Taguchi R, Hayakawa J, Takeuchi Y, Ishida M
Faculty of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya City, Aichi 467, Japan.
J Mass Spectrom. 2000 Aug;35(8):953-66. doi: 10.1002/1096-9888(200008)35:8<953::AID-JMS23>3.0.CO;2-4.
A phospholipid mixture extracted from cultured cells was directly analyzed by capillary (Cap) liquid chromatography (LC)/electrospray ionization (ESI) mass spectrometry (MS). Using a quadrupole mass spectrometer, we analyzed positive molecular ions, negative molecular ions, positive fragment ions and negative fragment ions under four different functions. In the analysis of the elution patterns of the phospholipids, a two-dimensional map, in which the first dimension is elution time and the second dimension is mass, proved useful. Consequently, four different maps can be obtained by each of four different functions. Among them, from negative fragment ions at high cone voltage in the negative ion mode, ions that originated from acyl fatty acid and phosphorylcholine, phosphorylethanolamine and cyclic inositol phosphate can be detected at specific elution times. The map from positive fragment ions at high cone voltage in the positive ion mode indicated ions such as diradylglycerol and derivatives of 1-alkyl or 1-alkenyl cyclic phosphatidic acid from phosphatidylethanolamine (PE), and phosphorylcholine from choline-containing phospholipids. The map produced from positive molecular ions indicated choline-containing phospholipids such as phosphatidylcholine, sphingomyelin, lysophosphatidylcholine and PE. The map of negative molecular ions effectively indicated acidic phospholipids such as phosphatidylinositol. We were able to obtain more than 500 molecular species of phospholipids by this method within a few hours immediately after extraction from culture cells using a mixture of chloroform and methanol (2:1). In this context, we concluded that the combination of Cap-LC and ESIMS seems to be very effective in the analysis of phospholipid classes and their molecular species.
从培养细胞中提取的磷脂混合物通过毛细管（Cap）液相色谱（LC）/电喷雾电离（ESI）质谱（MS）进行直接分析。使用四极杆质谱仪，我们在四种不同功能下分析了正分子离子、负分子离子、正碎片离子和负碎片离子。在磷脂洗脱模式的分析中，一个二维图谱被证明是有用的，其中第一维是洗脱时间，第二维是质量。因此，通过四种不同功能中的每一种都可以获得四种不同的图谱。其中，在负离子模式下高锥电压的负碎片离子中，可以在特定洗脱时间检测到源自酰基脂肪酸和磷酸胆碱、磷酸乙醇胺和环肌醇磷酸的离子。正离子模式下高锥电压的正碎片离子图谱显示了诸如二酰基甘油和来自磷脂酰乙醇胺（PE）的1-烷基或1-烯基环磷脂酸衍生物，以及来自含胆碱磷脂的磷酸胆碱等离子。正分子离子产生的图谱显示了含胆碱的磷脂，如磷脂酰胆碱、鞘磷脂、溶血磷脂酰胆碱和PE。负分子离子图谱有效地显示了酸性磷脂，如磷脂酰肌醇。使用氯仿和甲醇（2:1）的混合物从培养细胞中提取后，我们能够在几小时内通过这种方法获得500多种磷脂分子种类。在此背景下，我们得出结论，Cap-LC和ESI-MS的组合在磷脂类别及其分子种类的分析中似乎非常有效。
