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

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。

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

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 temperature was 40 ℃.Electrospray ionization source(ESI+) was applied.The MS/MS detection was carried 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.