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In vivo and In vitro Phytochemical Evaluation and Quantification of Primary Metabolites from Sesamum indicum
Sharma Priyanka and Sarin Renu* Laboratory of bioactive compounds and algal biotechnology (Lab no. 9), Department of Botany, University of Rajasthan, Jaipur, Rajasthan, India. __________________________________________________________________________________
ABSTRACT Sesamum indicum Linn. is an important medicinal plant and belongs to family pedaliaceae. It is commonly known as ‘Teel’. The present investigation was aimed to investigate primary metabolites present in S. indicum. In the present study, in vivo plant parts (leaves and stem) and in vitro (calli) of S. indicum were investigated for their biochemical estimation of primary metabolites viz. total soluble sugar, starch, lipid, protein and phenol. Results showed that plant parts varied in composition of their selective metabolites. Key Words: Medicinal plant, Phytochemicals, Primary metabolites, Sesamum indicum. INTRODUCTION Traditional medicines are widespread throughout the world and comprises of those practices based on beliefs that were in existence for hundreds of years, before the development and spread of modern scientific medicines and they are still in use even today. An important feature of traditional therapy is the preference of practitioner for compound prescriptions over single substance/drug as it is being held that some constituents are effective only in the presence of others. In India, earlier the medicines used in indigenous systems of medicines were generally prepared by the practicing physicians by themselves, but now this practice has been largely replaced by the establishment of organised indigenous drug industries. The primary metabolites, in contrast, such as carbohydrate, lipid, protein and phenol are found in all plants and perform metabolic roles that are essential for plant growth and development. Primary metabolites are generally distributed within all living organism and are intimately connected with essential life processes and include ubiquitous compounds, such sugars, amino acids, or organic acid. These are involved in primary metabolic processes, such as glycolysis, respiration, or phytosynthesis. Sesamum indicum is an erect tropical annual plant growing up to 100 cm tall. It is an ancient cultigen. Today, it is mostly grown in India and the Far East (China, Korea), but its origin is probably tropic Africa although some other sources seem to favour an Indian origin. Sesame seeds have been used as a medicine since antiquity. They are considered to be antioxidant, anticancer, demulcent, emollient and laxative properties. MATERIALS AND METHODS Sesamum indicum plants were collected from Rajasthan University Campus, Jaipur from August to September. Then Specimen was deposited to the Herbarium, Department of Botany, University of Rajasthan, Jaipur for authentification (RUBL NO. 20628). For in vitro studies callus developed by tissue culture technique. The dried calli of 8 weeks old tissues were powdered and used for further bio chemical estimation. The quantitative estimation of primary metabolites was carried out using different protocols. The dried plant materials stem, leaf and callus (50 gm each) were homogenized separately in a mortar and pestle and was used for estimation of estimation of sugar and starch (Dubois et al., 1956), protein (Lowry et al., 1951), lipid (Jayaraman,1981), and Phenol (Bray and Thorpe,1954) respectively. All experiment repeated in triplicate and means (± SD) were calculated. RESULTS AND DISCUSSION The present study carried out on the plant samples revealed the presence of primary metabolites. Primary metabolites viz, total soluble sugar, starch, lipid, protein and phenol of S. indicum investigated are summarized in Table 1. In the present investigation, among all the samples higher soluble sugar, lipid and phenol level was observed in leaves of S. indicum (40.1±1.37mg/g.dw; 62.1±1.53mg/g.dw; 43.1±0.39mg/g.dw) and minimum in calli (27.8±0.13mg/g.dw; 31.0±1.64mg/g.dw; 29.7±1.43mg/g.dw). The maximum content of starch and protein was observed in stem of S. indicum (58.02±0.49 mg/gdw and 49.6±1.34 mg/gdw) while minimum
