Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Anal Chem. 2010 Nov 1;82(21):8794-9. doi: 10.1021/ac1021826. Epub 2010 Oct 14.
Cardiolipin (CL) and bis(monoacylglycero)phosphate (BMP) are unique lipid structures with important biological roles for mitochondrial integrity and endolysosomal degradation, respectively. They are synthesized from common precursors, phosphatidylglycerol (PG) and phosphatidic acid (PA). Here we present a rapid method for the simultaneous quantification of BMP, CL, PG, and PA using hydrophilic interaction chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-MS/MS). HILIC provides coelution of lipid species and their internal standards required for accurate quantification. Coelution leads to isotope overlap of lipid species which was successfully corrected. This assay was validated in mouse heart tissue and primary human skin fibroblasts. It shows reproducibility and limits of detection sufficient for biomarker studies contributing to basic research on BMP and CL metabolism.
心磷脂 (CL) 和双 (单酰基甘油) 磷酸酯 (BMP) 是具有重要生物学功能的独特脂质结构，分别在心粒体完整性和内溶酶体降解中发挥作用。它们分别由共同的前体磷脂酰甘油 (PG) 和磷脂酸 (PA) 合成。本文介绍了一种使用亲水相互作用色谱-电喷雾串联质谱 (HILIC-MS/MS) 同时定量分析 BMP、CL、PG 和 PA 的快速方法。HILIC 可使脂质种类及其内标共洗脱，从而实现准确的定量分析。共洗脱导致脂质种类的同位素重叠，我们成功地对此进行了校正。该方法在小鼠心脏组织和原代人皮肤成纤维细胞中进行了验证，具有良好的重现性和足够的检测限，可用于 BMP 和 CL 代谢的生物标志物研究，为基础研究做出贡献。
