Millon's Test Lab Report

Phytochemical tests were carried out on the methanol & ethyl acetate extracts of C.mukulusing standard procedure to identify and to confirm the constituents present in these extracts. (Sofowora 1993).
Test for Alkaloids
Mayer’s test was used to determined alkaloids, 50µl of both methanol and ethyl acetate extracts of C.mukul were treated with 1.36g of mercuric chloride and 5g of potassium iodide in 100ml distilled water/Mayer’s reagent and studied for the formation of cream colour precipitate. Wagner’s reagent: 10-50µl of extract was treated with 1.27g of iodine and 2g of potassium iodide in 100ml distilled water.
Test for flavonoids
NaOH and HCL test was used to detect flavonoids Small amount ofguggulu extract was treated with aqueous NaOH and HCL and observed for the formation of yellow orange colour. H2SO4 test: Extract was treated with conc. H2SO4 and observed for the formation of orange colour for the presence of flavonoids.
Test for proteins
Millon’s test was detect proteins 2ml of Millon’s reagent was mixed with plant extract and observed for the formation of white color which turns red on gentle boiling that confirms the presence of protein.
Test for tannins
Extract was treated with 10% lead acetate and observed for the formation of white color precipitate.

The well of 6 mm diameter punched off with the help of sterile cork borer in the nutrient agar plate and then the volume 100mg/ml of extract was carefully added. Plates were incubated at 4°C for pre-diffusion of extracts in a refrigerator. The plate's were then incubated at 37°C in an incubator for 24 h. The antibacterial activity was evaluated by measuring the diameter of the zone of inhibition excluding the diameter of the well. Amoxicillin at a concentration of 100mg/ml as the positive control and 100% DMSO were used as the negative