In vivo pharmacokinetics of salvianolic acid B and its metabolite in rats / 中草药

ABSTRACT

Objective: A rapid and sensitive method using liquid chromatography-tandem mass spectroscopy (LC-MS/MS) was developed and validated for the simultaneous quantitative determination of salvianolic acid B (Sal B) and its main active metabolite danshensu(DSS) in rat plasma. Methods: The analytes were extracted by liquid-liquid extraction with ethyl acetate after IS (IS, chloramphenicol) spiked. The separation was performed on a Symmetry C18 column with methanol-acetonitrile-0.5% formic acid (55540) as mobile phase at a flowrate of 0.4 mL/min. The triple quadrupole LC-MS system was operated under selected reaction monitoring (SRM) mode using the electrospray ionization technique in negative mode. Quantification was performed using SRM of the transitions m/z 717→519 for Sal B, 197→135 for DSS, and 321→152 for the IS, respectively. Results: The nominal retention times for Sal B, DSS , and IS were 3.12, 2.60, and 3.98 min, respectively. The standard calibration curve for spiked rat plasma containing Sal B was linear over the range 10-5 000 ng/mL with a correlation coefficient (r >0.995). And DSS was linear over the range 5-5000 ng/mL with a correlation coefficient (r>0.995). The lower limits of quantification (LLOQ) of Sal B and DSS of the method were 10 and 5 ng/mL, respectively. The intra- and inter-day accuracy and precision of the assay were less than 12.6%. After Sal B was ig administered to rats, absorption of Sal B was rapidly metabolized to DSS. Pharmacokinetic parameters of Sal B and DSS after ig administration Sal B to Wistar rats were Cmax (1.21±0.31) and (0.27±0.05) μg/mL, tmax (0.50±0.00) and (0.56±0.18) h, t1/2 (1.20±0.11) and (1.57±0.16) h, AUC0-1 (1.31±0.30) and (0.39±0.05) μg·mL-1·h. Conclusion: This method has been applied successfully to a pharmacokinetic study of Sal B and its metabolite DSS involving the ig administration of Sal B to rats.