猪肺肝素钠提取工艺优化、分离与初步鉴定

Separation, Purification and Identification of Porcine Pulmonary

摘要:

本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物化和仪器分析对其进行初步鉴定。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为别为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。

Abstract:

In order to optimize the extraction process of heparin sodium and analysis the structure. This article used central composite design and response surface to optimize the extraction process of heparin sodium’s enzymatic method. The heparin sodium with high purity was obtained after the resinous adsorption, precipitator for deproteinization, 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) used to identify the structure of heparin sodium elementarily. The optimum extraction conditions are extraction temperature 53 ℃, extraction time 4h and pH 8.5. The crude and purified product extraction yield are 0.81 % and 0.05 %, the rough and depurated goods potency are 6.5 U/mg and 108.3 U/mg by the method of ovine plasma, respectively. An IR spectrum shows the characteristic absorption peaks are 890 cm-1 and 940 cm-1, UV spectrum displays the maximum absorption wavelength is 205 nm. Combined with agarose gel electrophoresis and NMR, the rarefied product is identified as heparin sodium.