乙醇系统HPLC分析PTC－氨基酸的条件优化

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。

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

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.