伊拉兔肺中硫酸软骨素C的分离纯化及二糖组成分析  

Isolation,purification and disaccharide composition analysis of chondroitin sulfate C from IRA rabbit lung

作　　者：郎银芝 王超 陈晨[1] 张晓[1] 刘潇潇[1] 单鑫迪[1] 蔡超[1] 于广利[1] 

机构地区：[1]山东省糖科学与糖工程重点实验室,海洋药物教育部重点实验室,中国海洋大学医药学院,青岛266003 [2]青岛市食品药品检验研究院,青岛266071

出　　处：《中国新药杂志》2016年第10期1165-1169,共5页Chinese Journal of New Drugs

基　　金：国家科技支撑计划(2013BAB01B02)

摘　　要：目的:从伊拉兔肺中提取分离硫酸软骨素C(chondroitin sulfate C,CSC),并对其进行二糖组成分析。方法:采用丙酮、氯仿-甲醇(2∶1)对兔肺进行脱脂,利用木瓜蛋白酶对脱脂粉进行酶解。提取的粗品经75%乙醇沉淀,氯代十六烷基吡啶(CPC)将其中的酸性多糖富集获得总糖胺聚糖TFH。采用Q-Sepharose Fast Flow树脂对TFH分级纯化获得TF-C。采用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生反相HPLC法分析单糖组成,利用硫酸软骨素酶ABC降解,SAX-HPLC法分析二糖组成。结果:所得TF-C不含蛋白质,主要由葡萄糖醛酸(Glc A)和氨基半乳糖(Gal N)组成,二糖组成以不饱和△UA-Gal NAc6S为主,含有少量的△UA-GalNAc4S。结论:首次从伊拉兔肺中分离获得低硫酸化高纯度CSC,为伊拉兔肺高值化产品的开发和利用提供了理论依据。Objective： To extract and isolate chondroitin sulfate C（ CSC） from IRA rabbit lung,and to analyze its disaccharide composition. Methods： The rabbit lung was defatted by acetone and chloroform-methanol（ 2∶ 1）,and digested by papain to release glycans. The extracted crude product was precipitated by 75% ethanol,and acidic polysaccharides were enriched by cetylpyridinium chloride（ CPC） to obtain the total glycosaminoglycans,TFH. TFH was fractionated by ion-exchange chromatography on Q-Sepharose Fast Flow column,and TF-C was acquired as a pure fraction. The monosaccharides hydrolyzed from TF-C were labeled with 1-phenyl-3-methyl-5-pyrazolone（ PMP） and analyzed by RP-HPLC. Subsequently,the disaccharide composition was determined by SAX-HPLC method after the digestion of TF-C with chondroitinase ABC. Results： TF-C mainly consisted of glucuronic acid（ Glc A） and galactosamine（ Gal N） while no protein was detected. The unsaturated disaccharides were mostly composed of △UA-GalNAc6 S with trace amount of △UA-GalNAc4 S. Conclusion： CSC with high purity is first time isolated from IRA rabbit lung,which could provide reference for the exploitation and utilization of high-valued products from IRA rabbit lung.

关 键 词：伊拉兔肺 硫酸软骨素C 分离纯化 二糖组成 

分 类 号：R931[医药卫生—生药学]