液相色谱-串联质谱法监测血清茶碱浓度及室间质量评价

doi:10.3969/j.issn.1673-8640.2014.11.016

摘要/Abstract

摘要：

目的　对液相色谱-串联质谱（LC-MS/MS）检测茶碱进行方法学评价，探讨其在茶碱治疗药物监测（TDM）中的应用。方法　在血清添加放射性核素内标茶碱-D6，经蛋白沉淀稀释后采用LC-MS/MS测定。以Capcell C18 MG Ⅲ（100 mm×2.0 mm，5 μm）为分析柱进行反相色谱分离；以0.1%甲酸乙腈－0.1%甲酸水[20∶80（v/v）]为流动相，流速为0.3 mL/min；以电喷雾离子化串联四级杆质谱、正离子多反应监测进行定量检测。用建立的方法从2008年起连续参加卫生部临检中心茶碱TDM室间质量评价。结果　LC-MS/MS检测茶碱的线性范围为1～50 μg/mL，批内和批间精密度分别为2.26%~6.65%和4.70%~6.84%，准确度分别为94.14%~104.00%。单个样品的监测分析时间为3.5 min。冻融（－30 ℃↔室温反复解冻3次）、室温放置24 h、自动进样器放置24 h、长期保存（－30 ℃放置28 d）的稳定性均良好。LC-MS/MS测定结果与室间质量评价靶值偏差为2.75%，斜率为1.04，相关系数（r²）为0.983。结论　该LC-MS/MS采用放射性内标稀释，具有简单、快速、特异性和灵敏度较好的特点，连续6年测定结果符合全国室间质量评价要求，可用于茶碱的临床TDM。

关键词: 茶碱, 液相色谱-串联质谱法, 治疗药物监测, 室间质量评价

Abstract:

Objective　To perform the methodology evaluation of a liquid chromatography-tandem mass spectrometry（LC-MS/MS） for the determination of theophylline in serum， and to investigate the application significance in the therapeutic drug monitoring（TDM） for theophylline. Methods　After being added with radionuclide theophylline-D6 as internal standard， serum samples were treated with protein precipitation and determined by LC-MS/MS. A reverse phase chromatographic separation was performed on Capcell C18 MG Ⅲ analytical column（100 mm×2.0 mm， 5 μm） by using 0.1% formic acid in acetonitrile and 0.1% formic acid in water [20∶80（v/v）] as mobile phase. The flow rate was 0.3 mL/min. Theophylline and internal standard were monitored by a positive electrospray ion-tandem mass spectrometry system. The proficiency performance of LC-MS/MS was tested by participation to external quality assessment of theophylline TDM for the National Center for Clinical Laboratory. Results　The linear range of LC-MS/MS for the determination of theophylline was 1-50 μg/mL. The accuracy was 94.14%-104.00% with within-run and between-run precisions of 2.26%-6.65% and 4.70%-6.84%. The run time was 3.5 min per sample. Stabilities were good under 3 cycles of frozen-thaw for 3 times from -30 ℃ to room temperature， room temperature for 24 h， autosampler for 24 h and -30℃ for 28 d. The external quality assessment proficiency test showed the bias of 2.75%， regression slope of 1.04 and correlation coefficient（r²） of 0.983. Conclusions　The radionuclide internal standard-dilution LC-MS/MS is simple， rapid， specific and sensitive for the determination of serum theophylline. It is comparable with the external quality assessment proficiency test for consecutive 6 years， and it is suitable for theophylline TDM in clinical practice.

Key words: Theophylline, Liquid chromatography-tandem mass spectrometry, Therapeutic drug monitoring, External quality assessment

中图分类号:

R974

孙旭颖，张美微，李水军. 液相色谱-串联质谱法监测血清茶碱浓度及室间质量评价[J]. 检验医学, 2014, 29(11): 1144-1150.

SUN Xuying, ZHANG Meiwei, LI Shuijun. Liquid chromatography-tandem mass spectrometry for the monitoring of theophylline concentration in serum and its application to external quality assessment[J]. , 2014, 29(11): 1144-1150.

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