The ethanolic soluble fraction (5 g) was fractionated over the silicagel (60-120 mesh) columneluted with chloroform and acetone with increasing acetone concentration 21 fractions werecollected. Test for steroid were performed fractioned 7-10 gives positive test were mixedtogether

Tests for steroid:

Salkowski reaction: A few crystals were dissolved in chloroform and a few drops ofconcentrated sulfuric acid were added to thesolution. A reddish color was seen in the upper chloroform layer .

Liebermann- burchardreaction:

A few crystals were dissolved in chloroform and a few drops of concentrated sulfuric

acid were added to it followed by addition of 23 drops of acetic anhydride.Solution turned violet blue and finally green .

Test for Steroids:

A small portion of extract was dissolved in 1ml of chloroform and filtered. To the filtrateon ice, 1 ml of acetic acid was added and then a few drops of conc. sulphuric acid were rundown the side of the test tube. The appearance of blue, bluish-green or a rapid change from pinkto blue colours indicates the presence of steroids and a combination of pink and these coloursindicates the presence of both steroids & terpenoids.

Test for steroidsLibermann Burchard Reaction:2 ml extract was mixed with chloroform. To this 1-2 ml acetic anhydride and 2 drops

concentrated sulphuric acid were added from the side of test tube. First red, then blue andfinally green colour appears.