使用LC-MS/MS方法进行生物样本分析时对溶血样本的处理策略

Strategies for dealing with hemolyzed samples in LC-MS/MS bioanalysis

在进行生物样本分析时,临床生物样本发生溶血是较常见现象。临床血浆/血清样本在发生溶血后会不同程度地改变其物理、化学性质,使得其基质与正常血浆/血清样本有所不同,对溶血样本中的目标化合物进行测定时,可能会造成浓度测定的不准确。一般而言,溶血的发生会影响待测物的色谱分离,溶血基质中可能会有一些特有的、与待测物质的共流出的内源性干扰;也可能影响待测物在质谱检测中的基质效应,甚至红细胞中释放出的酶可能影响待测物的稳定性。评价溶血基质对于生物分析的影响具有重要意义,虽然关于溶血基质效应的考察已在我国的生物分析实验室中引起了重视,但关于溶血基质效应的研究尚未达成共识。本文从生物分析行业研究的现状调查入手,探讨溶血对生物分析结果的影响以及溶血基质效应考察的规定等,结合多方面的考虑,给出了使用液相色谱-质谱联用方法进行生物样本分析时对溶血样本处理可行性的策略。

Hemolysis,which results in additional endogenous components being released from the lysed red blood cells（RBCs）,occurs frequently during the bioanalysis of clinical samples. The physical and chemical propertie of a hemolyzed clinical sample would change,which might cause additional matrix interferences. The effects of the degree of hemolysis on the method accuracy and precision should be evaluated when hemolyzed study samples were analyzed. Generally,the impact of hemolysis on bioanalysis includes：1）Chromatography Separation—additional endogenous interferences might co-elute with the target analyte in hemolyzed samples;2）Matrix Effects—the endogenous components in hemolyzed samples may cause ion suppression or enhancement in LC-MS/MS analysis. 3）Analytes Stability—analytes might not be stable in hemolyzed plasma/serum due to the enzymes released from RBCs. Therefore,it is important to evaluate the impact of hemolysis on sample analysis in LC-MS. Although the investigations of hemolysis matrix effect have drawn increasing attention in recent years,especially in cases where the sample concentrations are critical for pharmacokinetic calculation,there is no established procedure to objectively assess the risk of reporting potentially inaccurate bioanalytical results from hemolyzed samples. In this article,we investigated the proportion of hemolyzed samples in clinical samples,discussed the effects of hemolysis on determination of the concentrations of analytes. In addition,we proposed the strategy for dealing with the hemolyzed sample during LC-MS/MS bioanalysis.