Integrated UHPLC-MS/MS and HRMS Strategy for Screening Potentially Hepatotoxic Furan-Containing Compounds in Dioscorea bulbifera L.

Rong Tan; Ying Liu; Yunfang Jiang; Xin Zha; Jiamin Xin; Cong Wang; Jinghua Ruan; Weiwei Li; Shiyu Zhang

doi:10.3791/70865

A subscription to JoVE is required to view this content. Sign in or start your free trial.

Method Article

Integrated UHPLC-MS/MS and HRMS Strategy for Screening Potentially Hepatotoxic Furan-Containing Compounds in Dioscorea bulbifera L.

171 views

⸱

DOI:

10.3791/70865

⸱

May 19th, 2026

Rong Tan¹^,² , Ying Liu¹^,² , Yunfang Jiang³ , Xin Zha¹ , Jiamin Xin¹ , Cong Wang¹^,² , Jinghua Ruan¹^,² , Weiwei Li⁴ , Shiyu Zhang¹^,²

¹The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, ²School of Pharmacy, Guizhou University of Traditional Chinese Medicine, ³School of Medicine and Health Management, Guizhou Medical University, ⁴Guizhou Provincial Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Guizhou Medical University

Summary

This study developed an integrated strategy combining UHPLC–MS/MS and HRMS to screen for potentially hepatotoxic furan-containing compounds in Dioscorea bulbifera L.

Abstract

Dioscorea bulbifera L. (DBL), a traditional herbal medicine used to treat thyroid disorders and tumors, has raised considerable safety concerns due to its potential hepatotoxic effects. Previous research suggests that this liver injury may be associated with the metabolic activation of furan-containing compounds (FCCs) present in DBL. Here, we systematically characterized FCCs and their reactive metabolites in DBL extract through an integrated analytical approach combining ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and ultra-high performance liquid chromatography coupled with tandem high-resolution mass spectrometry (UHPLC-HRMS). Employing both targeted and untargeted mass spectrometric analyses, we identified a total of 849 constituents in the aqueous extract of DBL, including 10 furanoditerpenoids and 17 additional FCCs. To elucidate the metabolic activation pathways, we used glutathione (GSH) and 4-bromobenzylamine (BBA) as dual trapping agents within a mouse liver microsomal (MLM) incubation system. Through P450-mediated metabolism, seven corresponding adducts derived from diosbulbin B (DSB), C (DSC), D (DSD), E (DSE), L (DSL), N (DSN), and 2-pentylfuran were successfully trapped and characterized. This study establishes a sensitive and specific approach for the comprehensive profiling of potentially hepatotoxic furan compounds in DBL, and provides valuable insights into the formation of reactive metabolites and toxicity mechanisms related to furan-containing herbal medicine.

Introduction

Safety concerns related to traditional Chinese medicine (TCM), especially drug-induced liver injury (DILI), hinder their widespread clinical utility and modernization¹^,²^,³. Growing evidence suggests that most constituents of TCM are not intrinsically toxic; instead, their hepatotoxicity results from metabolic activation in vivo¹^,⁴^,⁵. Furan-containing compounds (FCCs) are well recognized as prominent hepatotoxic precursors, undergoing P450–mediated metabolic activation ....