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* Department of Applied Biological Science, Fukuyama University, Higashimura, Fukuyama, 729-0292, Japan
Department of Functional Molecular Science, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8551, Japan
Faculty of Pharmaceutical Sciences, University of Tokushima, 1-78-1 Shomachi, Tokushima, 770-8505, Japan
1 To whom correspondence should be addressed. e-mail: tamot{at}fubac.fukuyama-u.ac.jp
Lysophosphatidic acid (LPA) is a lipid mediator that may play an important role in wound healing, embryonic development, and progression of cancer. Here, we report a procedure for the quantification of LPA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The method is based on a characteristic mass shift with total charge change (from 2 to +1) of the phosphate species due to 1:1 complexation of LPA2 with a dinuclear zinc (II) complex {1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato dizinc(II) complex; Zn2L3+} at physiological pH. The monocationic complex [LPA2-Zn2L3+]+ was detected in the positive mode, in which no other signal of cation adducts of LPA2 was observed. The detection limit of 18:1 LPA by this method was 0.1 pmol on a sample plate. The intensity ratio of [LPA2-Zn2L3+]+ against an internal standard [17:0 LPA2-Zn2L3+]+ increased linearly with their molar ratio. Based on the relative intensities of complex ions, we determined the amounts of LPA homologs in an egg white by this method; the results obtained were in good agreement with those by gas liquid chromatography.
This sensitive and convenient procedure for LPA-specific detection is useful for the quantification of LPA homologs occurring in biological materials.
Abbreviations: DHB, 3,5-dihydroxybenzoic acid; ESI, electrospray ionization; LPA, lysophosphatidic acid; MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; PC, phosphatidylcholine; THAP, 2,4,6-trihydroxy-acetophenone; Zn2L3+, 1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato dizinc(II) complex
Supplementary key words egg white matrix-assisted laser desorption/ionization time-of-flight mass spectrometry serum
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