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Preliminary Phytochemical and Pharmacognostic Analysis of Bauhinia Tomentosa

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Preliminary Phytochemical and Pharmacognostic Analysis of Bauhinia Tomentosa Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 2015, 7(4):271-277 ISSN : 0975-7384 Research Article CODEN(USA) : JCPRC5 Preliminary phytochemical and pharmacognostic analysis of Bauhinia tomentosa R. Balabhaskar # and K. Vijayalakshmi* #Bharathiyar University, Tamil Nadu, India *Dept. of Biochemistry, Bharathi Women’s College (Autonomous), Chennai, Tamil Nadu, India _____________________________________________________________________________________________ ABSTRACT Bauhinia tomentosa is one of the best, versatile and most commonly used house hold remedy for many manifestations. The present study is aimed to investigate the pharmacognostic analysis of the leaves and the phytochemicals present. The leaves were fixed in TBA, sectioned and then stained with toluidine blue. The leaves were dried and extracted with petroleum ether, ethyl acetate, ethanol and water. Phytochemical analysis was done with all four extracts using standard procedures. The pharmacognostical studies revealed the presence of calcium oxalate crystals and different cell components were studied and measured. The extractive yield was found to be high in aqueous and ethanol extracts. Most of the phytochemicals were found to be present in the ethanol extract. Further studies are to be carried out to assay the bioactivity of the phytochemicals and the medicinal values of the various phytochemicals present in Bauhinia tomentosa. Key words : Bauhinia tomentosa , pharmacognosy phytochemicals, ethanol extract _____________________________________________________________________________________________ INTRODUCTION Medicinal plants are part and parcel of human society to combat diseases from the dawn of civilization [1].There is a widespread belief that the herbal medicines are healthier and safer than synthetic ones [2].There exists a plethora of knowledge, information and benefits of herbal drugs in our ancient literature of Ayurveda, Siddha, Unani and Chinese medicine. Herbal medicines are in great demand in the developed as well as developing countries for primary health care because of their wide biological and medicinal activities, higher safety margins and lesser costs [3, 4]. Bauhinia is a genus of more than 200 species of flowering plants in the sub family Cesalpinioidae of the large flowering plant family, Fabacae with a tropical distribution. Bauhinia tomentosa commonly known as Yellow bell orchid tree belongs to Fabaceae family is one of the best, versatile and most commonly used household remedy for many manifestations [5]. The generic name commemorates the bauhin brothers Jean and Gaspard, the swiss botanists; the two lobes of the leaf exemplify the two brothers. Tomentosa derived from tomentose, meaning with dense, interwoven hairs. It is commonly known as ‘Kanchini’ in Tamil and ‘Phalgu’ in Sanskrit [6]. It is usually a scrambling, many stemmed shrub or small tree reaching 4m (max.8) in height, the branches often drooping, with many slender twigs [7].Leaves are deeply divided for almost half their length, with a small apical appendage between the lobes; each lobe is oval to almost elliptic, most often small about 2.5 x 2.5 cm, but may be up to 8cm, the colour of the leaf is pale fresh green. The apex of each lobe is broadly tapered. The base of the whole leaf is shallowly lobed and the length of the leaf stalk is usually around 10-30 mm long. [8]. 271 R. Balabhaskar and K. Vijayalakshmi J. Chem. Pharm. Res., 2015, 7(4):271-277 ______________________________________________________________________________ The dried leaf, flower bud and a decoction of the root and bark of Bahuinia tomentosa is used by the medical practitioners. In India and Srilanka, the root bark is used internally for gastric problems, while the flower is used as a remedy for dysentery and diarrohea [9, 10]. The present study is focused to investigate the pharmacognostic and phytochemical analysis of leaves of Bauhinia tomentosa . EXPERIMENTAL SECTION 2.1-Sample Collection and Authentication The leaves of Bauhinia tomentosa Linn were collected from Villivakkam, Chennai and authenticated by Dr.S. Jayaraman, Director of Plant and Anatomy Research Centre, West Tambaram, Chennai (PARC/2014/2294) 2.2- Pharmacognostic Analysis 2.2.1- Preparation of Sample Care was taken to select healthy plants. The required samples of leaf were cut and removed from the plant and fixed in FAA (Formalin-5ml+ Acetic acid-5ml + 70% Ethyl alcohol-90ml).After 24 hrs of fixing, the specimens were dehydrated with graded series of tertiary –Butyl alcohol [11]. Infiltration of the specimens was carried by gradual addition of paraffin wax (melting point 58-60°C) until TBA solution attained super saturation. The specimens were cast into paraffin blocks. 