基于高效液相色谱-串联质谱的磷脂酰胆碱的分离与表征

Separation and characterization of phosphatidylcholine based on high performance liquid chromatography-tandem mass spectrometry

磷脂酰胆碱是维持细胞正常运作、调节生物膜功能和人体正常代谢的重要物质。其结构多变且相似度高,拥有相当多的同系物,导致磷脂酰胆碱化合物的分离与鉴定一直是一个难点问题。基于高效液相色谱-串联质谱方法,以前期制备得到的磷脂酰胆碱产品为研究对象,对其组成成分进行了分离和结构表征。首先通过优化固定相和流动相建立了反相色谱分离的方法:Symmetry C18为固定相,0.2%氨水的甲醇/乙腈为流动相。然后采用HPLC-MS/MS方法对其中主要的磷脂酰胆碱进行分离和表征。磷脂酰胆碱类化合物的分子离子峰为偶数,在正离子模式下产生特征碎片离子m/z 184。负离子模式下产生[R_2COO]^-和[R_1COO]^-特征碎片离子,可用来判断脂肪酸的种类,产生的[M-CH3-R_2CO]^-离子可用于判断脂肪酸的连接位点。最终,检测到样品中含有12种磷脂酰胆碱类化合物。

Phosphatidylcholine is an important material to keep normal operation of the cell,regulate the biological function of membrane and maintain normal metabolism of human body.Its structure is complex and high similarity with considerable homologues,leading to difficult isolation and identification.Based on the method of high performance liquid chromatography tandem mass spectrometry,the phosphatidylcholine of a previous prepared sample were separated and characterized.Firstly,by optimizing the stationary phase and mobile phase,a reversed-phase chromatography method was established:symmetry C18 was the stationary phase,and 0.2%methanol/acetonitrile was used as mobile phase.Then,the HPLC-MS/MS method was used to isolate and characterize the phosphatidylcholine.In positive mode,the molecular ion peak of phosphatidylcholine was even,producing characteristic fragment ions of m/z 184.In negative mode,fragment ions of[R 2COO]-and[R 1COO]-can be used to determine the type of fatty acid,and[M-CH 3-R 2CO]-ion can be used to identify the linked site of fatty acid.Finally,12 phosphatidylcholines were detected.