Tan Bo, Yu Y William, Monn M Francesca, Hughes H Velocity, O'Dell David K, Walker J Michael
The Gill Center for Biomolecular Science, Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN 47405, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Sep 15;877(26):2890-4. doi: 10.1016/j.jchromb.2009.01.002. Epub 2009 Jan 9.
Great effort has been devoted to characterize signaling lipids in central nervous system. This has led to a search for novel strategies to characterize hitherto unknown lipid compositions. Here we developed two methods, one for identification and one for quantification, for N-acyl amino acids, a novel lipid family. The identification method contains a series of purification steps followed by nano-LC/MS/MS and high-throughput screening of the datasets with a potent search algorithm based on fragment ion analysis. MS/MS spectra with good quality can be obtained with 150 fmol of targeted lipids on column with our nano-LC/MS/MS. More than one thousand mass spectra generated using the information dependent acquisition mode of Analyst QS software can be analyzed in 1 min using our home built software. The quantification method utilized the multiple reaction monitoring mode in Analyst software to measure the endogenous levels of N-acyl amino acids in rat brain. Using these two methods we were able to identify and quantify 11 previously reported N-acyl amino acids with endogenous levels ranging from 0.26 to 333 pmol g(-1) wet rat brain.
人们付出了巨大努力来表征中枢神经系统中的信号脂质。这促使人们寻找新的策略来表征迄今未知的脂质组成。在此，我们针对一种新型脂质家族——N-酰基氨基酸，开发了两种方法，一种用于鉴定，一种用于定量。鉴定方法包含一系列纯化步骤，随后进行纳升液相色谱/串联质谱分析，并使用基于碎片离子分析的强大搜索算法对数据集进行高通量筛选。使用我们的纳升液相色谱/串联质谱，在柱上有150飞摩尔的目标脂质时，就能获得高质量的串联质谱图。使用我们自行开发的软件，1分钟内就能分析使用Analyst QS软件的信息依赖采集模式生成的一千多个质谱图。定量方法利用Analyst软件中的多反应监测模式来测量大鼠脑中N-酰基氨基酸的内源性水平。使用这两种方法，我们能够鉴定并定量11种先前报道的N-酰基氨基酸，其在湿重大鼠脑中的内源性水平范围为0.26至333皮摩尔/克。