Vol. 3 (3) Jul – Sep2012
Table 1: Estimation of primary metabolites (mg/gdw) in S.8±0. the results confirm the use of the plant in traditional medicine. In similar studies carried out protein content was in leaves of M.39 S.81 Callus 27. indicum (23.8±0.0±1. 2011) .63 29.17±0.47 62. indicum Primary metabolites (mg/gdw) Plant Plant parts Sugar Starch Lipids Proteins Phenols Leaf 40.1±1.dw) (Table 1) (Fig. Trees with depleted starch reserves are more prone to die back and decline from various biotic and abiotic stressors (Gregory R. phenol etc. It may be concluded from this study that the leaves of S.1±1.8±0.45 43. protein.International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701 was in calli of S. lipid and phenol.88 58.24 49. starch. sugars. Higher content of total soluble sugar was reported in leaves of Madhuca indica (Vijayvergia and Shekhawat.13 23.43 mg/gdw = mili g per g dry weight of tissues Values represent treatment of three replicates ± SE Vol. medicinal or cosmetic (Yadav and Tyagi. Many primary metabolites lie in their impact as precursors or pharmacologically active metabolites in pharmaceutical compounds.36mg/g.37 48. Many polysaccharides purified from Chinese medicinal herbs and phenols are bioactive and possess immuno-modulating. The plant parts varied in composition of their primary metabolites.7±1.49 52.) for the normal growth and development of itself.17±0. Clerodendrum inerme and Clerodendrum phlomidis (Chahal et. 1986). The investigation can be subjected to the therapeutic uses and carry out further pharmacological evaluation. protein.8±0. 2009).0±1.com 1165 .0±0. Starch can represent 50-80% of the total nonstructural carbohydrate reserves in sugar maple.63 mg/g. Plant synthesizes primary metabolites (lipid. be it culinary.1±0. Our results were in agreement of previous reported results.64 26. starch. indicum are highly rich in lipid content.34 38. In addition. anti-tumor and antibacterial activities.indica (Vijayvergia and Shekhawat. A.5±0.ijrpbsonline. 2006).53 36.02±0.1).al.dw and 26.3±1.. Most of the active principles are found in alcoholic and aqueous extracts.36 31. The quantitative estimation of the primary metabolites yields of chemical constituents of the plants studies showed that the leaves and stems were rich in soluble sugar. 2009) and plant parts of two species of verbenaceae family viz. Plant lipid have developed products that work with diverse requirements. indicum Stem 39. 3 (3) Jul – Sep2012 www.6±1. Preliminary phytochemical screening of plant is very useful for determination of the active constituents in different solvents and their yields.
P. Department of Botany. J. Soc. Farr. Vijayavergia. R. Biol. Rosebrough. Vol. 3 (3) Jul – Sep2012 www. Dubois.com 1166 . Biochemical estimation of selective metabolites of two plants of verbenaceae family. Chahal. P. Meth. Protein measurement with the Folin-Phenol Reagent.G. and Tyagi.. Lowry. Anal. Indian bot. R. 1954. J. Yadav. O. University of Rajasthan. 4. and Thorpe. J. 2009.V. Analysis of Phenolic compounds of intrest in metabolism. 3.K. N. Chem. 193.. 7. R. 90 (3&4) : 302-305. and Shekhawat.J. Biochem. M. 2011. and Smith.. 2. R. Hamilton. J.H. Nature 168.. 1951. Gregory. New Delhi : Wiley Eastern Limited. Gilles. REFERENCES 1. 1017. 1981: Laboratory Manual in Biochemistry.. 1: 27-52. A. 1986.International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701 ACKNOWLEGEMENT The authors would like to acknowledge with thanks to Head. 265-275. starch reserves and sap sugar concentration in sugar maple.J. A colorimetric method for the determination of sugar. Canadian Journal of Forest Research.1Discovery publishing house-New Delhi. Sarin. 6.. 2(3): 203-206. 1951.. F. 8.K. A. 167. M. Jayaraman... N. H. W. The Bioscan.K.L.. Lipid Biotechnology. R. Jaipur for providing all the facilities in the department to carry out the present research work. J. Rebers.ijrpbsonline.A. Bray. 5. 2006. R. And Malwal. Vol.. and Randall. Biochemical estimation of primary metabolites of Madhuca indica GMEL. Timing of defoliation and its effect on bud development.