不同品牌蛋白沉淀板对人血浆中霉酚酸含量测定结果比较

Comparative study on content determination of mycophenolic acid in human plasma by two brands of protein precipitation filter plates

目的:利用超高效液相色谱-串联质谱法,比较不同品牌(赛默飞和岛津)的蛋白沉淀板测定人血浆霉酚酸的效果。方法:含霉酚酸的血浆样本经分别2种蛋白沉淀板进行前处理,以霉酚酸-d3为内标,采用Acquity UPLC BEH-C_(18)柱(2.1 mm×50 mm, 1.7μm)分离,流动相包括水(含0.1%甲酸,2 mmol·L^(-1)醋酸铵,V/V)和乙腈(含0.1%甲酸,V/V),梯度洗脱,流速0.2 mL·min^(-1)。正离子检测模式(+ESI)下,采用多离子反应监测(MRM)模式进行分析,霉酚酸的离子对为m/z 321.2-207.1,内标的离子对为324.0-210.0。比较各蛋白沉淀板的准确度、精密度、基质效应、提取回收率等指标。结果:霉酚酸的线性范围为0.312 5~20μg·mL^(-1),定量下限为0.312 5μg·mL^(-1)。各品牌的蛋白沉淀板所获得的准确度、精密度、基质效应和提取回收率均符合要求。结论:这2种蛋白沉淀板操作简便、快速、准确、且精密度良好,方法学考察结果没有显著性差异,均适用于人血浆中霉酚酸的分析,可用于临床治疗药物监测。

OBJECTIVE To compare two brands(Thermo Fisher vs. Shimadzu)of protein-precipitation filter plates on the content determination of mycophenolic acid in human plasma by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). METHODS The plasma samples were pretreated by acetonitrile and processed by two brands of protein-precipitation filter plates respectively.Mycophenolic acid-d3 was employed as an internal standard.Then analytes were gradient eluted on a UPLC? BEH-C_(18) column(2.1 mm×50 mm, 1.7 μm)by UPLC-MS/MS with positive ionization.Ions monitored in multiple reaction monitoring(MRM)mode were m/z 321.2-207.1 for mycophenolic acid and 324.0-210.0 for mycophenolic acid-d3(internal standard)respectively.RESULTS Excellent liner relationship was obtained from the range of 0.312 5-20 μg·mL^(-1) and the lower quantitative limit of mycophenolic acid was 0.312 5 μg·mL^(-1).Both protein-precipitation filter plates fulfilled the validation acceptance criteria of accuracy, extraction recovery, matrix effect, intra-assay and inter-assay precision.CONCLUSION These two brands of protein-precipitation filter plates are simple, rapid and accurate with excellent precision, showing equivalent quantitative performance for monitoring mycophenolic acid.And both brands are to be applied for routine clinical monitoring of drug concentrations.