高效液相色谱-质谱联用法测定风湿免疫病患者口服低浓度甲氨蝶呤的血药浓度  

Determinatinon the concentration of low-dose methotrexate in patients plasma with rheumatic disease by HPLC-MS/MS

作　　者：吴凡[1,2] 张楠[1,2] 康子胜[3] 许俊羽[1] 赵侠[1] 周颖[1] 崔一民[1] 

机构地区：[1]北京大学第一医院药剂科,北京100034 [2]北京大学药学院药事管理与临床药学系,北京100191 [3]北京大学第一医院临床药理研究所,北京100034

出　　处：《中国临床药理学杂志》2016年第21期1993-1996,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81373487)

摘　　要：目的建立风湿免疫病患者服用甲氨蝶呤的液相色谱-质谱联用血药浓度测定方法。方法采用甲醇沉淀蛋白,以氨基蝶呤作为内标,用Epic C18色谱柱(4.6 mm×50 mm,3μm),甲醇(含0.1%甲酸)-水(含0.1%甲酸)为流动相,梯度洗脱,流速0.6 m L·min^(-1),柱温40℃。用电喷雾离子化源,正离子模式,多反应监测(MRM)扫描方式进行监测。考察方法的专属性、标准曲线、定量下限、精密度、准确度、回收率、基质效应和稳定性。结果甲氨蝶呤在0.5~200.0 ng·m L^(-1)范围内线性良好(r=0.999 2),定量下限为0.5ng·m L^(-1),日内、日间精密度及准确度RSD<15%,回收率为76.80%~85.49%。结论该检测方法快速、灵敏,满足对服用甲氨蝶呤72 h内的风湿免疫病患者血药浓度的准确测定。Objective To determine the methotrexate concentration of patients with rheumatic disease in plasma by HPLC - MS/MS. Methods Plasma samples were prepared by protein precipitation with methanol, while using aminopterin as an internal standard. The chromatographic separation was accomplished on an Epic C18 column （4.6 mm ×50 mm, 3 μm） with a mobile phase composed with methanol and water all containing 0. 1% formic acid under a gradient elution mode. The detection was performed with positive -ion electrospray ionization （ESI） by multiple reactions monitoring （MRM）. The specifici- ty, lower limit of quantitation（LLOQ）, standard curve, precision, accura- cy, recovery, matrix effect and stability were investigated. Results The linearity of this method was found to be within the concentration range of 0. 5 -200. 0 ng mL-1 for methotrexate in plasma （r = 0. 999 2）. The LLOQ was 0. 5 ng mL-l. The relative standard deviations （RSD） of intra and inter precisions were less than 15% and the accuracies were within 15% for methotrexate. The recovery rate was 76. 80% -85.49%. Conclusion This method was rapid and sensitive. It was suitable and accurate for the determination of plasma concentration in rheumatoid disease patients following oral administration methotrexate within 72 hours.

关 键 词：甲氨蝶呤 液相色谱质谱联用 血药浓度 风湿免疫病 

分 类 号：R979.1[医药卫生—药品]