Ito A, Uoji H, Mori Y
Anal Biochem. 1985 Dec;151(2):510-4. doi: 10.1016/0003-2697(85)90212-x.
An enzymatic method using 4-hydroxyproline 2-epimerase (EC 5.1.1.9), D-amino acid oxidase (EC 1.4.3.3.), and a colorimetric reagent of the mixture of Ti(IV) and 4-(2-pyridylazo)-resorcinol was found to be useful in the spectrophotometric assay for hydroxyproline in collagenous tissue hydrolysates. It was possible to determine free hydroxyproline in 250 microliter of tissue hydrolysates at concentrations ranging from 1 to 12.5 micrograms. The coefficient of variation was less than 3%, and the correlation between the proposed method and the colorimetric method (K.I. Kivirikko, O. Laitinen, and D. J. Prockop (1967) Anal. Biochem. 19, 249-255) was 0.973. Thus, this method is convenient because it requires less time than the above colorimetric method, and is also reproducible and sensitive.
已发现一种酶法，该方法使用4-羟基脯氨酸2-表异构酶（EC 5.1.1.9）、D-氨基酸氧化酶（EC 1.4.3.3.）以及钛（IV）和4-（2-吡啶偶氮）间苯二酚混合物的比色试剂，可用于分光光度法测定胶原组织水解物中的羟脯氨酸。能够在250微升组织水解物中测定浓度范围为1至12.5微克的游离羟脯氨酸。变异系数小于3%，且所提出的方法与比色法（K.I. 基维里科、O. 莱蒂宁和D.J. 普罗科普（1967年）《分析生物化学》19卷，249 - 255页）之间的相关性为0.973。因此，该方法很方便，因为它比上述比色法所需时间更少，并且具有可重复性和灵敏性。
