INTRODUCTION:

Medicinal plants are an important therapeutic aid for various diseases [1]. Medicinal properties of several herbal plants have been documented in ancient Indian literature for the treatment of various disorders. According to W.H.O. report, increasing demand of herbal medicines expected grown to 5 trillion by the year 2050 in international market [2]. Medicinal plants produce secondary metabolites, these components/ chemicals are produced by plants to protect themselves but recent researches demonstrate that they can also protect humans against diseases. These secondary metabolites or active components have curative power against many disorders. Nowadays herbal medicines used by 60% of the world population and 80% population of Africa depend on traditional medicines for primary health care [3]. Here in the present study six plants were taken for the phytochemical screening. The phytochemical constituents were studied by qualitative analysis for performing various chemical tests.

MATERIAL AND METHODS:

Plant material

Plant sample (leaves) were collected from Forest Research Institute, Dehradun, Uttarakhand, India and identified by Dr. S. P. Chaukiyal (Scientist- D) Botany Division FRI Dehradun.

Extraction Methods

The plant samples were washed in tap water, dried, and placed into a blander to be grounded into powder. Three solvents (Methanol, ethanol and water) were used for the soxhlet extraction procedure in different ratios. After 6 to 8 hours, extract is collected, filtered with muslin cloth and transferred to 50 ml tubes and centrifuged for 15 min at 4,000 rpm at 25ºC. the supernatant was collected and kept to dry. After drying, it was mixed with 10% DMSO and used for the experiments.

Phytochemical analysis

Chemical tests for the screening and identification of active components in the leaves extract using standard protocols are as described in [4,5]. For each test, 100 µl of each solvent extract was used for analysis.

Test for Saponins

Extract was taken in a test tube and shaken vigorously. The formation of stable foam was taken as an indication for the presence of saponines.

Test for Phenols

Extract mixed with 2 ml of 2% solution of FeCl₃. Blue/green color indicated the presence of phenols.

Test for Tannins

Extract mixed with 2 ml of 2% solution of FeCl₃. Black color indicated the presence of tannins.

Test for Terpenoids

Extract was mixed with 2 ml of chloroform. Then 2 ml of concentrated Sulfuric acid was added carefully and shaken gently. Reddish brown colors observed in the interphase indicate the presence of terpinoids.

Test for Flavonoids

Extract was treated with few drops of sodium hydroxide solution, and then formation of intense yellow color, which becomes colorless on addition of dilute acid indicates the presence of flavonoids.

Test for Glycosides

Extract was mixed with 2 ml of glacial acetic acid containing few drops of 2% FeCl₃, mixture was poured into another tube containing 2 ml of concentrated sulfuric acids. A brown ring at the inter phase indicates the presence of glycosides.

Test for protein

The extract was treated with few drops of concentrated nitric acid, formation of yellow color indicates the presence of proteins.

Test for Alkaloids

Extract was dissolved individually in diluted HCL and filtered, filters were treated with saturated picric acids and formation of brown precipitate indicates the presence of alkaloids.

Test for Steroids

Extract mixed with 2 ml of chloroform then carefully add H₂SO₄, formation of reddish brown color indicate presence of steroids.

RESULTS AND DISCUSSION:

The study of chemical constituents of the medicinal plants has acquired a lot of importance all over the world. In the present study plant sample collected from FRI Dehradun and were authentified. Then they were dried and powered and subjected to phytochemical screening. Powders were subjected to extraction with ethanol, methanol and distilled water. The qualitative tests for three different solvents were performed. The investigation shown that positive (+) and negative (-) indicates the presence or absence of active components in leaves extract with different solvents like methanol, ethanol and distilled water respectively. These medicinal plant species contain many active components, these secondary metabolites/components used in various disorders for treatment/modulation with minimum side effects. The results were given in table 1 and 2 respectively.

Table 1: Comparative phytochemical study of Aegle marmelos, Saraca asoka, and Syzygium cumini with three different solvents like methanol, ethanol and distilled water.

Components	*Aegle* *marmelos*			*Saraca* *asoka*			*Syzygium* *cumini*
Components	M	E	W	M	E	W	M	E	W
Alkaloids	+	+	-	+	-	-	-	-	+
Flavonoids	+	+	+	+	+	+	+	+	+
Terpenoids	+	-	-	+	-	-	+	-	+
Saponins	+	+	+	+	-	+	+	+	+
Taninns	-	+	-	+	+	+	+	+	+
Phenol	+	+	+	+	+	-	-	+	+
Proteins	+	+	+	-	+	+	-	-	+
Steroids	_	+	-	+	-	-	+	-	-

M: Methanol, E: Ethanol, W: dist. Water and positive (+) shows presence and negative (-) shows absence of the mentioned compounds

Table 2: Comparative phytochemical study of Ginkgo biloba, Cemellia sinensis/Green tea, and Withania somnifera with three different solvents like methanol, ethanol and distilled water.

Components	*Ginkgo biloba*			Cemellia sinensis			*Withania* *somnifera*
Components	M	E	W	M	E	W	M	E	W
Alkaloids	+	+	-	+	-	+	+	+	-
Flavonoids	+	+	+	+	+	+	+	-	+
Terpenoids	+	-	+	-	+	+	+	+	+
Saponins	+	+	+	+	-	+	+	+	+
Taninns	+	+	-	+	-	-	-	-	-
Phenol	+	+	+	+	-	-	-	+	-
Proteins	+	+	-	+	-	-	-	-	+
Steroids	_	+	+	+	+	-	-	+	-

M: Methanol, E: Ethanol, W: dist. Water and positive (+) shows presence and negative (-) shows absence of the mentioned compounds