摘要/Abstract
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
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