牛Ⅰ型胶原的性能表征及标准样品研制

Characterization and development of reference material of bovine typeⅠcollagen

以牛Ⅰ型胶原作为研究对象,胶原标准样品研制为目标,对其红外光谱特征、三螺旋结构、热稳定性、氨基酸覆盖率、端肽残留、分子质量等主要性能指标进行分析,将羟脯氨酸含量作为判定依据对产品的均匀性进行分析;利用SDS-PAGE法对其稳定性进行分析;以8家实验室测定的羟脯氨酸含量进行定值和不确定度分析。结果表明,该产品的各项性能指标符合胶原蛋白的质量要求。该牛Ⅰ型胶原产品在95%置信区间内均匀性良好;在4℃放置2周、4周,25℃放置1周、2周,反复冻融1次、3次和5次及在-20℃放置6个月,产品稳定性良好;羟脯氨酸含量的定值结果为11.17%,置信度为95%时扩展不确定度为0.03(k=2)。因此,该牛Ⅰ型胶原可作为国家标准样品用于胶原产品的质量评价。

Bovine typeⅠcollagen was used as the research object.The main purpose was to develop the reference material of bovine typeⅠcollagen.The characterizations,such as infrared spectroscopy(IR),triple helix structure,thermal stability,amino acid sequence coverage,terminal peptide residue,molecular weight et al,were analyzed.The homogeneity and stability were evaluated using the content of hydroxyproline and the SDS-PAGE,respectively.The value assignment and uncertainty assessment were evaluated using the content of hydroxyproline from eight laboratories.The results indicated that bovine typeⅠcollagen met the requirement of collagen.The homogeneity of bovine typeⅠcollagen was excellent in confidence coefficient of 95%.And it had good stability within 2 and 4 weeks at 4℃,1 and 2 weeks at 25℃,repeated freezing and thawing 1,3 and 5 times,and 6 months at-20℃.The certified value of the hydroxyproline was 11.17%with the expanded uncertainty of 0.03 in confidence coefficient of 95%.Therefore,the bovine typeⅠcollagen could be used as national reference material for the quality evaluation of collagen products.