上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (10): 1370-.doi: 10.3969/j.issn.1674-8115.2011.10.003

• 论著（基础研究） • 上一篇    下一篇

UPLC-MS/MS测定人源Caco-2细胞中人参炔醇的含量

严忠红1, 李 林2, 汪国权3, 金玉娥3, 杨若林1, 陆 阳1   

  1. 1.上海交通大学 |医学院药学系, 上海 200025； 2.联合利华上海研发中心, 上海 200233； 3.上海市疾病预防控制中心, 上海 200336
  • 出版日期:2011-10-28 发布日期:2011-10-27
  • 通讯作者: 陆 阳, 电子信箱: huaxue@shsmu.edu.cn。
  • 作者简介:严忠红（1968—）, 女, 副教授, 博士生；电子信箱: yzh@sjtu.edu.cn。
  • 基金资助:

    上海市科委自然基金（06ZR14059, 11ZR1419000）；上海市卫生局基金（2006J006A）

UPLC-MS/MS method for determination of panaxynol in cultured Caco-2 cells

YAN Zhong-hong1, LI Lin2, WANG Guo-quan3, JIN Yu-e3, YANG Ruo-lin1, LU Yang1   

  1. 1.Department of Pharmacy, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China；2.Unilever R&D Shanghai, Shanghai 200233, China；3.Shanghai Municipal Center for Disease Control and Prevention, Shanghai 200336, China
  • Online:2011-10-28 Published:2011-10-27
  • Supported by:

    Natural Science Foundation of Shanghai, 06ZR14059, 11ZR1419000；Shanghai Municipal Health Bureau Foundation, 2006J006A

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摘要：

目的 建立一种测定人源Caco-2细胞中人参炔醇的超高效液相色谱串联质谱（UPLC-MS/MS）检测方法，分析人参炔醇在Caco-2细胞中的摄取量。方法 采用Acquity BEH C18柱（2.1 mm×100 mm，1.7 μm），流动相为水-甲醇溶液，梯度洗脱，流速为0.30 mL/min。质谱测定采用电喷雾离子源，正离子模式，多反应监测（MRM）定量离子对m/z 227→129和定性离子对m/z 227→143，测定Caco2细胞中人参炔醇的含量。结果 该方法具有良好的线性（r2 > 0.99）、精密度（≤6.2%）和准确度（－6.7%~2.1%），人参炔醇最低检出限为4 ng/mL。50 μmol/L和100 μmol/L的人参炔醇在37 ℃与细胞共同孵育2 h，Caco-2细胞中人参炔醇的摄取量分别为（20.7±1.8）nmol/mg 蛋白和（21.8±1.7）nmol/mg蛋白，差异无统计学意义（P>0.05）。结论 该方法灵敏、可靠、专属性强，可用于人参炔醇在Caco-2细胞中的吸收及转运特性研究。

关键词: 人参炔醇, 超高效液相色谱串联质谱, Caco-2

Abstract:

Objective To develop an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method to quantify the uptake of panaxynol in cultured Caco-2 cells. Methods The chromatographic separation was performed on Acquity BEH C18 column (2.1 mm×100 mm, 1.7 μm) with aqueous methanol as the mobile phase, using gradient elution at 0.3 mL/min. A triple-quadruple mass spectrometer employing electrospray ionization in positive ion mode was developed, and panaxynol in Caco-2 cells was determined using multiple reaction monitoring of precursor→product ion transitions at m/z 227→129 for quantification and m/z 227→143 for confirmation. Results The established method was validated by determining the linearity (r2>0.99), precision (≤6.2%) and accuracy (－6.7% to 2.1%). The limit of detection for panaxynol was 4 ng/mL. When incubated for 2 h at 37 ℃, the uptake was (20.7±1.8) nmol/mg protein for 50 μmol/L panaxynol and (21.8±1.7) nmol/mg protein for 100 μmol/L panaxynol, and there was no significant difference between them （P>0.05）. Conclusion This method is sensitive, reliable and specific, and can be used in determination of panaxynol in Caco-2 cells.

Key words: panaxynol, ultra-performance liquid chromatography tandem mass spectrometry, Caco-2