A High-Throughput Assay for Quantification of Starch Hydrolase Inhibition Based on Turbidity Measurement

作者：Liu Tingting; Song Lixia; Wang Hongyu; Huang Dejian^*

来源：Journal of Agricultural and Food Chemistry, 2011, 59(18): 9756-9762.

DOI：10.1021/jf202939d

• alpha-Amylase
• alpha-glucosidase
• amyloglucosidase
• starch hydrolysis
• proanthocyanidins
• area under the curve (AUC)
• turbidity

A high-throughput method for rapid determination of starch hydrolase inhibition was developed using a 96-well microplate UV-vis reader to monitor the turbidity decrease over time. The area under the curve of turbidity measured over time was used to quantify the inhibitory effect of polyphenolic compounds on porcine pancreatic amylase, rat intestine alpha-glucosidase, and fungal amyloglucosidase. Acarbose equivalence (AE) was introduced for the first time and defined as IC(50) of acarbose divided by the IC(50) of the sample measured under the same 96-well plate. This way, the run-to-run variations are canceled out. Among the plant extracts tested, grape seed extracts (1,440 mu molAE/g) and cinnamon bark extracts (1600 mu molAE/g) are the most active in inhibiting rat intestine alpha-glucosidase. For porcine a-amylase inhibition, grape seed extracts (5710 mu mol AE/g) are close to four times more active (equal weight basis) than acarbose (1550 mu molAE/g).

• 出版日期2011-9-28