Pharmacokinetic assessment of twenty compounds from Bufei Huoxue capsule in rats using HPLC-MS/MS

Bufei Huoxue capsule (BHC), consisting of Astragali Radix, Paeoniae Radix Rubra, and Psoraleae Fructus, demonstrates remarkable efficacy in treating various pulmonary conditions. In this research, an original approach using high-performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS) was devised to quantitatively determine twenty components (calycosin-7-O-β-D-glucoside, methylnissolin, calycosin, genistein, formononetin, daidzein, angelicin, psoralen, astragaloside IV, 6'-O-galloyl paeoniflorin, benzoylpaeoniflorin, albiflorin, paeoniflorin, oxypaeoniflorin, psoralenoside, isopsoralenoside, catechin, kaempferol, p-coumaric acid, and protocatechuic acid) of BHC in rat plasma. For the twenty selected quantitative prototypes, all calibration curves exhibited excellent linearity (r ≥ 0.9926) within their respective linear ranges. All analytes showed intra-day and inter-day precision (RSD) below 13.41 %, with the accuracy varying between -13.56 % and 12.97 %. The extraction recoveries were measured in the interval of 62.17-96.05 %, and the matrix effects exhibited a range from 74.42 % to 115.96 %. The stability tests indicated that the twenty compounds maintained stability across four distinct storage conditions, and their relative standard deviations did not exceed 14.34 %. Following validation, this methodology was applied to determine the pharmacokinetic behavior of twenty compounds in BHC after oral administration to rats, with relevant pharmacokinetic parameters derived. The obtained data provided a valuable reference for future protocol enhancements and clinical applications.