Charting molecular composition of phosphatidylcholines by fatty acid scanning and ion trap MS3 fragmentation
Charting molecular composition of phosphatidylcholines by fatty acid scanning and ion trap MS3 fragmentation
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DOI：
10.1194/jlr.d300020-jlr200
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发表时间：
2003-11-01
影响因子：
6.5
通讯作者：
Shevchenko, A
中科院分区：
文献类型：
--
作者：
Ekroos, K;Ejsing, CS;Shevchenko, A
采用混合四极杆飞行时间质谱联用多前体离子扫描和离子阱质谱联用MS3碎片化研究了磷脂酰胆碱（PCs）在总脂提取物中的分子组成。平行获取脂肪酸中50个酰基阴离子片段的前体离子谱（脂肪酸扫描），增加了PC检测的特异性和动态范围，并鉴定了单个PC物种的脂肪酸部分。随后在离子阱质谱仪上通过MS3碎片对检测到的PC峰进行分析，量化了它们的位置异构体的相对数量，从而在低皮摩尔水平上提供了PC池分子组成的最详细和全面的表征。与传统方法相比，该方法大大简化，并且不需要脂类或单个分子种类的初步分离，酶消化或化学衍生化。通过对人红细胞中pc分子组成的对比分析，验证了该方法的有效性。在Madin-Darby犬肾II细胞的总脂质提取物中，我们检测到46种具有独特脂肪酸组成的PC，并证明位置异构体的存在几乎使单个分子种的总数增加了一倍。-Ekroos， K. S. Ejsing， U. Bahr， M. Karas， K. Simons， A. Shevchenko。利用脂肪酸扫描和离子阱MS3破碎绘制磷脂酰胆碱分子组成图。
The molecular composition of phosphatidylcholines (PCs) in total lipid extracts was characterized by a combination of multiple precursor ion scanning on a hybrid quadrupole time-of-flight mass spectrometer and MS3 fragmentation on an ion trap mass spectrometer. Precursor ion spectra for 50 acyl anion fragments of fatty acids (fatty acid scanning) acquired in parallel increased the specificity and the dynamic range of the detection of PCs and identified the fatty acid moieties in individual PC species. Subsequent analysis of detected PC peaks by MS3 fragmentation on an ion trap mass spectrometer quantified the relative amount of their positional isomers, thus providing the most detailed and comprehensive characterization of the molecular composition of the pool of PCs at the low-picomole level. The method is vastly simplified, compared with conventional approaches, and does not require preliminary separation of lipid classes or of individual molecular species, enzymatic digestion, or chemical derivatization. The approach was validated by the comparative analysis of the molecular composition of PCs from human red blood cells.jlr In the total lipid extract of Madin-Darby canine kidney II cells, we detected 46 PC species with unique fatty acid composition and demonstrated that the presence of positional isomers almost doubled the total number of individual molecular species.-Ekroos, K., C. S. Ejsing, U. Bahr, M. Karas, K. Simons, and A. Shevchenko. Charting molecular composition of phosphatidylcholines by fatty acid scanning and ion trap MS3 fragmentation.