基于LC-MS/MS法达比加群酯用于房颤患者的治疗药物监测

Therapeutic drug monitoring based on LC-MS/MS method dabigatran etexilate for patients with atrial fibrillation

摘要

目的 建立并验证液质联用技术(LC-MS/MS)测定人乙二胺四乙酸二钾(dipotassiumethylenediaminetetraacetate,K₂EDTA)抗凝血浆中达比加群浓度的分析方法,并将此方法应用于房颤患者服用达比加群酯后药物浓度监测。方法 采用电喷雾正离子化方式,利用多反应监测(multiplereactionmonitoring,MRM)方法检测达比加群及其同位素内标。血浆样品经蛋白沉淀处理后在反相色谱条件下进行分离。色谱柱为Luna C₈柱(50 mm×2.0 mm,3μm),流动相为乙腈(含体积分数0.1%甲酸溶液)和5 mmol·L^-1醋酸铵水溶液(含体积分数0.1%甲酸溶液)进行梯度洗脱。结果 该方法的线性范围为1.00~500 ng·mL^-1(r>0.996),批内和批间精密度(RSD)为1.78%~11.47%,准确度偏差(RE)为-7.29%~1.31%。待测物提取回收率在79.69%~86.31%内,基质效应的变异系数(CV)在1.19%~2.78%内。结论 该方法具有分析速度快、选择性强、准确度高、线性范围宽等优点,成功应用于房颤患者口服达比加群酯后的治疗药物监测。

Abstract

Objective To establish and validate an LC-MS/MS method for the determination of dabigatran in human dipotassiumethylenediaminetetraacetate (K₂EDTA) anticoagulant plasma and to apply this method to the therapeutic drug monitoring of dabigatran etexilate in patients with atrial fibrillation. Methods Dabigatran and its isotope internal standard were ionized in positive ESI source. They were detected by multiple reaction monitoring (MRM) mode. After protein precipitation with acetonitrile, the sample was separated on a Luna C₈ column (50 mm × 2.0 mm, 3μm) with gradient elution. The mobile phase was consisted of acetonitrile (containing 0.1% formic acid) and 5 mmol·L^-1 ammonium acetate aqueous solution (containing 0.1% formic acid). Results The linear range was from 1.00 to 500 ng·mL^-1 with regression coefficient r greater than 0.996. The intra and inter-batch precision ranged from 1.78% to 11.47%. The accuracy deviation was from -7.29% to 1.31%. The extract recovery of dabigatran was in the range of 79.69%-86.31% and the coefficient of variation (CV) of matrix effect was within 1.19%-2.78%. Conclusion This method showed advantages of rapid analysis, strong selectivity, high accuracy and wide linearity. It has been successfully applied to therapeutic drug monitoring of dabigatran etexilate in patients with atrial fibrillation.

关键词

达比加群酯 / 达比加群 / LC-MS/MS / 治疗药物监测 / 房颤患者

Key words

dabigatran etexilate / dabigatran / LC-MS/MS / therapeutic drug monitoring / patients with atrial fibrillation

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