Quantitative determination of cyclic phosphatidic acid and its carba analog in mouse organs and plasma using LC-MS/MS
Quantitative determination of cyclic phosphatidic acid and its carba analog in mouse organs and plasma using LC-MS/MS
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DOI：
10.1016/j.jchromb.2018.01.002
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发表时间：
2018-02-15
影响因子：
3
通讯作者：
Murakami-Murofushi, Kimiko
中科院分区：
文献类型：
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作者：
Shimizu, Yoshibumi;Ishikawa, Masaki;Murakami-Murofushi, Kimiko
环磷脂酸 (cPA) 是溶血磷脂酸的类似物，参与许多细胞过程的调节。定量 cPA 分子种类的灵敏且特异的方法对于研究 cPA 的生理和病理生理作用非常重要。在此，我们开发了一种基于液相色谱-串联质谱 (LC-MS/MS) 的定量方法，用于同时检测具有各种脂肪酸（16:0、18:0、18:1 和 18:2）的 cPA 物种以及 2-carba-cPA（一种化学合成的 cPA 类似物）。使用反相C18柱进行色谱分析。使用三重四极杆质谱仪检测 cPA 物质。使用 cPA 17:0 作为内标。日内和日间精度值 (CV%) 均在 10% 以内。每种cPA 的检测线性范围为0.01 μg/mL 至5 μg/mL，相关系数为0.998 或更高。所开发的方法应用于小鼠血浆和器官中 cPA 物种的定量。研究表明，与对照小鼠相比，经 cuprizone 治疗的多发性硬化症模型小鼠大脑中 cPA 16:0、18:0 和 18:1 的浓度显着降低。这些发现对于理解 cPA 在与多发性硬化症相关的神经退行性过程中的作用可能很重要。
Cyclic phosphatidic acid (cPA), an analog of lysophosphatidic acid, is involved in the regulation of many cellular processes. A sensitive and specific method to quantify the molecular species of cPA is important for studying the physiological and pathophysiological roles of cPA. Here, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based quantification method for the simultaneous detection of cPA species having various fatty acids (16:0, 18:0, 18:1, and 18:2) as well as 2-carba-cPA, a chemically synthesized analog of cPA. Chromatography was performed using a reversed-phase C18 column. cPA species were detected using a triple quadrupole mass spectrometer. cPA 17:0 was used as an internal standard. Intra- and interday precision values (CV%) were within 10%. The linear range of detection for each cPA species was 0.01 mu g/mL to 5 mu g/mL, with correlation coefficients of 0.998 or higher. The developed method was applied to the quantification of cPA species in mouse plasma and organs. The concentrations of cPA 16:0, 18:0, and 18:1 were revealed to be significantly reduced in the brains of cuprizone-treated mice, a model of multiple sclerosis, compared with control mice. These findings could be important for understanding the roles of cPA in the neurodegenerative processes associated with multiple sclerosis.