Quantitation of 5-lipoxygenase products by electrospray mass spectrometry: effect of ethanol on zymosan-stimulated production of 5-lipoxygenase products by human neutrophils.
Quantitation of 5-lipoxygenase products by electrospray mass spectrometry: effect of ethanol on zymosan-stimulated production of 5-lipoxygenase products by human neutrophils.
复制标题
通过电喷雾质谱法定量 5-脂氧合酶产物：乙醇对酵母聚糖刺激的人中性粒细胞产生 5-脂氧合酶产物的影响。
DOI：
10.1006/abio.1996.9885
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Murphy,RC
中科院分区：
文献类型：
--
作者：
Wheelan,P;Murphy,RC
开发了一种反相 HPLC/串联质谱方法 (LC/MS/MS)，用于定量白三烯 B4 (LTB4)、5-脂氧合酶产物、5-羟基二十碳四烯酸 (5-HETE) 以及 LTB4、20-羟基-LTB4 和 20-羧基-LTB4 的 ω-氧化代谢物。电喷雾产生的羧酸根阴离子被碰撞激活并分解为特异且丰富的产物离子，并使用多反应监测分析LTB4(m/z335 → 195)、20-羟基-LTB4(m/z351 → 195)、20-羧基-LTB4(m/z365 → 195)和5-HETE (m/z319 → 115）。在调理性酵母聚糖刺激人嗜中性粒细胞期间产生的LTB4的五氟苄酯/三甲基甲硅烷基醚衍生物的LC/MS/MS定量结果与气相色谱/MS定量结果的比较中，观察到线性相关性。所有代谢物的检测限均处于低皮克水平。该方法适用于在乙醇存在下对由酵母聚糖刺激的人中性粒细胞产生的 5-脂氧合酶产物进行定量。在生理相关浓度的乙醇下，受刺激的中性粒细胞产生的所有 5-脂氧合酶产物的产生均显着减弱。
A reverse-phase–HPLC/tandem mass spectrometric method (LC/MS/MS) was developed for the quantitation of leukotriene B4(LTB4), the 5-lipoxygenase product, 5-hydroxyeicosatetraenoic acid (5-HETE), as well as the ω-oxidation metabolites of LTB4, 20-hydroxy-LTB4, and 20-carboxy-LTB4. Electrospray-generated carboxylate anions were collisionally activated and decomposed to specific and abundant product ions and multiple reaction monitoring was used to analyze LTB4(m/z335 → 195), 20-hydroxy-LTB4(m/z351 → 195), 20-carboxy-LTB4(m/z365 → 195), and 5-HETE (m/z319 → 115). A linear correlation was observed in comparison of results obtained from quantitation by LC/MS/MS with quantitation by gas chromatography/MS of the pentafluorobenzyl ester/trimethylsilyl ether derivative of LTB4produced during opsonized zymosan stimulation of human neutrophils. Detection limits at the low picogram level were obtained for all metabolites. This method was applied to the quantitation of 5-lipoxygenase products produced from zymosan-stimulated human neutrophils in the presence of ethanol. At physiologically relevant concentrations of ethanol, production of all 5-lipoxygenase products generated by stimulated neutrophils was markedly attenuated.