Division of Bioorganic Chemistry and Molecular Pharmacology, Department of Internal Medicine, Washington University School of Medicine, St Louis MO 63110, USA.
J Lipid Res. 2010 Jun;51(6):1599-609. doi: 10.1194/jlr.D004077. Epub 2010 Feb 1.
Recently, we used the favorable properties of 9-aminoacridine (9-AA) as matrix for the quantitative analysis of acidic metabolites and glycerophospholipids from extracts of biological materials [Sun, G., Yang, K., Zhao, Z., Guan, S., Han, X., and Gross, R.W. (2007) A shotgun metabolomics approach for rapid analysis of negatively-charged water-soluble cellular metabolites from mouse heart tissue. Anal. Chem. 79: 6629-6640; Sun, G., Yang, K., Zhao, Z., Guan, S., Han, X., and Gross, R.W. (2008) Matrix-assisted laser desorption/ionization-time of flight mass spectrometric analysis of cellular glycerophospholipids enabled by multiplexed solvent dependent analyte-matrix interactions. Anal. Chem. 80: 7576-7585.] by MALDI-MS. Herein, we extend this discovery and identified the selective desorption/ionization of sulfatides over other examined anionic lipids present in lipid extracts of biological samples by MALDI-MS using 9-AA as matrix. Through this approach, a high throughput method for the quantitative analysis of low to very low abundance sulfatide molecular species directly from crude lipid extracts has been developed. This method possessed a linear dynamic range of over 1,000-fold, a detection limit at the high attomole level, and a reproducibility of approximately 10% deviation. Many potential factors that might affect the quantitation of sulfatide species employing the method were examined and their effects were found to be negligible within experimental error. Collectively, these results demonstrate a powerful high throughput method for the measurement of sulfatides directly from extracts of biological samples, facilitating the study of sulfatide metabolism, trafficking, and homeostasis in health and disease.
最近,我们利用 9-氨基吖啶(9-AA)的有利性质,作为从生物材料提取物中定量分析酸性代谢物和甘油磷脂的基质[Sun, G., Yang, K., Zhao, Z., Guan, S., Han, X., and Gross, R.W. (2007) 一种用于快速分析小鼠心脏组织中带负电荷水溶性细胞代谢物的 shotgun 代谢组学方法。分析化学 79: 6629-6640; Sun, G., Yang, K., Zhao, Z., Guan, S., Han, X., and Gross, R.W. (2008) 通过基质辅助激光解吸/电离-飞行时间质谱分析细胞甘油磷脂的基质辅助激光解吸/电离-飞行时间质谱分析,通过多重溶剂依赖的分析物-基质相互作用实现。分析化学 80: 7576-7585。] 通过 MALDI-MS。在此,我们扩展了这一发现,并通过 MALDI-MS 使用 9-AA 作为基质,鉴定了在生物样品脂质提取物中存在的其他阴离子脂质选择性的硫酸酯的解吸/离子化。通过这种方法,已经开发出一种从粗脂质提取物中直接定量分析低至非常低丰度硫酸酯分子的高通量方法。该方法具有超过 1000 倍的线性动态范围、高阿特莫尔水平的检测限和约 10%偏差的重现性。研究了许多可能影响采用该方法定量硫酸酯的潜在因素,发现其影响在实验误差范围内可忽略不计。总的来说,这些结果表明了一种强大的高通量方法,可直接从生物样品提取物中测量硫酸酯,促进了硫酸酯代谢、运输和健康与疾病中的动态平衡的研究。