2.2.2- Sectioning The paraffin embedded specimens were sectioned with the help of rotary Microtome to a thickness of 10-12 µm. Dewaxing of the sections was by customary procedure [12]. The sections were stained with Toluidine blue [13].The staining results were remarkably good; and some cytochemical reactions were also obtained. Wherever necessary, sections were also stained with safranin and fast-green and Iodine (for Starch) For studying the stomatal morphology, venation pattern and trichome distribution, paradermal sections (sections taken parallel to the surface of leaf) as well as clearing of leaf with 5% sodium hydroxide or epidermal peeling by partial maceration employing Jeffrey’s maceration fluid were prepared. Glycerine mounted temporary preparations were made for macerated/cleared materials. Powdered materials of different parts were cleared with sodium hydroxide and mounted in glycerine medium after staining. Different cell components were studied and measured. 2.2.3-Photomicrographs Microscopic descriptions of tissues are supplemented with micrographs wherever necessary. Photographs of different magnifications were taken with Nikon lab photo 2 microscopic Unit. For normal observations bright field was used. For the study of crystals, starch grains and lignified cells, polarized light was employed. Since these structures have birefringent property, under polarized light they appear bright against dark background. Magnifications of the figures are indicated by the scale-bars. Descriptive terms of the anatomical features are as given in the standard Anatomy books [14]. 2.3-Phytochemical Analysis 2.3.1- Preparation of plant extract The leaves of Bauhinia tomentosa Linn. Were first washed, shade dried and then ground into coarse powder using a mechanical grinder. 50 g of the powdered leaf material was extracted with petroleum ether, ethyl acetate, ethanol and water successively by hot percolation method [15] 2.3.2-Qualitative Phytochemical Analysis Qualitative tests for alkaloids, flavonoids, carbohydrates, glycosides, saponins, tannins, proteins and anthraquinones were performed according to the standard procedure [16], Alkaloids : Mayer’s Test: To 1ml of extract, add 2ml of Con.HCL then few drops of Mayer s reagent was added. Green colour or white precipitate shows the presence of alkaloids. Flavonoids : To 1ml of extract add 2ml of 1% aluminium solution. Appearance of yellow colour indicates the presence of flavonoids. Steroids : To 1ml of the filtrate add 10ml of Chloroform and 10ml of sulphuric acid slowly by the sides of the test tube. Red colour in upper layer and sulphuric acid layer showed the yellow colour with green fluorescence shows the presence of steroids. 272 R. Balabhaskar and K. Vijayalakshmi J. Chem. Pharm. Res., 2015, 7(4):271-277 ______________________________________________________________________________ Tannins : To 1ml of extract, add 2ml of 0.1% Ferric chloride. Positive test shows brownish green or blue black colouration Saponins : To 1ml of extract, add 2ml of distilled water and shaken vigorously and allowed to stand for 10 min. There is the development of foam on the surface of the mixture. Then shake for 10 minutes, it indicates the presence of saponins. Anthraquinones : To 1ml of the extract, add 10ml of benzene, filter it and add 5ml of 10% (v/v) ammonia to the extracts and shake well. Development of pinkish coloured solution indicates the presence of anthraquinones. Phenols: To 1ml of extract add 5ml of Folin s ciocalteau reagent followed by 4ml of sodium carbonate Appearance of blue colour shows the presence of phenols. Carbohydrates : a) To 1ml of extract add few drops of Molisch s reagent and few drops of concentrated sulphuric acid which gives purple colour.b) To 1ml of extract, add 5ml of Benedict’s reagent and boil for 5 minutes. Bluish green colour indicates the presence of carbohydrates. Amino acids : To 1ml of filtrate, few drops of 0.2% ninhydrin was added and heated for 5 minutes. Formation of blue colour indicates the presence of amino acids. Cardiac glycosides : To 1ml of extract, add 1ml of Ferric chloride reagent and few drops of concentrated sulphuric acid. Greenish blue colour appears within few minutes indicating presence of cardiac glycosides. RESULTS AND DISCUSSION 3.1 Pharmacognostic Analysis 3.1.1 Anatomy of the Lamina The leaf is dorsiventral and heteromorphic in internal structure. The lateral veinlets (Fig 1.1), lateral vein (Fig 1.2) and the main midrib (Fig 1.3) are prominently projecting on the abaxial side of the lamina. The adaxial side of the veins is flat. The vascular system of the veins includes a single wide and thick vascular strand, which is thick in the mid part and thin on either side (Fig 1.2 and 1.3). The vascular strand consists of several vertical
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