A Robust UPLC–MS/MS Method for Simultaneous Quantification of 16 Multiclass Constituents in Jin-Deng-Shan-Gen Decoction and Its Application in a Disease-Model Pharmacokinetic Study

Abstract

This study aimed to develop a robust UPLC-MS/MS method to address analytical challenges in quantifying multiclass constituents of Jin-Deng-Shan-Gen Decoction (JDSGD), and to apply it to investigate how acute pharyngitis alters its pharmacokinetic profile. A UPLC-MS/MS method for 16 structurally diverse JDSGD constituents was developed, addressing several technical challenges, comprehensively validated, and applied to a comparative pharmacokinetic study in normal and acute pharyngitis model rats after oral administration. The validated method achieved baseline separation of all analytes within 12 min, with LLOQs ranging from 0.0781 to 11.7 ng/mL. In this study, acute pharyngitis significantly altered the plasma exposure of most JDSGD analytes, accompanied by decreased intestinal mRNA expression of the tight junction proteins zonula occludens-1 and Occludin. In conclusion, this study established a robust UPLC-MS/MS method for the simultaneous quantification of multiple constituents in JDSGD. Its application effectively uncovered disease-induced pharmacokinetic alterations associated with intestinal barrier alterations, bridging analytical science and pathophysiological pharmacology while providing a reliable tool for pharmacokinetic and quality control studies.