HPLC-MS/MS法研究熊去氧胆酸在健康人体的药代动力学

Study on pharmacokinetics of ursodexycholic acid in healthy volunteers by HPLC-MS/MS

目的建立HPLC-MS/MS法测定人血浆中熊去氧胆酸(利胆药)浓度并研究其在健康人体的药代动力学。方法20例男性健康受试者空腹口服熊去氧胆酸500 mg,采用Shimadzu VP-ODS色谱柱,流动相为A:乙腈-水(含5 mmol.L-1乙酸铵)(35∶65);B:乙腈;选择(ESI)负离子检测,以盐酸西替利嗪为内标。用高效液相色谱法-串联质谱法测定熊去氧胆酸的血药浓度,并计算药代动力学参数。结果线性范围为40.14～1.20×104ng.mL-1(γ=0.995),最低检测限为40.14 ng.mL-1,低、中、高浓度的日内日间差异均小于20%,相对回收率为90.46%、86.51%和93.41%。口服熊去氧胆酸500 mg后,Cmax为(7.18±2.40)μg.mL-1,tmax为(2.80±0.90)h,AUC0-10为(22.72±6.08)μg.h.mL-1,AUC0-∞为(23.80±7.60)μg.h.mL-1。结论此方法选择性强、灵敏度高、操作简便,适于熊去氧胆酸药代动力学研究。

Objective To establish a HPLC-MS/MS method for determination of ursodexycholic acid in healthy human plasma and study on its pharmacokinetics in healthy human. Methods Twenty male healthy vounlteers were given a single oral dose of ursodexycholic acid 500 mg. Plasma concentrations were determined by HPLC-MS-MS method. The separation was carried out using the Shimadzu VP-ODS column with the mobile phase consisted of A ： acetonitrile-H2O （35：65 ） ; B ： acetonitrile. The cetirizine was taken as the internal standard. And the plasma concentrations were detected in FSI negative selected ion monitoring mode. Results The standard curve was linear in range of 40. 14 - 1.20 × 10^4 ng.mL^-1 （γ =0. 995）. The detection limited in the plasma was 40. 14 ng.mL^-1. RSD of intra-day and inter-day assays were all less than 20% in low concentrations, all less than 15% in middle or high concentrations, while the relativity recovery were 90. 46% , 86.51% and 93.41% respectively. The pharmacokinetic parameters were as follows： Cmax was （7. 18 ± 2. 40 ） μg.mL^-1 , tmax was （ 2. 80 ± 0. 90 ） h, AUC0-10 was （22.72 ±6.08） μg.h.mL^-1,AUC0-∞ was （23.80 ±7.60） μg. h. mL^-1. Conclusion HPLC-MS/MS method is selective, sensitive and convenient, proved to be suitable for determining concentration of ursodexycholic acid in plasma as well as pharmacokinetics studies.