肝水解肽的制备及相对分子质量分布研究

Study on preparation method and molecular weight distribution of polypeptide in heparolysate

目的研究肝水解肽的制备方法及相对分子质量(Mr)分布。方法采用碱化-酶解法制备肝水解肽,采用高效液相色谱法测定肝水解肽的Mr。结果采用碱化-酶解法制备肝水解肽收率稳定,优化的酶解法工艺参数为胰酶用量为0.5%、温度为55℃、pH为7.5、水解时间为6 h,在此条件下酶解多肽提取率平均为4.82%;肝水解肽Mr400～10 000的组分占73%左右。结论碱化-酶解法制备肝水解肽工艺简便可靠,能保持大部分多肽结构。

Purpose To study the preparation technique and to establish a method for determining the molecular weight （Mr） distribution of polypeptide in heparolysate. Methods The heparolysate was pre- pared by basification-enzymolysis method,and the Mr distribution was determined by HPLC. Results The optimum preparation data were as follows,the dosage of pancreatin 0.5% ,55 ℃ ,pH 7.5 and the time of enzymolysis 6 h. The average yield of heparolysate was 4.82% , in the Mr range of 400-10 000 D. There was iiaean relationship between the logarithm of Mr of peptide and the retention time （r = 0. 993 5 ） ,and the peptides with Mr 〉400 D were about 73% in all the enzymolysis solutions. Conclusion The technique is simple and reproducible for the preparation of heparolysate.