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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 this paper, the center combination design and response surface analysis were used to optimize the extraction process of pork lung heparin sodium. The purity of purified pork lung heparin sodium was optimized by D-254 resin exchange adsorption, precipitator deproteinization, ethanol precipitation and DEAE-Sepharose Fast flow ion exchange chromatography Of pig lung heparin sodium, and finally by agarose gel electrophoresis, infrared spectroscopy, ultraviolet spectroscopy and nuclear magnetic resonance spectroscopy for its initial identification. The optimal extraction parameters of pork lung heparin sodium were: extraction temperature 53 ℃, time 4 h, pH = 8.5. The crude and purified products were extracted at 0.81% and 0.05%, respectively, and the crude and purified titers of goat plasma were 6.5 U / mg and 108.3 U / mg, respectively. The infrared spectra showed that the purified heparin showed characteristic absorption peaks at 890 cm-1 and 940 cm-1, and the maximum absorption wavelength was 205 nm. The results of agarose gel electrophoresis and nuclear magnetic resonance spectroscopy The purified sample is heparin sodium.