Direct quantitative analysis of lysophosphatidic acid molecular species by stable isotope dilution electrospray ionization liquid chromatography-mass spectrometry
Direct quantitative analysis of lysophosphatidic acid molecular species by stable isotope dilution electrospray ionization liquid chromatography-mass spectrometry
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DOI：
10.1006/abio.2001.5063
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发表时间：
2001-05-15
影响因子：
2.9
通讯作者：
Tigyi, GJ
中科院分区：
文献类型：
--
作者：
Baker, DL;Desiderio, DM;Tigyi, GJ
为了更好地了解溶血磷脂酸（LPA）在生理和病理生理中的作用，有必要准确测定生物样品中LPA的分子种类和数量。我们已经开发了一种稳定同位素稀释、液相色谱-质谱法用于直接定量分析1-酰基-LPA。该方法利用氘标记的内标物LPA（18：0-d（35））和使用酸性丁醇的单次液-液萃取（其允许>95%的LPA回收率），随后进行在线正相液相色谱-质谱法。该方案允许对生物样品中存在的单个1-酰基-LPA物质进行准确、灵敏和可重现的分析。通过分析人类志愿者血浆和血清中的LPA物质证明了该测定法的实用性。EDTA血浆中的总LPA在雄性中为0.61 +/- 0.14 μ M，在雌性中为0.74 +/- 0.17 μ M，在25 ℃孵育24小时后增加至0.91 +/- 0.23和0.99 +/- 0.38 μ M，血清中的总LPA在雄性中为0.85 +/- 0.22 μ M，在雌性中为1.57 +/- 0.56 μ M，在25 ℃孵育24小时后增加至4.78 +/- 0.89和5.57 +/- 0.73 μ M。(C)北京：科学出版社.
In order to better understand the role of lysophosphatidic acid (LPA) in physiology and pathophysiology, it is necessary to accurately determine the molecular species and amounts of LPA in biological samples. We have developed a stable-isotope dilution, liquid chromatography-mass spectrometry assay for the direct quantitative analysis of 1-acyl-LPA, This method utilizes a deuterium-labeled internal standard, LPA (18:0-d(35)), and a single liquid-liquid extraction with acidic butanol that allows >95% recovery of LPA, followed by online normal-phase liquid chromatography-mass spectrometry. This protocol allows for the accurate, sensitive, and reproducible analysis of the individual 1-acyl-LPA species present in biological samples. The utility of the assay is demonstrated through the analysis of LPA species in plasma and serum from human volunteers. Total LPA in EDTA plasma was 0.61 +/- 0.14 muM in males and 0.74 +/- 0.17 muM in females, which increased to 0.91 +/- 0.23 and 0.99 +/- 0.38 muM after incubation for 24 h at 25 degreesC, Total LPA in serum was 0.85 +/- 0.22 muM in males and 1.57 +/- 0.56 muM in females, which increased to 4.78 +/- 0.89 and 5.57 +/- 0.73 muM after incubation for 24 h at 25 degreesC. (C) 2001 Academic Press.