双液相萃取-离子交换层析法纯化猪硫酸软骨素  

Purification of Porcine Chondroitin Sulfate with Double-Liquid Extraction-Ion-Exchange Chromatography

作　　者：陆钊[1] 潘淑霞[2] 董树国[1] 曹宏梅[3] 赖红伟[1] 勾龙啸[4] 

机构地区：[1]吉林医药学院药学院化学教研室,吉林吉林132013 [2]吉林医药学院生物医学工程系数学教研室,吉林吉林132013 [3]吉林医药学院实验中心基础化学实验室,吉林吉林132013 [4]吉林医药学院药学院08级临床药学本科班,吉林吉林132013

出　　处：《西南大学学报（自然科学版）》2012年第7期136-140,共5页Journal of Southwest University(Natural Science Edition)

基　　金：吉林市科技局科研课题(200903016)

摘　　要：采用聚乙二醇(PEG)/硫酸铵双液相萃取-离子层析法纯化猪硫酸软骨素,红外光谱法测定结构,咔唑分光光度法测定含量.以PEG的相对分子质量和质量分数、(NH4)2SO4的质量分数、pH值和萃取次数为考察因素,以猪硫酸软骨素的回收率和纯度为考察指标,正交设计法优化双液相萃取条件.IRC50离子交换树脂对萃取液纯化,红外光谱法和咔唑分光光度法鉴定纯化后猪硫酸软骨素的含量和结构.最佳纯化条件为:PEG相对分子量为4 000,质量分数为30%,(NH4)2SO4的质量分数为26%,pH值为10,萃取3次.萃取样品经IRC50离子交换树脂纯化后,质量浓度在0～250mg/L范围内线性关系良好,相关系数r=0.999 7,样品含量为90.49%,相对标准偏差为0.58%.红外吸收光谱比较纯化后猪硫酸软骨素和标准品硫酸软骨素-A,其特征吸收峰均在3 400,1 620,1 560,1 240,860cm-1处.该方法精确度和准确度都较高,可用于猪硫酸软骨素的纯化.Purpose： To purify porcine chondroitin sulfate with polyethylene glycol ammonium sulfate double-liquid extraction-ion-exchange chromatography, determine its structure with infrared spectroscopy and determine its con- tent with carbazole spectrophotometry. Methods.. With the relative molecular mass and mass fraction of polyethylene glycol （PEG）, the mass fraction of ammonium sulfate, pH and extraction times as the factors to be examined, the conditions of double-liquid extraction were optimized with the orthogonal design, the extract was purified with IRC50 ion exchange resins, its content was determined with carbazole and its structure was examined with spectrophotometry. Results： The best extraction conditions were as follows： relative molecular mass and mass fraction of PEG being 4 000 and 30%, mass fraction of （NH4）2SO4 being 26% , pH being 10 and extraction being made 3 times. The mass concentration of extraction samples purified with IRC50 ion exchange resins was in a good linear relationship within 0--250 mg/L, with a correlation coefficient of 0. 999 7. The content of the sample was 90.49o./oo and RSD=0.58G. The infrared absorption spectra of the purified porcine chondroitin sulfate and the standard chondroitin sulfate-A showed that both had their characteristic absorption peaks at 3 400, 1 620, 1 560, 1 240 and 860 cm 1. Conclusion： This method is precise and accurate and can be used to purify porcine chondroitin sulfate.

关 键 词：猪硫酸软骨素 双液相萃取法 离子交换层析法 红外光谱法 咔唑分光光度法 

分 类 号：S828[农业科学—畜牧学]