Research Center for Drug Metabolism, College of Life Science, Jilin University, Changchun 130021, China.
J Pharm Biomed Anal. 2010 May 1;52(1):149-54. doi: 10.1016/j.jpba.2009.12.012. Epub 2009 Dec 21.
A rapid and sensitive method for the simultaneous quantitation of hydrochlorothiazide (HCT) and metoprolol (MET) in human plasma based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed and validated. MS/MS detection involved switching the electrospray ionization (ESI) mode during chromatography from negative to detect HCT and its internal standard (I.S.) 5-bromouracil to positive to detect MET and its I.S. tramadol. Sample preparation by liquid-liquid extraction with diethyl ether-dichloromethane (60:40, v/v) was followed by chromatography on a Venusil MP-C18 column using methanol-ammonium acetate (10mM)-formic acid (pH 3.4) (50:50:0.05, v/v/v) at a flow rate of 0.8mL/min. The method was linear in the concentration range 3-1000ng/mL for both HCT and MET using 100microL human plasma. Intra- and inter-day precisions (as relative standard deviation) for HCT were 2.9-3.9% and 3.9-4.7%, respectively and for MET were 2.4-4.1% and 4.7-6.2%, respectively. Accuracies (as relative error) were +/-3.8% and +/-2.6% for HCT and MET, respectively. The assay was successfully applied to a pharmacokinetic study involving a single oral dose of a combination tablet (25mg HCT, 50mg MET) in healthy volunteers.
已开发并验证了一种基于液相色谱-串联质谱（LC-MS/MS）的快速灵敏的方法，用于同时定量人血浆中的氢氯噻嗪（HCT）和美托洛尔（MET）。MS/MS 检测涉及在色谱过程中切换电喷雾电离（ESI）模式，从负模式切换为检测 HCT 及其内标（IS）5-溴尿嘧啶，到正模式检测 MET 及其 IS 曲马多。采用二乙醚-二氯甲烷（60:40，v/v）进行液液萃取，然后在 Venusil MP-C18 柱上进行色谱分离，以甲醇-乙酸铵（10mM）-甲酸（pH 3.4）（50:50:0.05，v/v/v）为流动相，流速为 0.8mL/min。该方法在 HCT 和 MET 的浓度范围为 3-1000ng/mL 时，使用 100μL 人血浆具有线性。HCT 的日内和日间精密度（相对标准偏差）分别为 2.9-3.9%和 3.9-4.7%，MET 的分别为 2.4-4.1%和 4.7-6.2%。HCT 和 MET 的准确度（相对误差）分别为 +/-3.8%和 +/-2.6%。该测定方法成功应用于一项健康志愿者单次口服复方片剂（25mg HCT，50mg MET）的药代动力学研究。
