Negro A, Garbisa S, Gotte L, Spina M
Anal Biochem. 1987 Jan;160(1):39-46. doi: 10.1016/0003-2697(87)90611-7.
A rapid and accurate reverse-phase high-performance liquid chromatography procedure for amino acid analysis of connective tissue proteins has been optimized. The method is based on quantitative dansyl chloride precolumn derivatization of protein hydrolyzates and chromatographic separation of the dansyl derivatives on Ultrasphere ODS C18 column. High molar uv absorption of Pro and Hyp derivatives, quantitation of low Des and Ide amounts (60 pmol), and good separation of all the amino acid derivatives (CV mostly less than 0.1%) including Hyl overcome the difficulties of other methods in producing reliable single run amino acid analysis of collagen and elastin. The use of fluorescence detector and the possibility of concentrating derivatized samples would give even higher sensitivity to the system. The procedure appeared to be suitable for single run analysis of other proteins, particularly those having an unbalanced amino acid composition.
一种用于结缔组织蛋白氨基酸分析的快速准确的反相高效液相色谱法已得到优化。该方法基于蛋白质水解产物的丹磺酰氯柱前定量衍生化以及丹磺酰衍生物在Ultrasphere ODS C18柱上的色谱分离。脯氨酸和羟脯氨酸衍生物的高摩尔紫外吸收、低量去甲氨基酸和异氨基酸（60皮摩尔）的定量以及包括羟赖氨酸在内的所有氨基酸衍生物的良好分离（变异系数大多小于0.1%）克服了其他方法在对胶原蛋白和弹性蛋白进行可靠的单次氨基酸分析时遇到的困难。荧光检测器的使用以及衍生化样品浓缩的可能性将使该系统具有更高的灵敏度。该方法似乎适用于其他蛋白质的单次分析，尤其是那些氨基酸组成不均衡的蛋白质。
