Page 26 - RUT Publisher

Life Science Research and Sustainable Development ISBN: 978-98-84663-33-9

days. After 5 days, the extracts were filtered through no.1 Whatman filter paper and stored in air
tight container for further screening.

Qualitative analysis of phytochemicals
Preliminary phytochemical screening was carried out ( Harborne, 1980 )
1. Test for alkaloids (Mayer’s test)
To 1ml of extract, 1 ml of Mayer’s reagent (Potassium iodide solution) was added.
Formation of whitish yellow or cream coloured precipitate indicates the presence of alkaloids.
2. Test for steroids (Libermann Burchard test)
To 1ml of extract, 2ml of acetic anhydride and 2ml of concentrated sulphuric acid were added.
Formation of violet to blue or green colour indicates the presence of steroids.
3. Test for terpenoids (Salkowski test)
To 1 ml of extract, 2ml of chloroform and few drops of sulphuric acid were added.
Formation of reddish brown ring indicates the presence of terpenoids.
4. Test for flavonoids (Alkaline reagent test)
To 1 ml of extract, few drops of dilute ammonium solution and few drops of concentrated
hydrochloric acid were added.
A yellow colouration indicates the presence of flavonoids.
5. Test for saponins (Froth test)
To 1 ml of extract, 5 ml of distilled water was added and shaked vigorously.
Formation of froth indicates the presence of saponins.
6. Test for phenols (Lead Acetate test)
To 1ml of extract, 1 ml of lead acetate solution was added.
Formation of precipitate indicates the presence of phenols.
7. Test for tannins (Lead acetate test)
To 1ml of extract, 1ml of lead acetate was added.
A formation of white precipitate indicates the presence of tannins.
8. Test for cardiac glycosides (Keller killiani test)
To 1ml of extract, 5ml of distilled water was added and evaporated to dryness. Then to the Sample
2ml of glacial acetic acid containing trace amount of ferric chloride solution was added. Then 1ml
of concentrated sulphuric acid was added along the sides of the tube.
Formation of brown ring underlayed with blue colour indicates presence of cardiac glycosides.
9. Test for amino acids (Ninhydrin test)
To the 1ml of sample, 3 to 4 drops of Ninhydrin solution was added and boiled in water bath for
10 minutes.
Formation of purple or blue colour indicates the presence of amino acids.
10. Test for proteins (Biuret test)
To the 1ml of extract, 1ml of 40% sodium hydroxide solution and 2 drops of 1% copper sulphate
solution were added.
Formation of violet colour indicates the presence of proteins.
11. Test for carbohydrates (Barfoed test)
To the 2ml of extract, 1ml of Barfoed’s reagent was added and boiled in water bath for few
minutes.
Formation of reddish brown precipitate indicates the presence of carbohydrates.
12. Test for reducing sugars (Fehling’s test)
To the 1ml of extract, equal quantities of Fehling solution A and B were added and heated.
Formation of brick red precipitate indicates the presence of reducing sugars.

https://jesjalna.org/Zoology-Publications/index.html 18 Department of Zoology, J. E. S. College, Jalna