动物组织中羟脯氨酸测定方法的建立及初步应用

Assay of Hydroxyproline in Animal Tissues and Preliminary Application

目的：建立动物组织中的羟脯氨酸测定方法，以之估计胶原含量并衡量组织愈合程度。方法：研究显色温度、氧化时间、样品中盐浓度以及对 二甲氨基苯甲醛（ Ｐ Ｄ Ａ Ｂ） 浓度对羟脯氨酸测定的影响，据此建立动物组织中的羟脯氨酸测定方法，并初步应用于实验动物犬贲门胶原含量的测定。结果：根据优化的实验结果，羟脯氨酸测定的条件为室温氧化４ ｍｉｎ ，１０ ％ Ｐ Ｄ Ａ Ｂ，１００ ℃显色２ ｍｉｎ 。该法回收率为９８ ％ ，变异系数为４ ％ 。结论：该法重复性好，可信度高，可作为临床判定愈合程度的一个定量指标。

Objective: The experiment was designed to establish an assay system to determine the hydroxyproline; Hyp in animal tissues,through which the collagen contents and the healing level of repair tissues were estimated. Methods:The impact of colouration temperature, oxidation time ,salt concentration of the sample, and concentration of para-dimethyl-amino-benzalde-hyde (PDAB) on the assay of Hyp was studied and a method for detecting the Hyp in animal tissues was established. This method was used in determination of collagen contents in the cardia of experimental dogs. Results:The optimal condition for Hyp detection consisted of oxidation for 4?minutes at room temperature,10%PDAB,incubation at 100?℃ for 2?minutes for colouration. The recovery and CV of this method were 98%and 4.0%,respectively. Conclusion:This method may be used as a reliable quantitative method to judge the healing level of repair tissues.