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目的:建立测定人血浆中拉西地平浓度的方法。方法:200μl血浆样本采用500μl叔丁基甲醚萃取浓缩,采用HPLC-MS/MS测定,以~(13)C_8-拉西地平为内标,色谱柱为Agilent ZORBAX Eclipse XDB C_(18)(150 mm×2.1 mm,5μm),流动相为水(含5 mmol·L~(-1)甲酸铵)-乙腈(15∶85,v/v),流速0.3 ml·min~(-1),柱温40℃,进样量10μl,采用电喷雾离子源,以多反应监测方式进行正离子扫描,以[M+NH_4]~+作为分子离子峰,用于拉西地平定量的三对反应离子对为m/z 473.5→m/z 410.3、m/z473.5→m/z 400.1和m/z 473.5→m/z 354.3,用于内标定量的反应离子对为m/z 481.4→m/z 362.3。结果:拉西地平在0.1~(-1)0 ng·ml~(-1)范围内线性关系良好(r>0.996 9),定量下限为0.1 ng·ml~(-1);日内、日间精密度为3.15%~7.04%,相对偏差(RE)为-8.58~12.17%,提取回收率为107%~118%,RSD<15%。血浆样品在经历3次冻融(-20℃到室温)循环和冷冻放置40 d等条件下均稳定,处理后样品在室温和自动进样器中(4℃)各放置24 h均稳定(RSD<15%)。结论:该方法快速、准确、灵敏度高、专属性强,可应用于拉西地平相关的生物等效性一致性评价和药动学研究。
Objective: To establish a method for the determination of lacidipine concentration in human plasma. METHODS: 200μl plasma samples were extracted with 500μl tert-butyl methyl ether and concentrated by HPLC-MS / MS. The internal standard was (13) C_8-raxirizine. The column was Agilent ZORBAX Eclipse XDB C 18 (150 mm × 2.1 mm, 5 μm). The mobile phase consisted of water (containing 5 mmol·L -1 ammonium formate) -acetonitrile (15:85, v / v) at a flow rate of 0.3 ml · min -1 ℃, the injection volume of 10μl, the use of electrospray ionization source, multi-reaction monitoring positive ion scan to [M + NH_4] ~ + as molecular ion peak for the determination of lacidipine quantitative three pairs of reaction ion m / z 473.5 → m / z 410.3, m / z 473.5 → m / z 400.1 and m / z 473.5 → m / z 354.3. The reaction ion pair used for internal standard quantification was m / z 481.4 → m / z 362.3. Results: Laxidipine showed a good linearity (r> 0.996 9) in the range of 0.1-1 ng · ml -1 with a lower limit of quantification of 0.1 ng · ml -1. The precision was from 3.15% to 7.04%, the relative deviation (RE) was from -8.58% to 12.17%, the recovery was from 107% to 118% and the RSD was less than 15%. Plasma samples were stable under three cycles of freezing and thawing (-20 ° C to room temperature) and 40 ° C frozen storage. Samples were stable for 24 h at room temperature and in an autosampler (4 ° C) (RSD <15%). Conclusion: The method is rapid, accurate, sensitive and specific. It can be applied to evaluate the bioequivalence consistency and pharmacokinetics of lacidipine.