天狼星红染色法体外定量测定水生动物源胶原的初步研究

A Preliminary Study on the Quantitative Determination of Collagen of Aquatic Animal Origin by Using Sirius Red Staining in Vitro

Collagen Assay是基于天狼星红与胶原特异性结合的原理而设计的一款新型天然胶原定量测定试剂盒。该产品已证实对于哺乳动物来源的可溶性胶原具有较好的测定效果,且方法简便。本研究尝试利用该试剂盒开展鱼类和两栖动物类胶原的定量测定,评价其测定效果及相关影响因素。测定精密度和准确度分析结果表明,该试剂盒具有较好的测定准确性、数据重现性和较强的抗杂蛋白干扰性。在此基础上,进一步开展了胶原热变性和氨基酸组成对测定结果的影响。结果表明,该试剂盒测定结果受胶原热变性程度的影响。随着胶原热变性程度的增加,其测定值逐渐减小且二者之间存在显著的线性相关性。该研究结果对于Sircol试剂盒在非哺乳动物胶原定量测定中的应用具有指导价值。

Based on the principle of the combination of Sirius red and collagen,Collagen Assay reagent kit is specially designed as a new reagent kit for the quantitative determination of the soluble collagen from mammalian.In this study,the reagent kit was used for the quantitative determination of collagen in fish and amphibian,of which the measurement results and influencing factors were studied.The reagent kit's good measurement accuracy,data reproducibility and strong anti protein interference feature have been showed in this precision and accuracy analysis.Base on the analysis and studies,a further study was carried out about the effect of thermal denaturation and amino acid composition of collagen on the determination results.The analysis showed that the measure result of the kit was influenced by the degree of thermal denaturation of collagen;The greater the degree of thermal denaturation of collagen,the smaller the measurement value of the kit,and there was a significant linear correlation between the two.This study has a directive value for the application of Sircol kit in the quantitative determination of nonmammalian collagen.