超高效液相色谱串联质谱法测定人血浆中阿托伐他汀浓度

摘要

目的：建立测定人血浆中阿托伐他汀浓度的超高效液相色谱串联质谱（UPLC - MS / MS）法。方法色谱柱为 Acquity UPLC BEH C18柱（50 mm ×2.1 mm，1.7μm），柱温40℃，流动相为乙腈-含0.01%甲酸的水溶液（70：30），流速为0.3 mL / min，进样量10μL，电喷雾离子化（ESI +）正离子 MRM 扫描。检测离子为 m / z 559.2[M + H]+→440.5[M + H]+（阿托伐他汀），m / z 315.5[M + H]+→109.3[M + H]+（黄体酮），血浆样品采用乙酸乙酯提取，50℃水浴氮气挥干，用黄体酮作为内标定量分析。结果阿托伐他汀血药浓度在0.01~50 ng / mL 线性关系良好（ r =0.9989）。回收率在95%~106%之间，萃取回收率在72%~79%之间（ RSD ﹤4%，n =5），日内、日间精密度的 RSD 均小于5%（ n =5），最低检测质量浓度为0.002 ng / mL。结论该方法准确、灵敏、简便，适用于阿托伐他汀的血药浓度测定。%Objective To establish the ultra performance liquid chromatography - tandem mass spectrometry(UPLC - MS / MS)method for the determination of atorvastatin concentration in human plasma. Methods The UPLC separation was performed on the Acquity UPLC BEH C18 column(50 mm × 2. 1 mm,1. 7 μm),and the column temperature was 40 ℃ . The mobile phase consisted of acetonitrile aque-ous solution containing 0. 01% formic acid(70 : 30). The flow rate was 0. 3 mL / min. The injection volume was 10 μL. The protonated ions of analytes were detected in positive ionization by multiple reactions monitoring(MRM)mode. The detected ion of atorvastatin was m / z 559. 2 [M + H] +→440. 5[M + H] + ,and the internal standard was m / z 315. 5[M + H] +→109. 3 [M + H] +,human plasma samples were extracted with ethyl acetate,50 ℃ water nitrogen volatile stem,with progesterone as the internal standard for conducting the quan-titative analysis. Results The linear range of atorvastatin were 0. 01 - 50 ng / mL( r = 0. 998 9),the recovery rate was 95% - 106% , The extraction recovery rate was 72% - 79%( RSD ﹤ 4% ,n = 5). the intra - day and the inter - day precision RSD ﹤ 5%( n = 5). The lower limit of quantification(LLOQ)of atorvastatin in plasma was 0. 002 ng / mL. Conclusion This method is accurate,sensitive,simple and suitable for the determination of atowastatin concentration in human plasma.