Determination of the phytochemical composition of Jingning fang and the in vivo pharmacokinetics of its metabolites in rat plasma by UPLC-MS/MS

- The composition and metabolites of JNF in rat plasma were identified using thermo Q Exactive™ Plus Orbitrap™ mass spectrometer.
- We have developed a sensitive and validated method for measurement of JNF in plasma LC-MS/MS.
- Developed method was successfully applied to pharmacokinetic study in rats.
- Precision and accuracy values in all tests were within the acceptable ranges.

Jingning fang (JNF) is an effective Traditional Chinese Medicine (TCM) which is used for the treatment of Attention Deficit Hyperactivity Disorder (ADHD). To clarify the bioactive constituents of JNF, a Thermo Q Exactive™ Plus Orbitrap™ mass spectrometer was used in this study. More than 127 chemical compounds were isolated and identified tentatively in the JNF extract, while 42 prototype constituents with 4 potential metabolites were identified tentatively in rat plasma. A method for simultaneous determination of polygalaxanthone III (PAIII), sibiricose A5 (A5), sibiricose A6 (A6), 3, 6′-disinapoyl sucrose (3,6′-DISS), tenuifoliside C (TEC), tenuifolin B (TNB), verbascoside (VCE), heterophyllin B (HEB) and schisandrin (SCH) in rat was developed and validated using polydatin (PLN) and psoralen (PSN) as internal standards. All calibration curves proved favorable linearity (R2 ≥ 0.9923) in linear ranges. The lower limit of quantification (LLOQ) was 2.5 ng/mL for PAIII, A5, 3, 6′-DISS, TNB, VCE, HEB and SCH, 1.0 ng/mL for A6 and TEC, respectively. Intra-day and inter-day precisions didn't exceed 14.0% for all the analytes. Extraction recoveries and matrix effects of analytes and IS were acceptable. The validated method has been successfully applied to the pharmacokinetics (PK) studies of the nine compounds in JNF. These findings are useful for predicting the bioactive components of JNF, and will aid in optimizing dose regimens of the drug.

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