液相色谱-质谱联用法测定人血浆中富马酸单甲酯药物浓度及应用  

Determination of monomethyl fumarate in human plasma by HPLC-MS/MS and its application

作　　者：冯湘玲 曹艳 朱瑜 郑恒[1] FENG Xiang-ling;CAO Yan;ZHU Yu;ZHENG Heng(Department of Pharmacy,Tongji Hospital Affiliated with Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院药学部,湖北武汉430030

出　　处：《中国医院药学杂志》2020年第20期2132-2136,2178,共6页Chinese Journal of Hospital Pharmacy

摘　　要：目的:建立液相色谱-质谱联用(HPLC-MS/MS)法测定人血浆中富马酸单甲酯的方法,并应用于检测中国健康受试者血浆中富马酸单甲酯的浓度。方法:血浆样品用醋酸铵稀释后经过96孔固相萃取板前处理,以含0.5%甲酸的水溶液和甲醇作为流动相,采用梯度洗脱方式在ACE Excel 3 C18-AR(2.1 mm×50 mm)色谱柱上进行分离,每个样品分析时间为4 min。在电喷雾负电离模式下,多反应监测检测离子对m/z 129.0→m/z 85.0(富马酸单甲酯)和m/z 134.0→m/z 90.0(内标,富马酸单甲酯-d5)。结果:富马酸单甲酯线性范围为4~1200 ng·mL-1(r2>0.99),定量下限为4 ng·mL-1,日内和日间精密度RSD≤9.7%,准确度相对偏差均在±4.2%以内,平均回收率RSD≤6.6%,基质效应在98.3%~100.9%之间。稳定性实验中,各种储存条件不影响对药品浓度的准确测定。结论:本方法运行时间短,灵敏度高,重复性好,适用于富马酸单甲酯血浆浓度定量检测。OBJECTIVE To establish a liquid chromatography-mass spectrometry(HPLC-MS/MS)method for the determination of monomethyl fumarate in human plasma and to apply it to the determination of monomethyl fumarate in plasma of healthy Chinese subjects.METHODS Plasma samples were diluted with ammonium acetate and then pretreated by 96-well solid phase extraction plates.The analysis was performed on an ACE Excel 3 C18-AR(2.1 mm×50 mm)chromatographic column by gradient elution using aqueous solution containing 0.5%formic acid and methanol solution as mobile phase.The analysis time of each sample was 4 min.Electrospray ionization(ESI)with a negative ionization mode and multi-reaction monitoring were adopted.The ion pairs used for quantitative analysis were m/z 129.0→m/z 85.0 for monomethyl fumarate and m/z 134.0→m/z 90.0 for monomethyl fumarate-d5.RESULTS The linear range of monomethyl fumarate was 4-1200 ng·mL-1(r2>0.99)and the lower limit of quantitation was 4 ng·mL-1 for monomethyl fumarate.The variation coefficients of intraday and interday precision were less than 9.7%,and the deviations of accuracy were within±4.2%.The RSD of recovery were less than 6.6%and the matrix effects were within 98.3%-100.9%.Various storage conditions didn’t affect the accurate determination of drug product concentration in stability experiments.CONCLUSION The method has the advantages of short run time,high sensitivity and good repeatability,and is suitable for the quantitative detection of monomethyl fumarate plasma concentration.

关 键 词：富马酸单甲酯 固相萃取法 高效液相色谱串联质谱 

分 类 号：R969.1[医药卫生—药理学]