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

AbstractPolygonum 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.Keywords: Affinity-ultrafiltration (AUF) and high-performance – tandem mass spectrometry (HPLC-MS/MS)molecular dockingPolygonum amplexicaule radixthrombin Disclosure statementNo potential conflict of interest was reported by the author(s).Date availability statementThe data that support the findings of this study are available from the corresponding author upon reasonable request.Additional informationFundingThis study was funded by Natural Science Foundation of Hunan Province (2021JJ30511) and Hunan Administration of Traditional Chinese Medicine (2021062).