Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2 Canada.
Anal Chem. 2010 Nov 1;82(21):8789-93. doi: 10.1021/ac102146g. Epub 2010 Oct 14.
We have developed a new isotope labeling method, based on the use of isotope-coded p-dimethylaminophenacyl (DmPA) bromide as a reagent, combined with liquid chromatography-mass spectrometry (LC-MS) for high-performance metabolome analysis with a focus on profiling carboxylic acid-containing metabolites. Derivatization is simple, fast (1 h plus 30 min for quenching the reaction), and applicable to a wide range of carboxylic acids with a high yield and little or no side reaction products. This labeling method is demonstrated to be not only effective in introducing an isotope tag for accurate metabolite quantification but also improving the chromatographic retention of the metabolites in reversed-phase (RP) LC, enhancing ESI efficiency by 2-4 orders of magnitude, and facilitating the identification of metabolite peaks in LC-MS. In triplicate experiments of a 1:1 ratio of (13)C-/(12)C-DmPA labeled human urine, we were able to detect 2671, 2546, and 2820 ion pairs from metabolites containing one or more carboxylic acid groups.
我们开发了一种新的同位素标记方法，基于使用同位素编码的对二甲氨基苯乙酰基（DmPA）溴化物作为试剂，结合液相色谱-质谱（LC-MS）进行高效代谢组学分析，重点是对含羧酸代谢物进行分析。衍生化简单、快速（反应淬灭需要 1 小时加 30 分钟），适用于广泛的羧酸，具有高收率和很少或没有副反应产物。该标记方法不仅有效用于引入同位素标签进行准确的代谢物定量，而且改善了反相（RP）LC 中代谢物的色谱保留，通过 2-4 个数量级提高电喷雾效率，并有助于在 LC-MS 中鉴定代谢物峰。在（13）C/（12）C-DmPA 标记的人尿 1:1 比例的重复实验中，我们能够从含有一个或多个羧酸基团的代谢物中检测到 2671、2546 和 2820 个离子对。
