeria Nsukka Nigeria, 1983, 108. antimicrobial screening of 50 medicinal plants of Nigeria. African Journal of Biotechnology, 6(14), [44] Frankel E, Nutritional benefits of flavonoids, International conference on food factors: chemistry 2007, 1690-1696. and cancer prevention, Hamamatsu, Japan, Abstracts, C6-2, 1995. [17] Karthikeyan A, Shanthy V and Nagasathaya A, Preliminary phytochemical and antibacterial [45] Kessler M. Ubeand G and Jung L, Anti and prooxidant activity of rutin and quercetin derivatives, screening of crude extract of the leaf of Adhatoda vasica L., International Journal of Green J. Pharm and Pharmacol, 55, 2003, 131-142. Pharmacy, 2009, 78-80. [46] Cook NC and Samman S, flavonoids chemistry, metabolism, cardioprotective effects and dietary [18] Harborne JB, Phytochemical Methods. 3rd Ed, Chapman and Hall, Madras, 1998. sources. Nutritional Biochemistry, 7, 1996, 66-76. [19] Evans WC, Pharmacognosy, 13th Ed, Barllier – Tindall; London, 1989. [47] Linuma M, Tsuchiya H, Salo M, Yokoyama J, Ohyama M, Ohkawa Y et al., J. Pharmacol, 46(11), [20] Fiegel F, Spot tests in organic analysis. Elsevier publications Co., London, 1960, 143. 1994, 892-895. [21] Gibbs RD, Chemotaxonomy of flowering plants. MC grill Queens university press, Montreal, 1974, [48] Burlon GW and Ingoid, KU, B-Carotene, an unusual type of lipid antioxidant. J. Sci., 224, 1984, 573. 523-619. [49] Palozza P, Pro-oxidant actions of Carotenoids in Biologic Systems. Nutr. Revolution, 56, 1998, 257- [22] Folin D and Denis W, A calorimetric estimation of phenols (phenol derivatives) in Urine, J. Biol, 266. Chem., 22, 1939, 305-308. [50] Fluck H, Medicinal plants and their uses, W. Feulshom and comp. Ltd, New York, 1973, 7-15. [23] Sanchez GL, Medina Acevedo and Solo RR, Spectrophotometric determination of disogenin in [51] Schneider G and Woliling J, Synthetic Cardenolides and related compounds. Current Organic Dioscores composite following thin layer chromatography. Analyst, 97, 1972, 973. Chemistry, 2004, 8. [24] Ikan R, Natural products : A laboratory guide. Academic press, London, 1969, 178-260. [52] Tiwari AK and Rao JM, Diabetic mellitus and multiple therapeutic approaches of phytochemicals: [25] Swain T and Shillif WE, The phenolic constituents of Brunus domestica, the qualitative analysis of present status and future prospects. Curr, Sci, 83(1), 2002, 30-37. phenol constituent, S.J. Sci. Food. Agric., 10, 1959, 63-68. [53] Maridass M, Survey of phytochemical diversity of wild plants in Tirunelveli Hills, Western Ghats, South India. International Journal of Advances in Pharmaceutical Sciences, 1, 2010, 128-132.

Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.4.Issue 6. June 2011 1610-1613