上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (12): 1353-.doi: 10.3969/j.issn.1674-8115.2019.12.003

• 论著·基础研究 • 上一篇    下一篇

基于超高效液相色谱-质谱联用技术的神经酰胺定量分析

黄庆霞1, 2，雷和花1，唐惠儒3，王玉兰4   

  1. 1. 中国科学院武汉物理与数学研究所波谱与原子分子物理国家重点实验室，武汉磁共振中心，武汉430071；2．中国科学院大学，北京100049；3．复旦大学附属中山医院，生命科学院，遗传工程国家重点实验室，人类表型组研究院，代谢组学与系统生物学实验室，上海200433；4．新加坡南洋理工大学李光前医学院，新加坡表型中心，生命科学院，新加坡639646
  • 出版日期:2019-12-28 发布日期:2020-02-06
  • 通讯作者: 王玉兰，电子信箱：yulan.wang@ntu.edu.sg。
  • 作者简介:黄庆霞（1990—），女，博士生；电子信箱：qiangxhuang@126.com。
  • 基金资助:
    国家重点研发计划（2017YFC0906800）

Quantitative analysis of ceramidesultrahigh-performance liquid chromatography tandem mass spectrometry

HUANG Qing-xia, LEI He-hua, TANG Hui-ru, WANG Yu-lan   

  1. 1. State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan 430071, China; 2. University of Chinese Academy of Sciences, Beijing 100049, China; 3. State Key Laboratory of Genetic Engineering, Zhongshan Hospital and School of Life Sciences, Laboratory of Metabonomics and Systems Biology, Human Phenome Institute, Fudan University, Shanghai 200433, China; 4. Singapore Phenome Center, Lee Kong Chian School of Medicine, School of Biological Sciences, Nanyang Technological University, Singapore 639646, Singapore
  • Online:2019-12-28 Published:2020-02-06
  • Supported by:
    National Key Research and Development Program，2017YFC0906800

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摘要： 目的·研发一种具有高通量、高灵敏度和准确度的神经酰胺定量分析方法。方法·应用超高效液相色谱串联三重四极杆质谱（ultrahigh-performance liquid chromatography tandem triple-quadrupole mass spectrometry，UHPLC-MS）对14种神经酰胺进行定量分析，使用的色谱柱为ZORBAX Eclipse plus C8 (2.1 mm×150 mm，1.8 μm)，流动相A为超纯水，流动相B为甲醇，流动相A和B中均加入2.5 mmol/L乙酸铵。采用多反应监测模式和正离子模式分析神经酰胺。结果·该方法能在10 min内定量分析14种神经酰胺，检测限（limit of detection，LOD）和定量限（limit of quantification，LOQ）分别为0.06～0.42 nmol/L和0.17～1.26 nmol/L。准确度范围为73.02%～122.39%，保留时间和积分面积的相对标准偏差分别为0.07%～0.54%和2.26%～13.98%。结论·基于UHPLC-MS技术的神经酰胺定量分析方法灵敏度高、重现性好、分析效率高，可用于检测细胞等生物样品中的神经酰胺含量。

关键词: 神经酰胺, 超高效液相色谱, 三重四极杆质谱, 定量分析

Abstract:

Objective · To develop a method that can quantitatively analyze a wide range of ceramides with high throughout, sensitivity and accuracy. Methods · Ultrahigh-performance liquid chromatography tandem triple-quadrupole mass spectrometry (UHPLC-MS) was employed for quantitative analysis of 14 ceramidesusing ZORBAX Eclipse plus C8 (2.1 mm×150 mm, 1.8 μm) column. Solvent A consisted of water, solvent B consisted of methanol, and 2.5 mmol/L ammonium acetate was added to both. Multiple reaction monitoring (MRM) mode and positive ion mode were used to analyze the ceramides. Results · This method can quantitatively analyze 14 ceramides in 10 minutes with the limit of detection (LOD) and limit of quantification (LOQ) between 0.06-0.42 nmol/L and 0.17-1.26 nmol/L, respectively. The accuracy ranged 73.02% to 122.39%, and the relative standard deviation (RSD) of retention time and peak area were between 0.07%-0.54% and 2.26%-13.98%, respectively. Conclusion · Quantitative analysis of ceramides based on UHPLC-MS has high sensitivity, reproducibility and efficiency, which can be used to analyze the ceramides in biological samples such as cells.

Key words: ceramide, ultrahigh-performance liquid chromatography, triple-quadrupole mass spectrometry, quantitative analysis