应用羧甲基纤维素法从猪血球中提取卟啉铁的方法研究
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摘要
本论文主要研究了猪血球的实验室破壁方法、测定血球破壁率的方法和应用羧甲基纤维素(CMC)从猪血球中提取卟啉铁的工艺参数,并对产品的纯度、产量、收率、溶解度和分离出的珠蛋白蛋白含量进行了分析。探讨了卟啉铁产品的开发利用前景,为提取卟啉铁的工业化生产提供了理论依据。主要研究结果如下:
1.改进了实验室血球细胞破壁方法,具体工艺为:血球细胞中加入5倍去离子水,0.2倍乙醇,机械搅拌30min。
2.确定了用标准曲线法测定血球细胞破壁率的方法。
3.改进了应用CMC从猪血球中提取卟啉铁的工艺流程。
4.确定了应用CMC法提取卟啉铁的最佳工艺参数,具体为:分离珠蛋白与卟啉铁的最佳pH值为1.7、分离搅拌时间为2h、吸附pH值为3.2、CMC添加量为血球的0.3倍、吸附搅拌时间为45min、提取pH为4.8、提取搅拌时间为70min。
5.试验分析结果为:提取得粗卟啉铁的纯度在73%左右;产量在2.36g(每200mL血球的所得提取量)左右;收率在90%左右;其珠蛋白的蛋白含量在92%左右;卟啉铁在0.1mol/L的NaOH溶液,3%的Na_2CO_3和NaHCO_3溶液中的溶解度分别为98.91%,98.11%和94.73%;且每g卟啉铁的生产成本仅为0.4元。
This paper studied on the method of disrupting red cell wall in the laboratory condition,the assaying method of broken-red-cell rate and the technological procedure parameters of using Carboxym Mthyl Cellulose method to extract Iron-porphyrin from the porcine red blood cell,in the mean time analyzed the Iron-porphyrin purity, production, yield,dissolution rate and the crude protein content of globin. Then this paper was intended to introduce the utilization of Iron-porphyrin and it supplied theoretical basis to industrial production of Iron-porphyrin. The results were as follow:
1. The method of disrupting red cell in the laboratory was improved. The results showed that the optimum conditions are: 1 times blood cell,add 5 times water and 0.2 times ethanol,and then give mechanical churn for 30 minutes.
2. The standard curve method was determined to assay the broken-red-cell rate.
3. The procedure of extracting Iron-porphyrin from porcine blood corpuscle by using Carboxym Ethyl Cellulose method was improved.
4. The optimum conditions for the extraction techniques of Iron-porphyrin were: separating Iron-porphyrin from globin at pH1.7,at about 2 hours’churning time for sepration,at pH 3.2 for absorbing Iron-porphyrin,at 0.3 times CMC supplement,at about 45 minutes’churning time for absorbtion,the extraction pH at 4.8,70 minutes’for churning.
5. The content of Iron-porphyrin was about 73%without further purification. The yield was about 2.36g,the dissolution rate of Iron-porphyrin in the 0.1mol/L NaOH,3% Na2CO3 and NaHCO3 was 98.91%,98.11%and 94.73%respectively. The crude protein content of globin was about 92%,and it cost 0.4 RMB for per gram extractives.
1.改进了实验室血球细胞破壁方法,具体工艺为:血球细胞中加入5倍去离子水,0.2倍乙醇,机械搅拌30min。
2.确定了用标准曲线法测定血球细胞破壁率的方法。
3.改进了应用CMC从猪血球中提取卟啉铁的工艺流程。
4.确定了应用CMC法提取卟啉铁的最佳工艺参数,具体为:分离珠蛋白与卟啉铁的最佳pH值为1.7、分离搅拌时间为2h、吸附pH值为3.2、CMC添加量为血球的0.3倍、吸附搅拌时间为45min、提取pH为4.8、提取搅拌时间为70min。
5.试验分析结果为:提取得粗卟啉铁的纯度在73%左右;产量在2.36g(每200mL血球的所得提取量)左右;收率在90%左右;其珠蛋白的蛋白含量在92%左右;卟啉铁在0.1mol/L的NaOH溶液,3%的Na_2CO_3和NaHCO_3溶液中的溶解度分别为98.91%,98.11%和94.73%;且每g卟啉铁的生产成本仅为0.4元。
This paper studied on the method of disrupting red cell wall in the laboratory condition,the assaying method of broken-red-cell rate and the technological procedure parameters of using Carboxym Mthyl Cellulose method to extract Iron-porphyrin from the porcine red blood cell,in the mean time analyzed the Iron-porphyrin purity, production, yield,dissolution rate and the crude protein content of globin. Then this paper was intended to introduce the utilization of Iron-porphyrin and it supplied theoretical basis to industrial production of Iron-porphyrin. The results were as follow:
1. The method of disrupting red cell in the laboratory was improved. The results showed that the optimum conditions are: 1 times blood cell,add 5 times water and 0.2 times ethanol,and then give mechanical churn for 30 minutes.
2. The standard curve method was determined to assay the broken-red-cell rate.
3. The procedure of extracting Iron-porphyrin from porcine blood corpuscle by using Carboxym Ethyl Cellulose method was improved.
4. The optimum conditions for the extraction techniques of Iron-porphyrin were: separating Iron-porphyrin from globin at pH1.7,at about 2 hours’churning time for sepration,at pH 3.2 for absorbing Iron-porphyrin,at 0.3 times CMC supplement,at about 45 minutes’churning time for absorbtion,the extraction pH at 4.8,70 minutes’for churning.
5. The content of Iron-porphyrin was about 73%without further purification. The yield was about 2.36g,the dissolution rate of Iron-porphyrin in the 0.1mol/L NaOH,3% Na2CO3 and NaHCO3 was 98.91%,98.11%and 94.73%respectively. The crude protein content of globin was about 92%,and it cost 0.4 RMB for per gram extractives.
引文
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