Determination of Anti-Thrombin Active Components in Polygonum Amplexicaule Radix by Affinity-Ultrafiltration (AUF) and High-Performance – Tandem Mass Spectrometry (HPLC-MS/MS)

Abstract

Polygonum Amplexicaule Radix is a traditional Chinese medicine for promoting blood circulation and removing blood stasis, which is often used to treat thrombotic diseases. However, its potential anticoagulant active ingredients have been unexplored. In this study, a method for rapid screening of anticoagulant active ingredients from P. amplexicaule by affinity-ultrafiltration (AUF) and high-performance – tandem mass spectrometry (HPLC-MS/MS) was established, which using thrombin as an affinity receptor target. 5 potential anti-thrombin active components were screened: gallic acid, procyanidin B2-3''O-gallate, 11-O-galloylbergenin, (-)-epicatechin gallate and di-galloyl-O-bergenin by AUF and HPLC-MS/MS. The inhibitory ability of these active ingredients to thrombin was confirmed by in vitro activity experiments and molecular docking, hence justifying that P. amplexicaule is a traditional Chinese medicine with excellent anticoagulation effects. Meanwhile, it has been shown that AUF-MS could be a way rapid screening for active substances.