Alpha-Amylase Inhibitory Assay
This assay was carried out using a modified procedure of McCue and Shetty [17]. A total of 250 μL of extract (1.25–10 mg/mL) was placed in a tube and 250 μL of 0.02 M sodium phosphate buffer (pH 6.9) containing α-amylase solution (0.5 mg/mL) was added. This solution was preincubated at 25°C for 10 min, after which 250 μL of 1% starch solution in 0.02 M sodium phosphate buffer (pH 6.9) was added at timed intervals and then further incubated at 25°C for 10 min. The reaction was terminated by adding 500 μL of dinitrosalicylic acid (DNS) reagent. The tubes were then incubated in boiling water for 5 min and cooled to room temperature. The reaction mixture was diluted with 5 mL distilled water?? and the absorbance was measured at 540 nm using spectrophotometer.
( cre: Modes of Inhibition of α-Amylase and α-Glucosidase by Aqueous Extract of Morinda lucida Benth Leaf - PMC (nih.gov) )
The absorbance of the solution without dilution would probably be too high to measure by the spectrophotometer. Diluting it brings it into the readable range.
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