凝胶过滤色谱法测定蛋白胨中多肽的相对分子质量分布

Determination of relative molecular mass distribution of polypeptides in peptone by gel filtration chromatography

目的建立凝胶过滤色谱法测定蛋白胨中多肽相对分子质量的方法。方法样品以水作为提取剂,磷酸二氢钠与磷酸氢二钠溶液作为流动相(pH6.8),流速0.8mL/min,用BioSeP5μmSEC-s2000柱进行分离,由GPC软件通过线性关系计算得出样品的具体相对分子质量。结果相对分子质量在451~150000之间时,相对分子质量的对数与保留时间呈曲线关系,相关系数为0.9971,方法的精密度、重复性、准确度、稳定性的相对标准偏差均在3%之内。结论该方法操作简单、灵敏度高,适用于多肽类物质相对分子质量分布区间的确定。

Objective To establish a method for determination relative molecular mass of polypeptides in peptone by gel filtration chromatography. Methods The sample was separated by BioSeP 5 μm SEC-S2000 column at a flow rate of 0.8 mL/min using water as the extraction agent, sodium dihydrogen phosphate and sodium disodium phosphate solution as the mobile phase(pH 6.8). The specific relative molecular weight of the sample was calculated by GPC software through linear relationship. Results When the relative molecular mass was between 451 and 150000, the logarithm of the relative molecular mass is curving with the retention time, and the correlation coefficient was 0.9971. The relative standard deviation of the precision, repeatability, accuracy and stability of the method were all within 3%. Conclusion The method is simple, sensitive and suitable for determining the distribution interval of relative molecular weight of peptides.