摘要
该文研究了中心组合设计和响应面分析优化猪肺肝素钠的提取工艺,通过D-254树脂交换吸附、沉淀剂除蛋白质、乙醇沉淀以及DEAE-Sephasoe fast flow离子交换柱层析纯化得到高纯度的猪肺肝素钠,最后采用琼脂糖凝胶电泳、红外光谱、紫外光谱和核磁共振波谱等对其进行初步鉴定。猪肺肝素钠的最佳提取工艺参数为:提取温度53℃,时间4 h,pH=8.5。粗品和纯化后产品的提取率分别为0.81%和0.05%,羊血浆法测定粗品和纯化后产品效价分别为6.5 U/mg和108.3 U/mg。红外光谱显示,纯化后样品在890 cm-1和940 cm-1处出现肝素钠特征吸收峰,紫外光谱最大吸收波长为205 nm,结合琼脂糖凝胶电泳和核磁共振波谱证明用上述方法制备的纯化后的样品为肝素钠。
In order to optimize the extraction process of heparin sodium and analyze its structure, this article used central composite design and response surface to optimize the process of enzymatic extraction of heparin sodium. The heparin sodium with high purity was obtained after the resinous adsorption, deproteinization by precipitator, alcohol precipitation and column chromatography for further isolation and purification. Agarose gel electrophoresis, infrared spectroscopy （ IR）, ultra violet （UV） spectrum and nuclear magnetic resonance （NMR） were used to elementarily identify the structure of heparin sodium. The optimum extraction conditions are extraction temperature 53 ℃, extraction time 4 h and pH = 8.5. The crude and purified product extraction yields are 0. 81% and 0. 05%, and the rough and depurated goods potency is 6. 5 U/mg and 108.3 U/rag by the method of ovine plasma, respectively. An IR spectrum shows the characteristic absorption peaks are 890 cm^-1 and 940 cm^-1, and UV spectrum displays the maximum absorption wavelength is 205 nm. Based on the analyses of both agarose gel eleetrophoresis and NMR, the rarefied product is identified as heparin sodium.
出处
《精细化工》
EI
CAS
CSCD
北大核心
2014年第5期597-602,共6页
Fine Chemicals
基金
四川省科技厅项目(11ZS2013)~~
关键词
肝素钠
分离
结构
响应面分析
催化与分离提纯技术
heparin sodium
separation
structure
response surface methodology
catalysis, separationand purification technology
