威廉环毛蚓提取物蛋白质含量测定方法研究
Determination of protein content in the extract from Pheretima guillelmi
蒋巧萍 1宇泉霖 1宋嘉佳 1李寒冰 1李敏枫 1檀世祖 1王天舒 1蔡程科1
作者信息
- 1. 北京中医药大学中药学院,北京 102488
- 折叠
摘要
目的 建立威廉环毛蚓提取物蛋白质含量测定的最佳方法.方法 以氨基酸分析仪测定的威廉环毛蚓提取物中的氨基酸含量为指标,对蛋白质含量检测方法Lowry法2、改良Lowry法、BCA法、Bradford法进行对比评价.结果 Lowry法2、改良Lowry法、BCA法、Bradford法测定威廉环毛蚓提取物蛋白质含量(上清液)结果分别为27.03％、45.85％、31.58％、11.11％,威廉环毛蚓提取物蛋白质含量(混悬液)的结果分别为38.44％、52.39％、44.06％、11.56％.采用氨基酸分析仪法测定得到威廉环毛蚓提取物中游离氨基酸含量为4.18％,将威廉环毛蚓提取物蛋白质酸水解之后测定总氨基酸含量为48.53％.结论 在威廉环毛蚓提取物样品溶液的处理方式上,与样品上清液相比,样品混悬液的测定结果更接近样品中蛋白质的真实含量.以牛血清白蛋白(BSA)为对照品,相较于Lowry法2、改良Lowry法和Bradford法,BCA法更适宜用于威廉环毛蚓提取物中的蛋白质含量测定.
Abstract
Objective To establish an optimal method determine the protein content in the extract from Pheretima guillelmi.Methods Amino acid content of the extracts from Pheretima guillelmi determined by amino acid analyzer was used as an index for the comparative evaluation of protein content assay methods namely Lowry's method 2,modified Lowry's method,BCA method and Bradford method.Results The protein content(supernatant)by Lowry's method 2,modified Lowry's method,BCA method,and Bradford's method was 27.03％,45.85％,31.58％,and 11.11％,and that of suspension was 38.44％,52.39％,44.06％,and 11.56％,respectively.We used the amino acid analyzer to obtain a free amino acid content of 4.18％,and the total amino acid content was 48.53％after acid hydrolysis of the protein.Conclusion Considering the way the sample solution is handled,the sample suspension is closer to the real content of protein in the sample as compared to the sample supernatant.Bovine albumin(BSA)was used as a control,the BCA method is more suitable for the determination of protein content in the extract of Pheretima guillelmi as compared to Lowry's method 2,modified Lowry's method and Bradford method.
关键词
威廉环毛蚓/蛋白质/Lowry法2/改良Lowry法/BCA法/Bradford法/氨基酸自动分析仪Key words
Pheretima guillelmi/protein/Lowry's method 2/modified Lowry's method/BCA method/Bradford method/amino acid auto analyzer引用本文复制引用
基金项目
国家科技重大专项研发项目(2011ZX09201-201-15)
北京市科技计划专项研发项目(Z16010101390)
出版年
2024
NETL
NSTL
万方数据