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Harika Kotipalli Et Al, J. Global Trends Pharm Sci, 2017; 8(2): 3860 - 3867 Harika Kotipalli et al, J. Global Trends Pharm Sci, 2017; 8(2): 3860 - 3867 An Elsevier Indexed Journal ISSN-2230-7346 Journal of Global Trends in Pharmaceutical Sciences QUALITATIVE PHYSICOCHEMICAL, PHYTOCHEMICAL ANALYSIS AND QUANTITATIVE ESTIMATION OF TOTAL PHENOLS, FLAVONOIDS AND ALKALOIDS OF CASSIA GRANDIS Kotipalli Harika*, Battu Ganga Rao and Devarakonda Ramadevi A.U College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, 530-003, India *Corresponding author E-mail: [email protected] ARTICLE INFO ABSTRACT The aim of present study was to investigate the physicochemical, Key Words phyto-constituents present within the hexane, ethylacetate and methanol extract of Cassia grandis and to estimate the total Cassia grandis, phenolic, flavonoid and alkaloid contents. The amount of total Physicochemical and phenols, were analyzed using a spectrophotometric technique, Phytochemical analysis, based on Folin-ciocalteau reagent. Gallic acid was used as standard Total Phenols, Flavonoids and alkaloids. compound and the total phenols were expressed as mg / g gallic acid equivalents (Standard curve equation: y= 0.0106x + 0.041, R2 = 0.996). Total flavonoid contents of the plant were determined by using quercetin reference standard method. The total alkaloid content was determined by using rutin as standard. Phytochemical analysis indicated the presence of cardiac glycosides, reducing sugars, flavonoids, phenolic compounds and alkaloids. The Physicochemical analysis includes Total Ash (0.072 to 0.666), Water insoluble Ash (0.053 to 0.551), Water soluble Ash (0.019 to 0.170), Acid insoluble Ash (0.033 to 1.946) and Loss on drying (0.471 to 0.924) as per standard methods. INTRODUCTION: Cassia grandis, commonly known as edible, sweet in taste and foul smelling, and coral shower tree, is a semi-deciduous tree, possesses laxative properties.[2] Cassia growing up to a height of about 18 m. The grandis seeds are elliptic, oblong-obovate, tree bears a high, irregular, spreading crown, obovate or obovoid-ellipsoid, with slightly made from dangling branches. Cassia emerginate base. The seeds have biconvex grandis leaves are paripinnate, with 10-20 cross section and are ventrally flattened, leaflets, measuring 3-6 cm in length, and enclosed in a light brown, cartaceous, obtuse or rounded at base and apex. Flower smooth and opaque seed coat. The tree is are in long, drooping, pink or purple generally planted for its ornamental value, coloured axillary racemes, without bracts and the heavy, hard wood is also used in subtending the pedicels. Pods are construction. Fruits ripen in summer. The compresssed-cylindrical, glabrous and sweet pulp and bad smelling of fruit is edible transversely rugose. The pulp of the pods is and used as laxative. 3860 Harika Kotipalli et al, J. Global Trends Pharm Sci, 2017; 8(2): 3860 - 3867 Kingdom Plantae above the flame. Heat till vapours almost cease to be evolved and tare the sample. Sub kingdom Tracheobionta Super Spermatophyta Cool in desiccators. Weigh the ash and division calculate the percentage of total ash with Division Magnoliophyta reference to the air dried sample of crude Class Magnoliopsida drug. Subclass Rosidae Order Fabales Determination of Acid insoluble ash: Family Fabaceae Genus Cassia Using 25 ml of dil. hydrochloric acid, wash Species Cassia grandis the ash from dish for total ash into 1000 ml Taxonomical classification: [3] of beaker. Place a mere gauze over a Bunsen burner. Boil for 5 min. Filter through ash Extraction process less filer paper, wash the residue twice with hot water. Ignite the crucible in the flame and Cool in a desiccator. Weigh and The collected leaves and stem bark were calculate the percentage of acid insoluble dried under shade and powdered. The ash with reference to the air dried sample of powdered materials were carried out crude drug. successive maceration using different solvents such as hexane, ethyl acetate and Determination of water soluble ash: methanol. This is determined in a similar way to acid Procedure: insoluble ash, using 25 ml of water in place Plant material (Crushed or cut small or of dilute hydrochloric acid. Boil for 5 min. moderately coarse powder) was placed in a Filter through ash less filter paper wash the closed vessel. Allowed to stand for seven residue twice with hot water. Cool in days shaking occasionally and Liquid desiccator. Weigh and calculate the strained off. Solid residue (marc) pressed percentage of water soluble ash with (recover as much as occluded solution) reference to the air dried sample of crude Strained and expressed (liquids mixed). drug. Clarified by subsidence or filtration. Then Evaporation and concentration. The extract Loss on drying: thus obtained were concentrated and dried completely, weighed and stored in a desiccators. Procedure: