检验医学 ›› 2015, Vol. 30 ›› Issue (5): 422-426.DOI: 10.3969/j.issn.1673-8640.2015.05.004

• 质谱技术在临床生化检验中的应用专题 • 上一篇    下一篇

液相色谱-串联质谱法测定尿酸及与常规检测方法的比较

张海晨1, 李水军2, 孙贺伟3, 宋云霄1   

  1. 1. 上海市徐汇区中心医院(中国科学院上海临床研究中心)检验科,上海 200031
    2. 上海市徐汇区中心医院(中国科学院上海临床研究中心)中心实验室,上海 200031
    3. 上海大学生命科学学院,上海 200444
  • 收稿日期:2015-01-21 出版日期:2015-05-30 发布日期:2015-06-17
  • 作者简介:null

    作者简介:张海晨,男,1973年生,副主任技师,主要从事临床生物化学检验工作。

    通讯作者:李水军,联系电话:021-54031835。

  • 基金资助:
    上海市徐汇区卫生与计划生育委员会科研基金资助项目(SHXH201301)

Liquid chromatography-tandem mass spectrometry for uric acid and its comparison with clinical routine determination method

ZHANG Haichen1, LI Shuijun2, SUN Hewei3, SONG Yunxiao1   

  1. 1. Department of Clinical Laboratory, Shanghai Xuhui Central Hospital (Shanghai Clinical Research Center, Chinese Academy of Sciences), Shanghai 200031, China
    2. Central Laboratory, Shanghai Xuhui Central Hospital (Shanghai Clinical Research Center, Chinese Academy of Sciences), Shanghai 200031, China
    3. Biological College, Shanghai University, Shanghai 200444, China
  • Received:2015-01-21 Online:2015-05-30 Published:2015-06-17

摘要: 目的 建立液相色谱-串联质谱(LC-MS/MS)检测血清尿酸的方法,并与临床常规生化方法进行比较,为临床实验室不同检测系统尿酸检测结果的一致性提供参考。方法 血清添加同位素内标尿酸-15N2,经乙腈处理吹干重组后采用LC-MS/MS测定。以Capcell C18 MGⅢ为分析柱进行反相色谱分离,以5 mmol/L乙酸铵+0.1%乙酸水溶液和甲醇(90∶10,v/v)为流动相,等度洗脱,流速0.3 mL/min,电喷雾离子化串联四极杆质谱,负离子多反应监测,尿酸离子通道为167/124 amu(定量)、167/96 amu(定性);尿酸-15N2离子通道为169/125 amu。LC-MS/MS进行方法学评价后与临床尿酸常规检测方法(酶紫外法和酶比色法)进行比较。结果 LC-MS/MS检测尿酸的线性范围为30~952 μmol/L,批内和批间精密度分别为2.01%~6.23%和4.55%~8.08%,准确度为96.5%~103.4%。尿酸的色谱保留时间为1.5 min,单份样品的分析时间为3 min。酶紫外法、酶比色法与LC-MS/MS的平均偏差分别为-10.02%、-9.88%;线性相关方程分别为Y酶紫外法=0.898XLC-MS/MS+2.15,r=0.978;Y酶比色法=0.845XLC-MS/MS+22.15,r=0.983。LC-MS/MS与美国标准技术研究所(NIST)定值参考物质的平均偏差为1.56%±0.65%,与卫生部临床检验中心2014年代谢物正确度验证样品的定值的偏差为-0.34%~3.05%。结论 采用LC-MS/MS可简便、准确地定量检测尿酸。酶紫外法、酶比色法的血清尿酸测定结果与LC-MS/MS具有较好的可比性。

关键词: 尿酸, 液相色谱-串联质谱法, 尿酸酶紫外法, 尿酸酶比色法

Abstract: Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)for serum uric acid, and to compare LC-MS/MS with clinical routine determination methods. Methods Uric acid-15N2 was added as internal standard, serum samples were precipitated with acetonitrile and dried with nitrogen flow. A reversed-phase chromatographic separation was performed on Capcell C18 MGⅢ analytical column by using 5 mmol/L ammonium acetate plus 0.1% acetate acid in water and methanol(90∶10,v/v)as mobile phase. The flow rate was 0.3 mL/min. Uric acid and internal standard were monitored by a negative electrospray ion-tandem mass spectrometry system using ion transitions of 167/124 amu (quantitation) and 167/96 amu (qualitation) for uric acid and 169/125 amu for uric acid-15N2. After being validated, the LC-MS/MS was compared with the uricase ultraviolet (UV) method and uricase colorimetric (UC) method. Results The LC-MS/MS was validated over a concentration range of 30-952 μmol/L. Within-run and between-run precisions were 2.01%-6.23% and 4.55%-8.08%, respectively. The accuracy was 96.5%-103.4%. The retention time of uric acid was 1.5 min, and total run time was 3 min. The linear correlation formulas among LC-MS/MS, uricase UV method and uricase UC method were YUV=0.898XLC-MS/MS +2.15, r=0.978 and YUC=0.845XLC-MS/MS+22.15, r=0.983. The average biases were 1.56%±0.65% between LC-MS/MS and the National Institute of Standards and Technology (NIST) standard reference material and -0.34%-3.05% between LC-MS/MS and correctness verification samples, 2014 from the National Center for Clinical Laboratory. Conclusions Serum uric acid can be simply and accurately measured by LC-MS/MS. There is good comparability between LC-MS/MS, uricase UV method and uricase UC method.

Key words: Uric acid, Liquid chromatography-tandem mass spectrometry method, Uricase ultraviolet method, Uricase colorimetric method

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