Determination of serum uric acid using high-performance liquid chromatography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method
Determination of serum uric acid using high-performance liquid chromatography (HPLC)/isotope dilution mass spectrometry (ID-MS) as a candidate reference method
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DOI：
10.1016/j.jchromb.2007.07.035
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发表时间：
2007-10-01
影响因子：
3
通讯作者：
Xu, Bei
中科院分区：
文献类型：
--
作者：
Dai, Xinhua;Fang, Xiang;Xu, Bei
尿酸是嘌呤核苷分解代谢的重要诊断指标，准确测定血清尿酸是正确诊断痛风或肾脏疾病的必要条件。描述了一种候选参考方法，包括同位素稀释与液相色谱/质谱联用（LC/MS）。将同位素标记的内标[1,3- n -15(2)]尿酸加入血清中，然后进行平衡和蛋白质去除清理，准备用于液相色谱/质谱电喷雾电离（LC/MS-ESI）分析的样品。采用LC/MS法对尿酸及其标记内标物的m/z 167和169处（m - h）（-）离子进行监测。通过将该候选参比方法在冻干人血清尿酸参比物质（标准参比物质SRM909b）上的结果与气相色谱/质谱参比方法测定的认证值以及添加尿酸的回收率研究进行比较，评估测量的准确性。该方法与离子交换衍生化同位素稀释（ID）气相色谱-质谱法（ID- gc /MS）相比较，具有良好的性能。该方法测定的尿酸值与检定值吻合较好，误差在0.10%以内。尿酸的回收和添加量在三种浓度下是一致的。本方法用于冷冻血清池中尿酸的测定。LC/MS分析的组内CVs值为0.08 ~ 0.18%，组间CVs值为0.02 ~ 0.07%，精密度较高。液相色谱/串联质谱电喷雾电离（LC/MS/MS- esi）分析。LC/MS和LC/MS/MS结果一致性很好（在0.14%以内）。该LC/MS方法具有良好的准确度和精密度，且分析速度快，不需要衍生化阶段，可作为候选参考方法。这种方法可以作为一种替代的参考方法，为常规方法提供一个可以比较的精度基础。(c) 2007 Elsevier B.V.版权所有。
Uric acid is an important diagnostic marker of catabolism of the purine nucleosides, and accurate measurements of serum uric acid are necessary for proper diagnosis of gout or renal disease appearance. A candidate reference method involving isotope dilution coupled with liquid chromatography/mass spectrometry (LC/MS) has been described. An isotopically labeled internal standard, [1,3-N-15(2)] uric acid, was added to serum, followed by equilibration and protein removal clean up to prepare samples for liquid chromatography/mass spectrometry electrospray ionization (LC/MS-ESI) analyses. (M-H)(-) ions at m/z 167 and 169 for uric acid and its labeled internal standard were monitored for LC/MS. The accuracy of the measurement was evaluated by a comparison of results of this candidate reference method on lyophilized human serum reference materials for uric acid (Standard Reference Materials SRM909b) with the certified values determined by gas chromatography/mass spectrometry reference methods and by a recovery study for the added uric acid. The method performed well against the established reference method of ion-exchange followed by derivatization isotope dilution (ID) gas chromatography mass spectrometry (ID-GC/MS). The results of this method for uric acid agreed well with the certified values and were within 0.10%. The amounts of uric acid recovered and added were in good agreement for the three concentrations. This method was applied to determine uric acid in samples of frozen serum pools. Excellent precision was obtained with within-set CVs of 0.08-0.18% and between-set CVs of 0.02-0.07% for LC/MS analyses. Liquid chromatography/tandem mass spectrometry electrospray ionization (LC/MS/MS-ESI) analysis was also performed. The LC/MS and LC/MS/MS results were in very good agreement (within 0.14%). This LC/MS method, which demonstrates good accuracy and precision, and is in the speed of analysis without the need for a derivatization stage, qualifies as a candidate reference method. This method can be used as an alternative reference method to provide an accuracy base to which the routine methods can be compared. (c) 2007 Elsevier B.V. All fights reserved.