›› 2019, Vol. 39 ›› Issue (12): 1353-.doi: 10.3969/j.issn.1674-8115.2019.12.003

• Original article (Basic research) • Previous Articles     Next Articles

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

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