Estimation of alkaloids:

Alkaloids were estimated  following the method of Harborne (1973). Five gm of the plant tissue was weighed and 200 ml of 10% acetic acid in ethanol was added and covered then allowed to stand for 4 hours. This was filtered and the extract was concentrated on a water bath to one-quarter of the original volume. Concentrated ammonium hydroxide solution was added dropwise to the extract until the precipitation was completed. The whole solution was allowed to settle down and the precipitate was collected and washed with diluted ammonium hydroxide solution and then filtered. The residue, that represents the alkaloids, was dried and weighed to be expressed as percentage.

If you are finding following the method difficult, send the plant materials. We shall conduct the analysis and send you the results.  Philippa