同位素稀释液相色谱串联质谱法测定血清葡萄糖  

Determination of serum glucose by isotope dilution liquid chromatography tandem mass spectrometry

作　　者：张天娇[1] 张传宝[1] 张江涛[1] 周伟燕[1] 赵海舰[1] 闫颖[1] 胡翠华[1] 王冬环[1] 汪静[1] 谢洁红[1] 马嵘[1] 申子瑜[1] 陈文祥[1] 

机构地区：[1]卫生部北京医院卫生部临床检验中心,100730

出　　处：《中华检验医学杂志》2010年第1期75-80,共6页Chinese Journal of Laboratory Medicine

基　　金：国家科技支撑计划资助项目（2007BA105809）;国家高技术研究发展计划（863计划）资助项目（2006AA020909）

摘　　要：目的建立一种使用同位素稀释液相色谱串联质谱法（ID—LC／MS／MS）测定血清葡萄糖的候选参考方法。方法以[^13C6]葡萄糖为内标，用重量法准确地与血清混合，除去蛋白后在碱性条件下与1-苯基-3-甲基-5-吡唑酮反应，用LC／MS／MS测定葡萄糖和内标衍生产物，以包括法定量。结果血清葡萄糖测定的批内、批问和总变异系数的平均值分别为0．36％（范围0．28％-0．42％）、0．47％（范围0．20％～0．67％）和0．61％（范围0．42％～0．76％）。加样回收试验的回收率范围为99．0％-100．9％。分析参考物质SRM965a，测定结果与认定值的平均偏差为-0．20％（范围-0．39％～＋0．11％）。结论建立了ID—LC／MS／MS法测定血清葡萄糖的方法，方法准确、精密、简便，可望作为血清葡萄糖测定的参考方法。Objective To develop a candidate reference method for the measurement of serum glucose based on isotope dilution hquid chromatography tandem mass spectrometry （ID-LC/MS/MS）. Methods An internal standard [^13 C6 ] glucose was added to serum samples and equilibrated with endogenous glucose. Serum proteins were removed by a precipitation with anhydrous ethanol. Serum glucose and the internal standard were then reacted with 1-phenyl-3-methyl-5-pyrazolone and the formed derivatives were analyzed by liquid chromatography tandem mass spectrometry with multiple reaction monitoring （MRM） The method was calibrated with bracketing calibrators and serum glucose concentrations were calculated by comparing the peak area ratios of samples with that of the calibrators. Results The within-run, between-run and total coefficients of variation averaged 0. 36% , 0. 47% and 0. 61% , respectively. The analytical recoveries ranged from 99.0% to 100. 9%. Results of analyzing the certified reference material SRM 965a showed an average biases of -0. 20%. Conclusions An ID-LC/MS/MS method for measuring serum glucose has been developed. The method is highly precise and accurate and may be used as a candidate reference method.

关 键 词：血糖 同位素标记 色谱法 液相 串联质谱法 

分 类 号：R446.1[医药卫生—诊断学]