Preparation of A. heterophyllus seed powder extract (ASPE)
A. heterophyllus seeds were washed to remove adhering impurities.
Later seeds were air dried under sunlight for a week to remove
the moisture completely. The seeds were cut into small pieces and
powdered in a mechanical grinder. The final sieved powder was
used for all the further studies. The different concentrations (2, 4, 6,
8, and 10%, w/v) of seed extracts solution were prepared by boiling
a mixture of different amounts of seed powder in 100 mL of sterile
distilled water in a 500-mL Erlenmeyer flask for 1 h. The solution
was centrifuged at 10,000 rpm for 15 min to remove the unwanted
seed powder residue. The extract volume was adjusted to 100 ml
by adding distilled water and filtered through Whatman no. 1 filter
paper. This ASPE extract was stored at 4 ◦C and was used for further
studies within 2 days.