LC-MS/MS同时测定大鼠血浆中红景天苷和酪醇及其药物动力学研究

Simultaneous Determination of Salidroside and Tyrosol in Rat Plasma by LC-MS/MS and Application to Their Pharmacokinetic Study

目的建立大鼠血浆中红景天苷及其苷元酪醇的测定方法,并研究其药物动力学。方法取大鼠10只灌胃给予红景天苷100 mg.kg-1,检测给药前和给药后24 h内红景天苷及酪醇的血浆浓度,并计算其药动学参数。采用液相色谱-串联质谱法,以水杨苷为内标,色谱柱为Alltima C18,流动相:甲醇-水(80∶20),等度洗脱,流速为0.3 mL.min-1,柱温为40℃,电喷雾负离子源,红景天苷、酪醇和水杨苷的选择检测离子质荷比(m/z)分别为299.2→119.6,137.1→119.0和285.1→122.9。结果红景天苷和酪醇检测浓度的线性范围分别为50～5000(r=0.9991)、5～500(r=0.9994)ng.mL-1,最低检测限分别为6.25,2.5 ng.mL-1;药动学参数:红景天苷的t1/2β为(5.67±0.84)h,Cmax为(3914.7±915.8),AUC0→24 h为(8434.2±213.8)ng.h.mL-1;酪醇的t1/2β为(6.24±0.91)h,Cmax为(289.3±44.6)ng.mL-1,AUC0→24 h为(1236.7±73.4)ng.h.mL-1。结论本方法专属性强、灵敏度高、准确性好,可用于红景天苷和酪醇的血药浓度测定,红景天苷和酪醇在大鼠体内的药动学符合二室模型特征。

Objective To establish a method to determine the plasma concentration of salidroside and its aglycone metabolite tyrosol in rat by using LC-MS/MS, and to calculate the pharmaeokinetic parameters of salidroside and its aglycone metabolite tyrosol in rat after oral administration of 100 mg·kg-1 salidroside. Methods Ten rats were given intragastric garage of 100 mg＇kg-1 salidroside, the plasma concentrations of salidroside and tyrosol were detected before administration and within 24 hours after administration, and then the pharmacokinetics parameters were examined. LC-MS/MS was performed on Alhima C18（ 100 ± 2.0 mm, 5μ m） column with salicin as the internal standard. Mobile phase consisted of 80 % methanol-water solution, and flow rate was 0.3 mL min-1. The temperature of column was 40℃. The LC-MS/MS system was operated by using an electrospray ionization probe in the negative ion mode. Scan mode was in multiple reaction ion monitoring （MRM） mode. The ion of monitor was m/z 299.2---119.6 for salidroside, m/z 137.1--119.0 for tyrosol, and m/z 285.1---122.9 for salicin （internal standard） respectively. Results The linear ranges of salidroside and tyrosol were 50-5000 ng·mL-1 （r=0.9991） and 5-500 ng·mL- （r=0.9994） respectively. The lowest limit of quantification （LLOQ） of salidroside and tyrosol were 6.25 ng·mL-01 and 2.5 ng·mL-1. The pharma- cokinetic parameters of t1/2β, Cmax, and AUC0→24h were as follows： （5.67 ± 0.84）h, （3914.7 ± 915.8）ng·mL-1, and （8434.2 ± 213.8）ng·h·mL-1 for salidroside, and（6.24 ± 0.91）h, （289.3 ± 44.6）ng·mL-1, and（ 1236.7 ± 73.4）ng·h·mL-1 for tyrosol. Conclusion A sensitive, accuracy and suitable LC-MS/MS method for determining salidroside and tyrosol has been developed and successfully applied to the pharmacokinetic study of salidroside after oral administration in rats.