Analysis of alcohol dehydrogenase inhibitors from Desmodium styracifolium using centrifugal ultrafiltration coupled with HPLC–MS

Abstract

Alcohol dehydrogenase (ADH) inhibitors play an important role in the treatment of human methanol or ethylene glycol poisoning and the sup­pression of acetaldehyde accumulation in alcoholics. In this study, centri­fugal ultrafiltration coupled with high performance liquid chromatography-mass spectrometry (HPLC–MS) was utilized to screen and identify ADH inhibitors from an ethyl acetate extract of Desmodium styracifolium (Osb.) Merr. The experimental conditions of the centrifugal ultrafiltration were optimized. Under the optimum conditions (ADH concentration: 37.5 μg mL-1, incubation time: 90 min, pH 7.0 and temperature: 15 °C), formononetin and aromadendrin were successfully screened and identified from the ethyl acetate extract of D. styra­cifolium. The screening result was verified by ADH inhibition assays. The IC50 values of formononetin and aromadendrin were 70.8 and 84.7 μg mL-1, which were in accordance with their degrees of binding. Aromadendrin was for the first time reported to have inhibitory activity on ADH. This method provides an effective way to screen active compounds from natural products.