色谱

色谱 ›› 2013, Vol. 31 ›› Issue (3): 211-217.DOI: 10.3724/SP.J.1123.2012.10018

• 研究论文 • 上一篇    下一篇

超高效液相色谱-串联质谱法同时测定尿样中4种痕量的乌头类生物碱

张盼盼1,5, 张福成2, 王朝虹3, 蒋晔1,5*, 卢永江4   

  1. 1. 河北医科大学药学院, 河北 石家庄 050017; 2. 北京空军总医院, 北京 100142; 3. 最高人民检察院司法鉴定中心, 北京 100040; 4. 山东莒县中医院, 山东 莒县 276500; 5. 河北省法医学实验室, 河北 石家庄 050017
  • 收稿日期:2012-10-24 修回日期:2012-12-08 出版日期:2013-03-28 发布日期:2013-03-22
  • 通讯作者: 蒋晔
  • 基金资助:
    河北省卫生厅科研基金项目（08056）.

Simultaneous determination of four trace aconitum alkaloids in urine using ultra performance liquid chromatography-mass spectrometry

ZHANG Panpan1,5, ZHANG Fucheng2, WANG Zhaohong3, JIANG Ye1,5*, LU Yongjiang4   

  1. 1. School of Pharmacology, Hebei Medical University, Shijiazhuang 050017, China; 2. Air Force General Hospital, Beijing 100142, China; 3. Institute of Forensic Science of Supreme People’ s Procuratorate, Beijing 100040, China; 4. Ju County Hospital of Traditional Chinese Medicine, Ju County 276500, China; 5. Hebei Key Laboratory of Forensic Medicine, Shijiazhuang 050017, China
  • Received:2012-10-24 Revised:2012-12-08 Online:2013-03-28 Published:2013-03-22

PDF

345

被引次数

22

摘要： 建立了超高效液相色谱-串联质谱(UPLC-MS/MS)结合中空纤维微萃取(HF-LPME)同时检测尿样中痕量的乌头碱、次乌头碱、新乌头碱和滇乌头碱等4种生物碱的方法。采用HF-LPME对尿样进行提取、纯化和富集,富集倍数达102～301。同时采用电喷雾电离(ESI)、多反应监测(MRM)进行含量测定,显著提高了尿液中乌头类生物碱的检测灵敏度,4种乌头类生物碱的定量限达到0.01～0.1 ng/L,可大大延长中毒患者尿样中乌头类生物碱的检测时间窗。方法验证结果表明,尿液中乌头碱、新乌头碱和滇乌头碱在0.01～10 ng/L、次乌头碱在0.1～100 ng/L范围内线性关系良好,提取回收率为80.2%～109%,相对标准偏差小于4.6%。该方法适用于乌头类生物碱中毒案件的检测,为痕量乌头类生物碱分析提供了灵敏的分析方法。

关键词: 超高效液相色谱-质谱, 次乌头碱, 滇乌头碱, 尿样, 乌头碱, 新乌头碱, 中空纤维液相微萃取

Abstract: A rapid, specific and sensitive ultra performance liquid chromatography-mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of four trace alkaloids, including aconitine (AC), hypaconitine (HA), mesaconitine (MA) and yunaconitine (YA) in human urine samples. UPLC-MS-MS system coupled with an electrospray ionization (ESI) source was performed in multiple reaction monitoring (MRM) mode. The sample preparation was performed with hollow fiber microextraction (HF-LPME) prior to the analysis. The enrichment factors of the four alkaloids were 102-301. The separation was applied on a Waters ACQUITY UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) with a gradient elution of acetonitrile and 10 mmol/L NH4HCO3 as mobile phase. The retention times were less than 3 min. This method significantly improved the detection sensitivity, and the limits of quantitation were from 0.01 to 0.1 ng/L. The calibration curves were linear over the ranges of 0.01-10 ng/L for AC, MA and YA, 0.1-100 ng/L for HA in human urine samples, and the correlation coefficients were 0.9981, 0.9984, 0.9995 and 0.9986, respectively. The method was proved to be rapid and sensitive for aconitum alkaloid analysis in urine samples.

Key words: aconitine, hypaconitine, mesaconitine, ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), urine samples, yunaconitine, hollow fiber liquid-phase microextraction (HF-LPME)