乙醇系统HPLC分析PTC－氨基酸的条件优化

Optimization of Phenylthiocarbamyl (PTC) Amino Acid Serarations in Reversed Phase HPLC Based on Ethanol

首次报道用乙醇系统分析ＰＴＣ－氨基酸的新方法中各衍生物获得最佳分离的建立过程。ＰＴＣ－氨基酸衍生后溶于Ａ溶液，然后进样于４μｍＮｏｖａＰａｋＣ１８柱（３．９ｍｍ×１５０ｍｍ）。系统的优化步序包括全面调控流动相的ｐＨ值与ＴＥＡ浓度、乙醇梯度程序、柱温等诸多影响ＨＰＬＣ色谱行为的因素。最适条件为：Ａ溶液含０．１４ＭＧ酸钠、０．７５ｍｌ／ＬＴＥＡ、ＰＨ６．３５；Ｂ溶液为１００％乙醇；柱温３０℃。通过优化的乙醇梯度最终在约４４ｍｉｎ内将１５种ＰＴＣ－氨基酸很好地分离。此法可用于替代代表新科技水平的ＰＴＣ－氨基酸乙腈分析系统。

An HPLC system has been developed for separation of phenylthiocarbamyl (PTC)amino acids based on utilizing ethanol as organic eluent. The derivatized PTC amino acids are dissolved in solvent A for injection onto the 4 μm Nova Pak C18 column(3.9mm×150mm) . Optimization of the prodecure involves systematically controlling comprehensive factors influencing HPLC performance including PH, triethylamine (TEA) concentration in the mobile phase, program gradient of ethanol and column temperature.Optimized solvent A contains 0.14 M sodium acetate plus 0.75 ml/liter TEA adjusted to PH 6.35 with acetate, and solvent B consists of 100% ethanol. Fine resolution of 15 PTC amino acids is achieved at 30℃ in about 44 min utilizing gradient of increasing concentration of solvent B in solvent A.This approach to amino acid analysis offers select advantages with respect to the state-of-the-art PTC amino acid analysis based on acetonitrile.