Weight about 5 g of powder into a porcelain Physico-chemical studies: dish. Dry in oven at 100 & 1050c Cool in desiccator. Weigh and the loss in weight Ash values: Used to determine quality and recorded as moisture content. Calculate the purity of crude drug. And to establish the percentage of loss of drying recorded as identity of it. Also used to determine foreign moisture of sample. inorganic matter present as impurity. Swelling index: Determination of total ash: It gives an idea about the mucilage content Weigh about 2 g of powder into a porcelain of the crude drug. Hence it is useful in the dish. Heat with a burner using a flame about evaluation of crude drugs containing 2cm high and supporting the dish about 7cm mucilage. Procedure: 3861 Harika Kotipalli et al, J. Global Trends Pharm Sci, 2017; 8(2): 3860 - 3867 Weigh about 1 g of powder into a 50 ml Filter into a 50 ml of measuring measuring cylinder. Add water up to 40 ml cylinder. Transfer into filtrate in thin making. Shaking occasionally during 24 hrs. porcelain dish. Keep a side for one hour. Measure the The fitrate evaporated to dryness in a volume occupied by swollen powder. tarred flat bottom dish. Weighed and calculate the % w/w of Foaming index: It is evaluated by extractive values of different measuring the foaming ability in term of solvents foaming index. a) If the height of the foam in every Flourescence analysis [4][5] tube is less than 1 cm it means foaming index is less than 100. If the Fluorescence analysis of the drug was height of foam is more than 1 cm observed in UV light (245nm) using various every test tube; the foaming index is extract of the drug. The drug powder was over 1000. In this case, repeat the treated separately with different solutions. experiment using a new series of dilutions o the decoction in order to Phytochemical Analysis get a result. b) If the height of 1 cm in any tube, the The prepared extract was tested for the type volume of the plant material of chemical constituents present by known decoction in this tube(a) is used for qualitative tests. *The following tests were determination of foaming index carried out on the extracts to detect various using formula follows : phytoconstituents present in them. Foaming index = 100/a 1. Test For Alkaloids [6][7] Extractive values: About 50mg of solvent-free extract was Useful in evaluation of crude drug. stirred with little quantity of dilute To determine give idea of nature of hydrochloric acid and filtered. The filtrate chemical constituents present in was tested carefully with various alkaloidal crude drug. reagents as follows. Useful of estimation of constituents extracted with solvent used for a) Mayer’s test: To a few ml of filtrate, extraction. two drops of mayer’s reagent was added along with the sides of test tube. If the Employed for material for which as test is positive, it gives white or creamy yet no suitable chemical or precipitate. biological assay exists. b) Wagner’s test: To a few ml of the Procedure filtrate, few drops of Wagner’s reagent were added along with the sides of the Weigh about 1 g of powder in add test tube. Formation of reddish brown different solvents like (Hexane, precipitate confirms the test as positive. Dichloromethane, chloroform, ethyl c) Hager’s test: To a few ml of filtrate 1 acetate, acetone, methanol, water, or 2 ml of Hager’s reagent was added. A petroleum ether) into a boiling tubes. prominent yellow precipitate indicates Cork the flask and set aside for 24 positive test. hrs. d) Dragendroff’s test: To a few ml of Shaking frequently occasionally filtrate, 1 or 2 ml of Dragendroff’s during 6 hrs and allowed to stand for reagent was added. A prominent reddish 24 hrs. brown precipitate indicates positive test. 2. Test For Carbohydrates [8] 3862 Harika Kotipalli et al, J. Global Trends Pharm Sci, 2017; 8(2): 3860 - 3867 About 100mg of the extract was dissolved in 4. Test For Saponins[11,12] 5ml of distilled water and filtered. The filtrate was subjected to the following tests. a) Froth test: A small quantity of the extract was diluted with distilled water a) Molisch’s test: To 2 ml of filtrate, two to 20 ml. The suspension was shaken in drops of alcoholic solution of α- napthol graduated cylinder for 15 minutes. A was added. The mixture was shaken well two centimeter layer of foam or froth and 1 ml of concentrated sulphuric acid which is stable for 10 minutes indicates was added slowly along the sides of the the presence of saponins. test tube, the test tube was cooled in ice water and allowed to stand. A violet ring 5. Test For Phytosterols And at the junction of two liquids indicates Triterpenoids[13] the presence of carbohydrates b) Fehling’s test: 1 ml of filtrate was a) Liebermann- burchard ‘s test: The boiled on a water bath with 1 ml each of extract was dissolved in acetic anhydride, Fehling’s solution A and B.
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