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肝水解肽的制备及相对分子质量分布研究
引用本文:蒋桂香,李延平,刘中原,吴科锋,刘义,李立.肝水解肽的制备及相对分子质量分布研究[J].中国生化药物杂志,2012,33(5):603-606.
作者姓名:蒋桂香  李延平  刘中原  吴科锋  刘义  李立
作者单位:1. 广东双林生物制药有限公司湛江市生物制药工程中心,广东湛江,524005
2. 广东医学院广东天然药物研究与开发重点实验室,广东湛江,524023
基金项目:东莞市高等院校科研机构和医疗卫生单位科技计划项目(2008108101059)
摘    要: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。

关 键 词:肝水解肽  酶解  高效液相色谱法  相对分子质量  分布

Study on preparation method and molecular weight distribution of polypeptide in heparolysate
JIANG Gui-xiang , LI Yan-ping , LIU Zhong-yuan , WU Ke-feng , LIU Yi , LI Li.Study on preparation method and molecular weight distribution of polypeptide in heparolysate[J].Chinese Journal of Biochemical Pharmaceutics,2012,33(5):603-606.
Authors:JIANG Gui-xiang  LI Yan-ping  LIU Zhong-yuan  WU Ke-feng  LIU Yi  LI Li
Institution:1.Guangdong Shuanglin Biopharmaceatical Co.,Ltd.,the Engineering Center of Zhanjiang Biopharmacy,Zhanjang 524005,China;2.Guangdong Medical College,Guangdong Key Laboratory for Research andDevelopment of Natural Drugs,Zhanjiang 524023,China)
Abstract: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 prepared 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 a good linear 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.
Keywords:heparolysate  enzymolysis  HPLC  molecular weight  distribution
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