色谱

色谱 ›› 2013, Vol. 31 ›› Issue (11): 1102-1105.DOI: 10.3724/SP.J.1123.2013.07033

• 技术与应用 • 上一篇    下一篇

高效液相色谱法同时测定尿液中4种非蛋白氮的含量

马玉花1, 黄冬群1, 张瑞1, 徐世茹2, 封顺1   

  1. 1. 新疆大学石油天然气精细化工教育部自治区重点实验室, 新疆 乌鲁木齐 830046;
    2. 新疆维吾尔自治区人民医院肾病科, 新疆 乌鲁木齐 830001
  • 收稿日期:2013-07-22 修回日期:2013-08-19 出版日期:2013-11-28 发布日期:2013-12-05
  • 通讯作者: 封顺
  • 基金资助:

    国家自然科学基金项目(21075105);新疆维吾尔自治区教育厅重点项目(XJEDU2010I03).

Simultaneous determination of four common nonprotein nitrogen substances in urine by high performance liquid chromatography

MA Yuhua1, HUANG Dongqun1, ZHANG Rui1, XU Shiru2, FENG Shun1   

  1. 1. Key Laboratory of Oil & Gas Fine Chemicals, Ministry of Education, Xinjiang University, Urumqi 830046, China;
    2. Department of Nephrology, Xinjiang Uygur Autonomous Region People's Hospital, Urumqi 830001, China
  • Received:2013-07-22 Revised:2013-08-19 Online:2013-11-28 Published:2013-12-05

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412

被引次数

3

摘要：

建立了高效液相色谱(HPLC)同时测定尿液中4种常见的非蛋白氮物质——肌酸(Cr)、尿肌酐(Cn)、尿酸(Ua)及假尿嘧啶核苷(Pu)含量的方法。尿液首先经丙酮沉淀法除去大分子化合物,然后冷冻干燥,复溶后直接进行HPLC分析。色谱分离选用Waters RP18 Column (150 mm×4.60 mm,3.5 μm),以10.0 mmol/L磷酸缓冲溶液 (pH 4.78)和乙腈为流动相进行梯度洗脱,流速为0.8 mL/min,于220 nm波长下检测,可在7 min内完成4种非蛋白氮的快速分离分析。结果表明Cr、Cn、Ua、Pu在0.1~250 mg/L范围内线性关系均良好(相关系数均大于0.999),检出限分别为9.31、26.19、4.70、6.30 μg/L,加标回收率为81%~111%,峰高的相对标准偏差(RSD, n=3)为0.23%~2.78%,满足定量要求。该法简便、快速,结果准确可靠,为2型糖尿病(T2DM)患者肾功能损坏的研究提供了检测手段。

关键词: 高效液相色谱法, 肌酸, 假尿嘧啶核苷, 尿肌酐, 尿酸, 尿液

Abstract:

A high performance liquid chromatographic (HPLC) method was proposed to simultaneously determine four common nonprotein nitrogen substances, including creatine (Cr), creatinine (Cn), uric acid (Ua) and pseudouridine (Pu) in urine. After proteins being removed by acetone precipitation method, freeze drying and redissolving, the urine samples were analyzed by HPLC. Chromatographic separation was performed on a Waters RP18 Column (150 mm×4.60 mm, 3.5 μm) in gradient elution mode using 10.0 mmol/L KH2PO4 solution (pH 4.78) and acetonitrile as mobile phases at a flow rate of 0.8 mL/min. The samples were detected at 220 nm. Rapid separation was achieved within 7 min. Under the optimized conditions, good linearities of four common nonprotein nitrogen substances were obtained in the range of 0.1-250 mg/L. The detection limits were 9.31 (Cr), 26.19 (Cn), 4.70 (Ua), and 6.30 (Pu) μg/L and the recoveries were in the range of 81%-111% with the relative standard deviations of 0.23%-2.78% (n=3). The results demonstrate that this method is simple, rapid and accurate with good reproducibility, and can provide early diagnosis and preliminary judgment for type 2 diabetes mellitus (T2DM) patients with renal damage.

Key words: creatine, creatinine, high performance liquid chromatography (HPLC), pseudouridine, uric acid, urine

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