Bhown A S, Bennett J C
Anal Biochem. 1985 Nov 1;150(2):457-62. doi: 10.1016/0003-2697(85)90535-4.
An improved and very simple procedure for thiazolinone conversion to thiohydantoin derivatives and their separation by reverse-phase high-pressure liquid chromatography is described. Trifluoroacetic acid (10%) in ethyl acetate has been employed as a conversion reagent to circumvent the deamidation of acid amides and methylation of acidic amino acids, with a concomitant increase in the detection limits of these residues. Additionally, a very simple procedure has been developed for the separation of phenylthiohydantoin (PTH) derivatives of amino acids. The system takes advantage of the computer-controlled precise mixing of the solvents A and B to achieve accurate pH and thus avoid the necessity of pH adjustment of a buffer. The procedure is simple and highly reproducible, and separates all the 20 known PTH amino acids. The efficiency of the method has been examined on synthetic and natural proteins/peptides, in manual and autoconversion systems, over a period of more than 18 months.
本文描述了一种改进的、非常简单的将噻唑啉酮转化为硫代乙内酰脲衍生物及其通过反相高压液相色谱分离的方法。已使用乙酸乙酯中的三氟乙酸（10%）作为转化试剂，以避免酸酰胺的脱酰胺作用和酸性氨基酸的甲基化，同时提高这些残基的检测限。此外，还开发了一种非常简单的方法用于分离氨基酸的苯硫代乙内酰脲（PTH）衍生物。该系统利用计算机控制的溶剂A和B的精确混合来实现精确的pH值，从而避免了缓冲液pH调节的必要性。该方法简单且重现性高，可分离所有20种已知的PTH氨基酸。在超过18个月的时间里，已在合成和天然蛋白质/肽、手动和自动转化系统中检验了该方法的效率。
