凝胶排阻色谱法研究阿胶中蛋白质及多肽相对分子质量分布规律___

目的：建立阿胶中水溶性成分凝胶排阻色谱分析方法，研究阿胶中蛋白质及多肽的相对分子质量分布规律，为阿胶质量评价提供依据。方法：采用TSK-GEL G4000PWXL(7.8 mm×300 mm，平均孔径500 A°),流动相为0.05 mol·L-1磷酸盐缓冲液(pH=7.2),流速0.4 mL·min-1，检测波长280 nm，以β-淀粉酶、胰岛素为标记物，室温下测定16个企业生产的40批阿胶样品，并应用TQ Analyst软件进行样品图谱主成分分析。结果：发现阿胶含有的蛋白质及多肽的相对分子质量主要集中在6×103～2×105，不同企业的产品中蛋白质及多肽的相对分子质量分布有一定规律。结论：该方法准确、可靠，具有良好的重复性和稳定性，可用于阿胶中蛋白质及多肽的分析。测定结果反映出不同企业间，甚至同一企业不同批次产品有差异。提示阿胶原料与生产工艺值得注意，为保证阿胶质量稳定，应该进行生产过程控制。

AbstractObjective:To develop a gel exclusion chromatography method for study of relative molecular mass distribution of protein and peptide in donkey-hide glue.Methods:GPC was performed on a TSK-GEL G4000PWXL column (7.8 mm×300 mm, average pore size 500 A°) with 0.05 mol·L-1 phosphate buffered solution(pH 7.2) at the flow rate of 0.4 mL·min-1, and detection at 280 nm.Results:Relative molecular mass of protein and peptide in donkey-hide glue is in the ranges of 6× 103-2×105, and there are regular pattern in the relative molecular mass distribution of protein and peptide in donkey-hide glue.Conclusion:The developed method is accurate with high repeatability and stability, which can be used for the analysis of protein and peptide in donkey-hide glue. The result shows that there are difference in products among different manufacture, even among different batchs, which caused by different production technique.The control of product production process is very important to ensure the quality of donkey-hide glue.

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