Separation and purification of pig femoral collagen antihyper-tensive peptides by ion exchange chromatography
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摘要: 猪股骨酶解液经超滤、凝胶层析初步分离后,为了得到高活性且纯度较高的降血压肽,采用离子交换层析对凝胶层析分离后的高活性组分进一步分离。通过对洗脱液p H、离子强度、流速三个因素进行研究,确定了最佳分离条件:以p H=6.0、0.05mol/L磷酸盐缓冲液为A液、B液为含1mol/LNa Cl的p H=6.0、0.05mol/L磷酸盐缓冲液,洗脱流速为0.8m L/min。结果显示:离子交换层析分离得3个组分,其中组分1的活性最高,其IC50值为0.1012mg/m L;再利用凝胶层析将组分1进行脱盐,其峰2的IC50值达到0.0836mg/m L,脱盐率达到86.60%±0.5%。Abstract: In order to obtain high activity and purity of antihypertensive peptides, the method of ion exchange chromatography was used to separat the solution after the preliminary separation and charactenization by ultrafiltration and gel permeation chromatography. The eluent p H, ionic strength, velocity of the three factors were studied. The optimal conditions for analysis were as follows : elution of p H = 6. 0, the flow rate was0.8m L/min, the ionic strength was 1.0mol/L. The results showed that the fraction was then purified by ion exchange chromatography into 3 peaks, in which peak 1 had the strongest ACE inhibitory activity with an IC50 of 0.1012mg/m L. And the peak 1 was further desalinationted by gel permeation chromatography to obtain 4peaks, and the peak 2 had the highest ACE-inhibitory activity with an IC50 of 0.0836mg/m L, the desalination rate 86.60%±0.5%.
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