凝胶色谱测定核桃肽分子量方法比较

本文研究了核桃肽分子量的凝胶色谱色谱检测方法,对比两种测定肽类分子量大小常用的凝胶色谱柱:(1)国家标准GB/T22492-2008《大豆肽粉》以及GB/T 22729-2008《海洋鱼低聚肽粉》中推荐的TSK-GEL G2000 SWXL 7.8 mm×300 mm色谱柱;(2)商务部行业标准SB/T 10634-2011《淡水鱼胶原蛋白肽粉》中推荐的Superdex Peptide HR 10/30色谱柱。本文通过对四种单一标准品以及四种不同分子量大小的核桃肽样品在两种色谱条件中的结果对比分析,得出以下结论:TSK-GEL G2000 SWXL 7.8 mm×300 mm色谱柱推荐条件能在30 min内完成核桃肽的分离,检测线性良好,R2=0.97,色谱图峰形良好清晰,检测结果在10000 u以及1000 u区间上偏大;Superdex Peptide HR 10/30色谱柱推荐条件在80 min内完成核桃分离,检测线性较佳,R2=0.99,色谱图峰形出现裂峰与宽峰,检测结果在10000 u以及1000 u区间上偏小。

Molecular Comparison of Gel-permeation Chromatography Methods to Determine the Weight of Walnut Peptides

Methods involving gel-permeation chromatography used to determine the molecular weight of walnut peptides were studied by comparing two commonly used gel columns. These included a TSK-GEL G2000 SWXL 7.8 mm × 300 mm column, recommended by the national standard GB/T 22492-2008 Soy Peptide Powder and GB/T 22729-2008 Oligopeptide Powder of Marine Fish, and a Superdex Peptide HR 10/30 column, recommended by the industry standard SB/T 10634-2011 Freshwater Fish Collagen Peptide Powder by the Ministry of Commerce. Data were measured, analyzed, and compared for four pure standards and four walnut peptide samples with different molecular weights in each of the two chromatographic conditions. The results showed that for TSK- GEL G2000 SWXL 7.8 mm × 300 mm column, the separation of walnut peptides was achieved within 30 min under the recommended conditions. The results also showed a good linear relationship, with a correlation coefficient of 0.97 and a clear peak shape. The measured molecular weights were higher than the actual values in the regions of >10,000 u and <1,000 u. For the Superdex Peptide HR 10/30 column in comparison, the separation was achieved after 80 min, however the results showed a better linear relationship, with a correlation coefficient of 0.99; however, spilt peaks and broad peaks appeared in the chromatogram. Moreover, in this system, the measured molecular weights were lower than the actual values in the regions of >10,000 u and <1,000 u.