Department of Pharmaceutical Science, Alma Mater Studiorum, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy.
J Pharm Biomed Anal. 2010 Oct 10;53(2):207-11. doi: 10.1016/j.jpba.2009.12.031. Epub 2010 Jan 11.
The use of 2,5-dimethyl-1H-pyrrole-3,4-dicarbaldehyde as a precolumn derivatization reagent for HPLC analysis of amino acids is proposed. The compound reacts under mild conditions (10min at ambient temperature) with primary amino groups. The derivatization conditions to obtain quantitative reaction were optimised by considering different parameters (temperature, pH and reagent concentration) using l-Val as the model compound. The synthesized l-Val derivative was characterized by (1)H NMR and UV. The derivatives of 19 amino acids were separated by reversed-phase HPLC and detected at lambda=320nm. The method was applied successfully to the qualitative and quantitative analysis of commercial polyamino acid preparations.
提议使用 2,5-二甲基-1H-吡咯-3,4-二醛作为 HPLC 分析氨基酸的柱前衍生试剂。该化合物在温和条件下(在环境温度下反应 10 分钟)与伯氨基反应。通过考虑不同参数(温度、pH 值和试剂浓度),使用 l-Val 作为模型化合物,优化了获得定量反应的衍生化条件。合成的 l-Val 衍生物通过(1)H NMR 和 UV 进行表征。19 种氨基酸的衍生物通过反相 HPLC 分离,并在 lambda=320nm 处检测。该方法成功应用于商业多氨基酸制剂的定性和定量分析。