Quantitation of 1-stearoyl-2-arachidonoyl-sn-3-glycerol in human basophils via gas chromatography-negative ion chemical Ionization mass spectrometry
Quantitation of 1-stearoyl-2-arachidonoyl-sn-3-glycerol in human basophils via gas chromatography-negative ion chemical Ionization mass spectrometry
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DOI：
10.1006/abio.1996.0172
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发表时间：
1996-05-01
影响因子：
2.9
通讯作者：
MacGlashan, DW
中科院分区：
文献类型：
--
作者：
Hubbard, WC;Hundley, TR;MacGlashan, DW
由于缺乏具有足够灵敏度和选择性的测定方法，研究甘油二酯分子在人嗜碱性粒细胞信号转导中的作用受到阻碍。1-硬脂酰基-2-花生四烯酸酰基-sn-3-甘油转化为五氟苯甲酰基酯将电子捕获性质传递给二酰基甘油。甘油二酯的电子捕获衍生物适合气相色谱分析，并在负离子质谱条件下进行有限的碎片化，在m/z 838处产生强烈的分子阴离子。监测m/z 838以检测1-硬脂酰-2-花生四烯酸-sn-3-甘油，监测m/z 841以检测1-三氘硬脂酰-3-花生四烯酸-sn-2（用作内标物），为GC-MS定量测定人嗜碱性粒细胞中的内源性二酰甘油提供了分析基础。该测定在宽浓度范围内显示出优异的重现性，变化小于或等于10%。GC-MS分析具有高度选择性和极高灵敏度，每次进样内源性1-硬脂酰-2-花生四烯酸-sn-3-甘油的检测限小于或等于0.20 pg（类似于30 fmol）。从10（5）个刺激的人嗜碱性粒细胞中提取约400 fmol的二酰基甘油。(C)出版社：Academic Press，Inc.
Investigation of the role of diacylglycerol molecular species in signal transduction in human basophils has been impeded by the lack of an assay method with adequate sensitivity and selectivity. Conversion of 1-stearoyl-2-arachidonoyl-sn-3-glycerol to the pentafuorobenzoyl ester conveys electron-capture properties to the diacylglycerol. The electron-capture derivative of the diacylglycerol is amenable to gas chromatographic analysis and undergoes limited fragmentation under negative ion mass spectrometric conditions with generation of an intense molecular anion at m/z 838. Monitoring m/z 838 for detection of 1-stearoyl-2-arachidonoyl-sn-3-glycerol and m/z 841 for detection of 1-trideuterostearoyl-3-arachidonoyl-sn-2 employed as the internal standard provides the analytical basis for GC-MS quantitation of the endogenous diacylglycerol in human basophils. The assay displays excellent reproducibility over a wide range of concentrations with variations less than or equal to 10%. The GC-MS assay is highly selective and exquisitely sensitive with a detection limit of less than or equal to 0.20 pg (similar to 30 fmol) for endogenous 1-stearoyl-2-arachidonoyl-sn-3-glycerol per injection. Approximately 400 fmol of the diacylglycerol were extracted from 10(5) stimulated human basophils. (C) 1996 Academic Press, Inc.