色谱

色谱 ›› 2011, Vol. 29 ›› Issue (01): 31-35.DOI: 10.3724/SP.J.1123.2011.00031

• 研究论文 • 上一篇    下一篇

固相萃取-高效液相色谱-串联质谱法同时测定尿液中的4种巯基尿酸

侯宏卫1*, 熊巍1, 郜娜2, 宋东奎3, 唐纲岭1, 胡清源1*   

  1. 1. 国家烟草质量监督检验中心, 河南 郑州 450001; 2. 郑州大学基础医学院临床药理研究室, 郑州大学国家药品临床研究基地, 河南 郑州 450052; 3. 郑州大学第一附属医院泌尿外科, 河南 郑州 450052
  • 收稿日期:2010-09-03 修回日期:2010-10-26 出版日期:2011-01-28 发布日期:2011-01-28
  • 通讯作者: 侯宏卫,博士,高级工程师,研究方向为烟草化学分析. Tel: (0371)67672727, E-mail: hou_hongwei@yahoo.com.cn. 胡清源,博士,研究员,研究方向为烟草化学分析. Tel: (0371)67672601, E-mail: huqy@ztri.com.cn.
  • 基金资助:

    云南中烟工业公司科技项目(No. 2010JC11).

Simultaneous determination of four mercapturic acids in human urine using solid phase extraction and liquid chromatography-tandem mass spectrometry

HOU Hongwei1*, XIONG Wei1, GAO Na2, SONG Dongkui3, TANG Gangling1, HU Qingyuan1*   

  1. 1. China National Tobacco Quality Supervision & Test Centre, Zhengzhou 450001, China; 2. National Clinical Research Base, Institute of Clinical Pharmacology, School of Medical, Zhengzhou University, Zhengzhou 450052, China; 3. Department of Urology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China
  • Received:2010-09-03 Revised:2010-10-26 Online:2011-01-28 Published:2011-01-28

PDF

809

被引次数

24

摘要： 建立了固相萃取-高效液相色谱-串联质谱(HPLC-MS/MS)同时检测人体尿液中N-乙酰基-S-(3,4-二羟基丁基)-L-半胱氨酸(DHBMA)、N-乙酰基-S-(3-羟基丙基)半胱氨酸(3-HPMA)、N-乙酰基-S-(2-氰乙基)-L-2-氨基-3-巯基羧酸(CEMA)和苯巯基尿酸(SPMA)的检测方法。冰冻的人体24 h尿液在室温下解冻,混合均匀后离心过滤,经C18固相萃取小柱净化富集后在多反应监测模式下采用HPLC-MS/MS进行定量分析。在3个添加水平下,尿液中DHBMA、3-HPMA、CEMA和SPMA的加标回收率分别为105.6%～124.4%、102.7%～106.5%、103.2%～103.9%和101.7%～104.3%,相对标准偏差为2.6%～7.7%。以不低于3倍的信噪比估算DHBMA、3-HPMA、CEMA和SPMA的检出限分别为0.062、0.031、0.020和0.003 μg/L。应用该方法检测了37例吸烟和非吸烟者的24 h尿液样本,结果发现吸烟者尿液中3-HPMA、SPMA和CEMA的平均含量比非吸烟者高3到6倍。

关键词: 固相萃取, 尿液, 巯基尿酸, 液相色谱-串联质谱

Abstract: A method was developed for the simultaneous extraction and determination of four mercapturic acids (MAs), N-acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-acetyl-S-(3-hydroxypropyl)-L-cysteine (3-HPMA), N-acetyl-S-(2-carboxyethyl)-L-cysteine (CEMA) and S-phenylmercapturic acid (SPMA), in human urine using solid phase extraction and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Frozen urine samples were thawed at room temperature, and centrifuged to remove any settled precipitate. The supernatant was then purified and concentrated by a C18 solid phase extraction column, and analyzed by HPLC-MS/MS in the multiple reaction monitoring （MRM） mode for the quantitative analysis. The ranges of recovery for DHBMA, 3-HPMA, CEMA and SPMA spiked in human urine matrix at three concentration levels were 105.6%~124.4%, 102.7%~106.5%, 103.2%~103.9% and 101.7%~104.3%, respectively, with the relative standard deviations of 2.6%~7.7%. The limits of detection (LOD, S/N≥3) were 0.062, 0.031, 0.020 and 0.003 μg/L for DHBMA, 3-HPMA, CEMA and SPMA, respectively. The method was successfully used to detect 4 MAs in 37 human urine samples from smokers and non-smokers. It was found that the contents of 3-HPMA, CEMA and SPMA in the urines from cigarette smokers were about three to six-fold more than those in the urines from the non-smokers.

Key words: mercapturic acids, solid phase extraction (SPE), urine, liquid chromatography-tandem mass spectrometry (LC-MS/MS)