同位素稀释液相色谱串联质谱法测定人血清尿酸  

Determination of serum uric acid by isotope dilution liquid chromatography tandem mass spectrometry

作　　者：张传宝[1] 张江涛[1] 张天娇[1] 周伟燕[1] 陈文祥[1] 申子瑜[1] 王冬环[1] 

机构地区：[1]卫生部北京医院卫生部临床检验中心,北京100730

出　　处：《检验医学》2009年第12期878-882,共5页Laboratory Medicine

基　　金：国家科技支撑计划资助项目(2007BAI05B09);国家高技术研究发展计划(863)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（ID-LC/MS/MS）测定人血清尿酸的候选参考方法。方法用[1,3]1-5N2尿酸作内标,与一定量的血清充分混匀,加入等量乙腈沉淀血清蛋白并提取尿酸,氯仿纯化上清液后用液相色谱串联质谱（LC/MS/MS）分离测定。优化LC/MS/MS的色谱条件,评价该法的精密度、回收率和偏倚,并计算不确定度。结果对3个浓度水平的室间质量评价用质控血清测定3批,批内、批间和总变异系数（CV）的均值（范围）分别为0.54%（0.44%-0.69%）、0.66%（0.45%-0.79%）和0.86%（0.66%-1.02%）。加样回收试验回收率范围为98.38%-102.64%。美国国家标准与技术研究所（NIST）标准物质SRM 909b-Ⅰ、Ⅱ的测定均值与靶值的偏倚分别为0.04%和0.25%。3种质控血清的相对扩展不确定度分别为1.80%、2.19%和2.04%。结论建立的ID-LC/MS/MS测定血清尿酸的方法精密、准确,有望作为血清尿酸测定的参考方法。Objective To develop a candidate reference method for the measurement of serum uric acid by isotope dilution liquid chromatography tandem mass spectrometry（ID-LC/MS/MS）.Methods An isotope labeled internal standard（IS） -15N2 uric acid was weighed and spiked to serum samples.Acetonitrile was applied to precipitate serum proteins and extract uric acid and IS.After being treated with chloroform for further clean-up,the samples were analyzed by LC/MS/MS.Serum uric acid was quantified by a bracketing calibration.Results The within-run,between-run and total coefficients of variation（CV）of 3 batches of measurements of 3 levels of lyophilized sera used in external quality assessment ranged from 0.44% to 0.69%,0.45 % to 0.79% and 0.66% to 1.02%,and the averages were 0.54%,0.66% and 0.86%,respectively.The analytical recoveries ranged from 98.38% to 102.64% with an average of 100.55%.The results of analyzing the certified reference material SRM909b-Ⅰand Ⅱ showed 0.04% and 0.25% proportional bias compared with certified values.Evaluated uncertainty of measurement for the determination of 3 levels of lyophilized sera and values of the relative expanded uncertainty were 1.80%,2.19% and 2.04%.Conclusions An ID-LC/MS/MS method for measuring serum uric acid has been developed.The method is highly precise and accurate and may be used as a candidate reference method for serum uric acid measurement.

关 键 词：尿酸 同位素标记 液相色谱 串联质谱法 

分 类 号：R446.1[医药卫生—诊断学]