色谱

色谱 ›› 2020, Vol. 38 ›› Issue (5): 564-571.DOI: 10.3724/SP.J.1123.2019.09018

• 研究论文 • 上一篇    下一篇

超高效液相色谱-大气压化学电离-三重四极杆质谱法同时测定血浆、尿液和瓜果类蔬菜中葫芦素B、E和I

张晓艺, 张秀尧(), 蔡欣欣, 李瑞芬   

  1. 温州市疾病预防控制中心, 浙江 温州 325001
  • 收稿日期:2019-09-17 出版日期:2020-05-08 发布日期:2020-12-10
  • 通讯作者: 张秀尧
  • 作者简介:张秀尧.Tel:057756966951, E-mail:xyzwz123@126.com

Simultaneous determination of cucurbitacin B, E and I in plasma, urine, and melon and fruit vegetables by ultra performance liquid chromatography-triple quadrupole mass spectrometry using atmospheric pressure chemical ionization

ZHANG Xiaoyi, ZHANG Xiuyao(), CAI Xinxin, LI Ruifen   

  1. Wenzhou Municipal Center for Disease Control and Prevention, Wenzhou 325001, China
  • Received:2019-09-17 Online:2020-05-08 Published:2020-12-10
  • Contact: ZHANG Xiuyao

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摘要：

建立了超高效液相色谱-三重四极杆质谱测定血浆、尿液和瓜果类蔬菜中葫芦素B、E和I的检测方法。血浆和尿液样品经固相支持液液萃取法（SLE）提取净化，瓜果类蔬菜样品经乙腈提取后用水稀释。以XBridge BEH C18色谱柱（100 mm×3.0 mm，2.5 μm）为分析柱进行分离，以甲醇-0.025%（v/v）氨水溶液为流动相进行梯度洗脱。采用大气压化学电离源，在负离子、多反应监测（MRM）模式下检测，以夹竹桃甙作为内标物，基质工作曲线内标法定量血浆和尿液中3种葫芦素；以溶剂标准曲线外标法定量瓜果类蔬菜中的待测物。血浆和尿液中3种葫芦素的检出限均为0.03 μg/L，平均加标回收率为89.0%~113%，相对标准偏差为1.7%~12.2%（n=6）；瓜果类蔬菜中3种葫芦素的检出限为5~10 μg/kg，平均加标回收率为87.6%~114%，相对标准偏差为4.1%~11.1%（n=6）。该法简单、灵敏、准确，已应用于食用苦葫芦瓜引起中毒的病人血浆和尿液，以及葫芦瓜的检测，并检出了葫芦素B。

关键词: 超高效液相色谱-三重四极杆质谱, 葫芦素B, 葫芦素E, 葫芦素I, 血浆, 尿液, 瓜果类蔬菜

Abstract:

A method for the determination of cucurbitacin B (CuB), cucurbitacin I (CuI) and cucurbitacin E (CuE) in plasma, urine and melon and fruit vegetables was developed by ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). The target analytes in plasma and urine were extracted and cleaned-up by solid supported liquid-liquid extraction, while those in melon and fruit vegetables were extracted with acetonitrile and then diluted with water. CuB, CuI and CuE were separated on an XBridge BEH C18 column (100 mm×3.0 mm, 2.5 μm) with gradient elution using mobile phases of methanol and 0.025% (v/v) ammonia aqueous solution. An atmospheric pressure chemical ionization interface was used as the ion source and the analysis was performed in negative ionization multiple reaction monitoring (MRM) mode. The cucurbitacins in plasma and urine were quantified by the matrix working standard curve internal standard method, while those in melon and fruit vegetables were quantified by the solvent standard curve external standard method. Oleandrin was used as the internal standard. The average recoveries were 89.0%-113% for the three cucurbitacins in plasma and urine, with RSDs of 1.7%-12.2% (n=6). The average recoveries were 87.6%-114% for the three cucurbitacins in melon and fruit vegetables, with RSDs of 4.1%-11.1% (n=6). The limit of detection (S/N=3) of the three cucurbitacins was 0.03 μg/L in plasma and urine, and 5-10 μg/kg in melon and fruit vegetables. The method is simple, sensitive and accurate. It has been used for the determination of cucurbitacins in bitter bottle gourd and in the plasma and urine of patients poisoned by bitter bottle gourd, CuB was successfully detected.

Key words: ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS), cucurbitacin B (CuB), cucurbitacin E (CuE), cucurbitacin I (CuI), plasma, urine, melon and fruit vegetables