超高效液相色谱-串联质谱法测定大鼠血浆中替诺福韦的浓度

Determination of Tenofovir Concentration in Rats Plasma by UPLC-MS/MS

目的:建立大鼠血浆中替诺福韦浓度的测定方法。方法:采用超高效液相色谱-串联质谱法。取6只大鼠单次灌胃给予富马酸替诺福韦酯100mg.kg-1,检测给药45min后血浆中药物浓度,以恩替卡韦为内标,选用阳离子固相萃取柱除去血浆中的蛋白质。色谱柱为AcquitybenUPLC-C18,流动相为0.2%甲酸水溶液(含40mmo.lL-1醋酸铵)-乙腈=97:3,流速为0.25mL.min-1,柱温为40℃,电喷雾正离子(ESI+)源,监测离子对分别为质荷比(m/z)287.9→175.7(替诺福韦)和m/z277.9→151.7(恩替卡韦)。结果:替诺福韦检测浓度的线性范围为10～800ng.mL-1(r=0.9992),最低检测限为10ng.mL-1,回收率为(100.22±9.47)%～(102.76±4.99)%,日间、日内RSD均小于9.4%;给药45min后血浆样品中替诺福韦的平均浓度为819.2ng.mL-1。结论:该方法操作简便、专属性强、灵敏度高,可用于血浆中替诺福韦的浓度测定。

OBJECTIVE： To establish the method for the determination of tenofovir concentration in rat plasma. METHODS： UPLC-MS/MS was adopted. 6 rats were given single intragastric administration of tenofovir disoproxil fumarate 100 mg. kg-1. Drug concentration in plasma 45 min after administration was determined. Plasma protein was removed by cation solid phase extraction using entecavir as internal standard. The separation was performed on Acquity ben UPLC-C18 column with a mobile phase consisted of 0.2 % formic acid （containing 40 mmol. L-1 ammonium）-acetonitrile （97：3） at the flow rate of 0.25 mL. min-1. The column tem- perature was 40℃. Electrospray ionization source（ESF） was applied. The MS/MS detection was carded out by monitoring the fragmentation of m/z 287.9→175.7 for tenofovir and m/z 277.9→151.7 for entecavir. RESULTS： The linear range of tenofovir was 10- 800 ng.mL-1（r=0.999 2）, and the lowest detection limit was 10 ng.mL-1. Relative recoveries were （100.22 ± 9.47）%-（102.76 ± 4.99） % ; RSD of intra-day and inter-day were both less than 9.4%. Average concentration of tenofovir in plasma sample 45 min after administration was 819.2 ng.mL-1.CONCLUSION： The method is simple, specific and sensitive, and suitable for the determination of tenofovir concentration in plasma.