磁珠法结合液相色谱串联质谱技术应用于人血浆儿茶酚胺检测  

Application of magnetic bead-based liquid chromatography-tandem mass spectrometry(LC-MS/MS)for detection of catecholamines in human plasma

作　　者：花蕾 韩静贤[1] 韩文超 张思琪 HUA Lei;HAN Jingxian;HAN Wenchao;ZHANG Siqi(Henan Key Laboratory of Cardiac Remodeling and Transplantation,the Seventh People′s Hospital of Zhengzhou,Zhengzhou 450016,Henan,China;Department of Pharmacy,the Seventh People′s Hospital of Zhengzhou,Zhengzhou 450016,Henan,China)

机构地区：[1]郑州市第七人民医院河南省心脏重构与移植重点实验室,郑州450016 [2]郑州市第七人民医院药学部,郑州450016

出　　处：《临床检验杂志》2024年第12期881-886,共6页Chinese Journal of Clinical Laboratory Science

基　　金：河南省医学科技攻关项目(LHGJ20240900);郑州市医疗卫生领域科技创新指导计划(2024YLZDJH232)。

摘　　要：目的评估利用磁珠法结合液相色谱串联质谱(LC-MS/MS)技术检测血浆中儿茶酚胺代谢物的性能。方法利用磁珠法结合LC-MS/MS检测血浆中儿茶酚胺的代谢物:多巴胺(DA)、去甲肾上腺素(NE)、肾上腺素(E)、3-甲氧基酪胺(3-MT)、甲氧基去甲肾上腺素(NMN)和甲氧基肾上腺素(MN),并通过测定线性范围、检测限、精密度、基质效应等指标对方法性能进行验证。收集81份临床血浆样本,分别采用磁珠法和传统固相萃取法(SPE)进行前处理,经LC-MS/MS测定后,比较2种前处理方法结果的一致性,并进行统计学分析。结果磁珠法提取DA、NE、E、3-MT、NMN和MN的线性相关系数均大于0.99,定量限分别为10.41、80.2、4.55、3.89、11.2、1.12 pg/mL,检测限分别为2.75、26.5、1.5、1.35、3.1、0.34 pg/mL,日内和日间精密度以变异系数(CV)表示均不超过15%,相对回收率分别为100.54%~102.21%、95.78%~101.85%、95.56%~99.09%、95%~102.72%、94.97%~100.19%、94.77%~100.65%,相对偏差均在±15%以内。相对基质效应均在85%~115%范围内。磁珠法与SPE法提取DA、NE、E、3-MT、NMN和MN的结果比对,相对百分偏差分别为0.93%、0.99%、0.95%、0.92%、0.97%、1.02%,表明2种方法具有良好的一致性。结论磁珠法提取血浆中儿茶酚胺代谢物的分析性能良好,操作快速简便,有助于提高临床检测的自动化程度和通量,具有良好的应用前景。Objective To evaluate the performance of magnetic bead-based liquid chromatography-tandem mass spectrometry(LC-MS/MS)in extracting and detecting catecholamine metabolites in plasma.Methods The catecholamine metabolites,i.e.,dopamine(DA),norepinephrine(NE),epinephrine(E),3-methoxytyramine(3-MT),normetanephrine(NMN),and metanephrine(MN),were extracted from plasma using magnetic bead-based LC-MS/MS.The performance of the method was validated by assessing linear range,detection limit,precision,and matrix effects.Clinical plasma samples(n=81)were collected,and the results of magnetic bead-based extraction were compared with traditional solid-phase extraction(SPE)using LC-MS/MS and the statistical analysis were conducted for consistency of the two extraction methods.Results The linear correlation coefficients of DA,NE,E,3-MT,NMN and MN extracted using magnetic beads were all greater than 0.99,and the limits of quantification were determined to be 10.41,80.2,4.55,3.89,11.2 and 1.12 pg/mL,and the limits of detection were established as 2.75,26.5,1.5,1.35,3.1 and 0.34 pg/mL,respectively.The intra-and inter-day precision expressed as coefficient of variation(CV)were both within 15%.The relative recovery rates ranged from 100.54%to 102.21%for DA,95.78%to 101.85%for NE,95.56%to 99.09%for E,95%to 102.72%for 3-MT,94.97%to 100.19%for NMN,and 94.77%to 100.65%for MN with the range of all the relative deviations within±15%.All the relative matrix effects were within the range of 85%to 115%.The comparisons of magnetic bead extraction with SPE for DA,NE,E,3-MT,NMN and MN showed the relative percent deviations of 0.93%,0.99%,0.95%,0.92%,0.97%and 1.02%,respectively,indicating fine consistency between the two extracting methods.Conclusion The magnetic bead-based method in extracting catecholamine metabolites from plasma was demonstrated to possess excellent analytical performance,rapid and simple operation,and could enhance the automation degree and throughput in the process of clinical testing,showing promising application prospects.

关 键 词：液相色谱串联质谱 儿茶酚胺代谢物 磁珠法 固相萃取法 

分 类 号：R446[医药卫生—诊断学]