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.
Abstract
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.,dopa-mine(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 mag-netic 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,re-spectively.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 rela-tive 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,in-dicating fine consistency between the two extracting methods.Conclusion The magnetic bead-based method in extracting cate-cholamine 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.
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