Preliminary qualitative phytochemical analysis

Ten (2ml) of alcoholic extract and fractions were stirred with 5 ml of 1% HCL on a steam bath.

Mayer’s (1.35gm mercuric chloride in 60ml water + 5gm potassium iodide in 10ml water )and Wagner’s reagents (1.27g of iodine and 2g of potassium iodide in 100ml of water) were added, white and reddish brown color precipitate respectively, were taken as evidence for the presence of alkaloids

Two tests used for detection of flavonoids

Lead acetate test: to 2ml of alcoholic extract and fractions, 1ml of 10% lead acetate solution was added. The formation of a yellowish- white precipitate was taken as a positive test for flavonoids

NaOH test : 2ml of the extract and fractions were treated with aqueous NaOH and HCl, and looking for the formation of a yellow orange color

Two tests used for identification of steroids

Liebermann-Burchard test : 2ml of the extract and fractions were treated with chloroform, acetic anhydride and drops of sulphuric acid was added. The formation of dark pink or red color indicates the presence of steroids

H2SO4 test ₂SO₄ test: The development of a greenish color was considered as indication for the presence of steroids, when 2 ml of the extract and fractions were treated with sulphuric and acetic acids.

Two ml of the organic extract and fractions were dissolved in 2 ml of chloroform

Evaporated to dryness

2 ml of concentrated sulphuric acid was then added and heated for about 2 min. A grayish color was considered an indication for the presence of terpenoids.