超高效液相色谱串联质谱测定人血浆中雷沙吉兰及其代谢物

目的:建立UPLC-MS/MS法同时测定人血浆中雷沙吉兰(PAI)及其代谢物1-氨基茚满(1-AI)的浓度。方法:血浆样品用乙酸乙酯萃取,离心后取上清液氮气吹干,流动相复溶后进行UPLC-MS/MS测定。色谱柱:ACQUITY UPLC BEH HILIC 色谱柱(2.1 mm×50 mm,1.7 μm);流动相:乙腈-5 mmol·L^-1甲酸铵含2%甲酸(90∶10,v/v);流速:0.4 mL·min^-1;柱温:35 ℃;进样量:5 μL。电喷雾离子化(ESI),正离子模式多重反应选择离子检测(MRM),PAI及内标¹³C₃-PAI、1-AI及相应内标¹³C₃-1-AI的检测离子对分别为m/z 172.0→117.0和175.0→120.0、m/z 134.0→117.0和137.0→120.0。结果:血浆中PAI和1-AI线性范围分别为0.02~10 ng·mL^-1(r=0.9983)和0.1~50 ng·mL^-1(r=0.9993)。日内、日间精密度(RSD)均小于15%,准确度(RE)均小于±15%,血浆基质对PAI和1-AI测定无干扰。结论:本研究建立的同时定量测定人血浆中PAI及1-AI的UPLC-MS/MS方法能够快速测定血浆样品,提供准确可靠的分析结果,可用于大批临床样品的分析。

Objective: To establish a UPLC-MS/MS method for simultaneous determination of rasagiline(PAI) and 1-aminoindan(1-AI) in human plasma.Methods: Plasma sample was first extracted by ethyl acetate,the supernatant liquid after centrifugation was dried by nitrogen and analyzed by the UPLC-MS/MS method under following chromatographic conditions:ACQUITY UPLC BEH HILIC(2.1 mm×50 mm,1.7 μm),the mobile phase compositing with 90% acetonitrile and 10% 5 mmol·L^-1 ammonium formate containing 2% formic acid,with a flow rate of 0.4 mL·min^-1,column temperature at 35 ℃,and injected sample volume of 5 μL.The results of positive ion MRM detection of PAI,1-AI and their internal standards were:m/z 172.0→117.0 and 175.0→120.0, m/z 134.0→117.0 and 137.0→120.0,respectively.Results: A good linearity was observed for both PAI and 1-AI respectively in the range of 0.02-10 ng·mL^-1(r=0.9983) and 0.1-50 ng·mL^-1(r=0.9993),intra-and inter-day precision for PAI and 1-AI were less than 15%,method accuracy(RE%) were between-15% and 15%.The plasma matrix showed no effect on the quantification of PAI and 1-AI.Conclusions:The established UPLC-MS/MS method is proved to be effective for the determination of PAI and 1-AI in human plasma.