RESUMO
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 (55:5:40) 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.
RESUMO
OBJECTIVE:To establish a high performenc capillary zone electrophoresis method for the determination of the water-soluble active ingredients in different parts of the Salvia miltiorrhiza.METHODS:Capillary electrophoresis was conducted using uncoated capillary column(75 ?m?50.2 cm,effective length=40 cm)with 5 mmol?L-1 phosphate-borax(ph7.4)as running buffer by pressure injection(0.5 psi/4 s)and constant voltage separation(20 kV)with a detection length of 210 nm and a column temperature of 25 ℃.RESULTS:A complete baseline separation of PAH,DSS and PA was achieved within 8 min under the optimized conditions.The good linear range was from 2.5 ?g?mL-1 to 200.0 ?g?mL-1 for all the three ingredients.The average recoveries of the 3 ingredients were 100.04%,99.99%,and 100.01%,respectively,with RSD at 1.75%,1.73%,and 1.74%,respectively.CONCLUSION:The method is satisfactory in precision,recovery and linearity,and it can be used to determine three water-soluble active ingredients in different parts of the Salvia miltiorrhiza.