高等学校化学学报

高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (12): 2716.doi: 10.7503/cjcu20130704

• 分析化学 • 上一篇    下一篇

UPLC-MS/MS法测定高尿酸血症大鼠血清及尿液中的尿酸和肌酐

马青山1, 王茜2, 赵凯姝1, 翟淑波1, 刘舒3, 刘志强3   

  1. 1. 吉林大学第一医院小儿肾病科, 长春 130021;
    2. 吉林大学中日联谊医院肾内科, 长春 130033;
    3. 中国科学院长春应用化学研究所, 长春 130022
  • 收稿日期:2013-07-25 出版日期:2013-12-10 发布日期:2013-10-11
  • 作者简介:刘志强，男，博士，研究员，博士生导师，从事天然产物质谱及中药活性成分筛选研究. E-mail：liuzq@ciac.jl.cn
  • 基金资助:

    国家自然科学基金（批准号：21175128，21073178，81303280）和吉林省科技发展重点项目（批准号：20110928）资助.

UPLC-MS/MS Method for Determination of Uric Acid and Creatinine in Serum and Urine of Hyperuricemic Mice

MA Qing-Shan1, WANG Qian2, ZHAO Kai-Shu1, ZHAI Shu-Bo1, LIU Shu3, LIU Zhi-Qiang3   

  1. 1. Department of Pediatric Nephrology, The First Hospital of Jilin University, Changchun 130021, China;
    2. Department of Nephrology, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
    3. Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China
  • Received:2013-07-25 Online:2013-12-10 Published:2013-10-11

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被引次数

15

摘要：

建立了一种测定血清和尿液中尿酸和肌酐含量的超高效液相色谱-串联质谱联用（UPLC-MS/MS）方法. 样品经自动进样器引入液相色谱通过预柱富集和去除杂质后，直接引入质谱中进行分析. 采用正离子电喷雾电离模式下的多反应监控模式对肌酐进行定量分析，纯溶剂标准曲线的肌酐线性范围为0.03~400 μmol/L；基质标准曲线的肌酐线性范围为0.2~400 μmol/L；它们的最低定量限分别为0.03和0.2 μmol/L. 采用负离子电喷雾电离模式下的多反应监控模式对尿酸进行定量分析，纯溶剂标准曲线的尿酸线性范围为0.1~350 μmol/L；基质标准曲线的尿酸线性范围为0.5~300 μmol/L；它们的最低定量限分别为0.1和0.5 μmol/L. 该方法的提取回收率在91.8%~103.7%之间，日内和日间RSD分别小于5.9%和6.8%，满足生物分析的要求. 利用该方法进行抗痛风中药筛选及其作用机制研究，结果表明，与阳性对照药物别嘌呤醇和苯溴马隆相比，中药二妙丸、黄柏和苍术均能在一定程度上降低血尿酸水平，并可显著逆转肾功能损伤，对肾功能具有一定的保护作用.

关键词: 高尿酸血症, 尿酸, 肌酐, 三重四极杆质谱

Abstract:

An ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS) method for the determination of uric acid and creatinine in serum and urine was established and applied to screening of anti-gout medicines. The automatic sampler and the pump of an LC apparatus were used to inject sample and mobile phase into a chromatography guard column which was used to remove most interference components and enrich the target compounds. MS/MS analysis was carried out in positive ion mode and in multiple reaction monitoring(MRM) mode for creatinine, linearity of the MS/MS assay without and with matrix were 0.03—400 μmol/L and 0.2—400 μmol/L of creatinine, with a limit of detection(LOD) of 0.03 and 0.2 μmol/L, respectively. The uric acid was analyzed in negative ion mode, linearity of the MS/MS assay without and with matrix were 0.1—350 μmol/L and 0.5—300 μmol/L of uric acid, with a LOD of 0.1 and 0.5 μmol/L, respectively. An analytical recovery of this method was 91.8%—103.7% and the RSDs for intra-day and inter-day were less than 5.9% and 6.8% respectively. The results meet the requirements for a bioanalytical assay. Subsequently, the established method was applied to screening of anti-gout medicines. The results showed that ermiaowan, Phellodendri Chinensis Cortex and Atractylodis Rhizoma were able to reduce blood uric acid levels at some degree compared with the positive control drugs, including allopurinol and benzbromarone. In addition, they can significantly reverse the renal damage and protect the renal function.

Key words: Hyperuricemia, Uric acid, Creatinine, Triple quadrupole mass spectrometry

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