Qualitative and Quantitative analysis of Complex Herbal Components in Rat Plasma and Intestine using High Resolution Accurate Mass Spectrometry

Ingredients of traditional Chinese medicine (TCM) play a very important role in the treatment of various diseases such as diabetes, cardiac arrhythmia, cancer, etc. Evaluation of PK profile and biotransformation of active ingredients in TCM will clarify pharmacokinetic and pharmacodynamics effects. Here we describe the advantages of easy and generic methods using a high resolution accurate mass spectrometry (HRMS) technique coupled with ultra high performance chromatography (UPLC) to quantify and evaluate ginsenosides metabolite profile in biological samples. Adult male Zucker rats were dosed daily with Ontario-grown North American ginseng by oral gavage for 42 days to evaluate the effect on vascular and reproductive health. For the present study, plasma samples were protein precipitated and intestinal contents were homogenized to extract ginsenosides and its metabolites. HRMS analysis was performed on TripleTOF™ 5600 system and metabolite ID and structural elucidation was done by MetabolitePilot™ and PeakView™ software. UPLC to HRMS was developed to achieve faster gradient method with 10min run time to separate 6 major active ingredients of ginsenosides (Rg1, Re, Rb1, Rc, Rb2 and Rd) and their metabolites in complex biological matrix. TOFMS for quantitative analysis and information dependent TOF MS/MS for qualitative analysis with fast cycle times was used. Most of the ginsenosides experienced deglycosylation. Rb1 and its deglycosylated metabolites were detected in plasma at high dose (500mg/kg). Concentrations of ginsenosides and its metabolites were high in intestinal samples compared to plasma, thus indicating less bioavailability.