Quantitative analysis of ceramides by ultrahigh-performance liquid chromatography tandem mass spectrometry / 上海交通大学学报（医学版）

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 ceramides by using 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 from 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.