2.5. ANTIOXIDANT STATUS
2.5.1. TLC - Semi Quantitative DPPH Assay
0.2 % DPPH solution in methanol was prepared and kept in the fridge for further use. The grid space was marked with 1.0 cm2 space on an aluminum based TLC sheet (Merck silica gel 60F254) and a stock solution of all the extracts together with the standard were prepared in methanol. A series of dilutions of the stock together with the standard were prepared ranging from 400 µL to 0.01µL for the last dilution. The grid on the TLC sheet was labeled with extract on the horizontal axis and amount of extract on the vertical axis. The extracts of different concentrations and the standards were plotted on the TLC sheets and the spots allowed to dry for at least 2 hours. Care was taken to keep the volume of the extracts spotted the same so that all the spots were of the same size for a fair comparison. The sheet was sprayed with 0.2% DPPH solution and the …show more content…
5 mL of 90% aqueous methanol and 0.5 mL Folin-Ciocalteu reagent were added to 0.5 mL of each of the standard solutions and to 0.5 mL of each extract solution (1 mg/mL) in screw cap test tubes. After 3 min, 1 mL of 2% Na2CO3 was then added to each test-tube and the mixture was vigorously shaken for 2 minutes and left to stand for 2 hours at room temperature. The absorbance of the supernatant solution was determined at 725 nm using 90% aqueous methanol as a solvent blank. A gallic acid standard curve was prepared and the equation derived by linear regression (y =36.84 x+0.1069) was used to determine the TPC of each extract in mg of gallic acid equivalents/g of extract (mg GAE/g). The experiment was performed in triplicate and TPC was reported as the average value of 3 trials ± the standard
2.5.1. TLC - Semi Quantitative DPPH Assay
0.2 % DPPH solution in methanol was prepared and kept in the fridge for further use. The grid space was marked with 1.0 cm2 space on an aluminum based TLC sheet (Merck silica gel 60F254) and a stock solution of all the extracts together with the standard were prepared in methanol. A series of dilutions of the stock together with the standard were prepared ranging from 400 µL to 0.01µL for the last dilution. The grid on the TLC sheet was labeled with extract on the horizontal axis and amount of extract on the vertical axis. The extracts of different concentrations and the standards were plotted on the TLC sheets and the spots allowed to dry for at least 2 hours. Care was taken to keep the volume of the extracts spotted the same so that all the spots were of the same size for a fair comparison. The sheet was sprayed with 0.2% DPPH solution and the …show more content…
5 mL of 90% aqueous methanol and 0.5 mL Folin-Ciocalteu reagent were added to 0.5 mL of each of the standard solutions and to 0.5 mL of each extract solution (1 mg/mL) in screw cap test tubes. After 3 min, 1 mL of 2% Na2CO3 was then added to each test-tube and the mixture was vigorously shaken for 2 minutes and left to stand for 2 hours at room temperature. The absorbance of the supernatant solution was determined at 725 nm using 90% aqueous methanol as a solvent blank. A gallic acid standard curve was prepared and the equation derived by linear regression (y =36.84 x+0.1069) was used to determine the TPC of each extract in mg of gallic acid equivalents/g of extract (mg GAE/g). The experiment was performed in triplicate and TPC was reported as the average value of 3 trials ± the standard