Integrating 3D Chromatography With Orbitrap Mass Spectrometry to Profile the Chemical Constituents in JTTZ Formula and Investigate Plasma Pharmacokinetic Changes in Obese Rats

ABSTRACT Introduction Chemical profiling and exposure of multicomponent Chinese medicine formula (CMF) are essential for understanding the multitarget treatment mechanism, as they consist of bioactive substances with varying polarities, molecular weights, and charges. However, the chemical composition of CMF is highly intricate, primarily due to the presence of multiple herbs, rendering chemical identification a challenging task. Objective Jiangtang Tiaozhi formula (JTTZ), commonly used for early diabetes intervention, was chosen to validate our standardized analytical strategy integrating offline three ‐ dimensional (3D) chromatography–DDA (data‐dependent acquisition) with a precursor ion list (PIL)‐comparative pharmacokinetic study. Materials and Methods We have implemented an offline 3D chromatography coupled with data‐dependent MS2 acquisition, including PIL, to obtain comprehensive compound information within JTTZ. We then utilized the MRM quantification approach to determine the pharmacokinetics of the 13 major circulating constituents identified in JTTZ. Finally, the variations in pharmacokinetic parameters between high‐fat diet (HFD) and normal chow diet (NCD) rat models are further illustrated by comparative pharmacokinetics. Result Using an offline 3D chromatography platform, 186 compounds were identified in JTTZ, as opposed to the 68 compounds detected with classical one‐dimensional (1D) LC–MS technology. Through the chemical profiling results of the multidimensional chromatography, we found 41 components of JTTZ absorbed in plasma. We then utilized the MRM quantification approach to determine the pharmacokinetics of the 13 major circulating constituents identified in JTTZ. Finally, the variations in pharmacokinetic parameters between HFD and normal chow diet (NCD) rat models are further illustrated by comparative pharmacokinetics. In the HFD group, the T 1/2 values of epiberberine were higher than those in the NCD group. Conclusion The results show that the pathological state caused by a HFD changed the plasma pharmacokinetics of jatrorrhizine, mangiferin, and timosaponin BII in JTTZ, providing drug exposure evidence for the involvement of JTTZ in early diabetes intervention. The stepwise pipeline for the “offline 3D chromatography‐DDA acquisition method with PIL‐comparative pharmacokinetic study” in the case study of JTTZ has the potential to reveal the exposure profile of other CMFs, from compound identification to pharmacokinetic analysis.