白及溶胶中植物蛋白的定性定量方法研究  

Study on the qualitative and quantitative methods for plant protein in Rhizoma Bletillae sol

作　　者：车向前[1] 陈芳 常明泉 陈林 CHE Xiang-qian;CHEN Fang;CHANG Ming-quan;CHEN Lin(Hubei Fangxian People,sHospital,Shiyan 442100,China;Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]湖北房县人民医院,湖北十堰442100 [2]十堰市太和医院(湖北医药学院附属医院),湖北十堰442000

出　　处：《实用药物与临床》2018年第7期815-818,共4页Practical Pharmacy and Clinical Remedies

基　　金：国家自然科学基金(81771522)

摘　　要：目的研究白及溶胶中植物蛋白定性定量方法。方法应用硝酸与蛋白质的硝化反应对其进行定性鉴别,应用双缩脲试剂与蛋白质的染色反应进行色谱鉴别,应用考马斯亮蓝染色法对其进行定量检测。结果白及溶胶加入硝酸后呈现蛋白质硝化的黄色反应,与对照品反应颜色一致;白及溶胶溶液与双缩脲试剂反应呈紫色,在540 nm处有最大吸收,与对照品一致;该植物蛋白样品溶液在595 nm处有最大吸收,标准蛋白溶液在3.92～19.61μg/ml范围内呈良好的线性关系,曲线方程为Y=0.036 X-0.000 5(r=0.999 5),平均回收率为96.2%,RSD为1.33%。结论硝酸用于白及溶胶中植物蛋白的定性鉴别,速度快,黄色呈现明显;考马斯亮蓝染色法用于白及溶胶中植物蛋白的含量测定,方法简便、快捷,灵敏度高,结果重现性好,适用于该溶胶的定量。Objective To study the qualitative and quantitative methods for plant protein in Rhizoma Bletillae sol. Methods Quantitative identification was conducted through the reaction of nitric acid and protein; chromatographic identification was conducted through the dyeing reaction between biuret reagents and protein; quantitative detection was conducted by the application of coomassie brilliant blue staining method. Results After adding nitric acid into Rhizoma Bletillae solution,the yellowreaction of protein nitration had the same color as that of reference substance reaction; the purple color was observed when Rhizoma Bletillae solution reacted with biuret reagents; maximum absorption was observed at 540 nm,which was the same as reference substance; the maximum absorption of the plant protein sample solution was observed at 595 nm; a good linear relationship of the standard protein solution was shown at the range of 3. 92 ～ 19. 61 μg/ml; the curve equation was Y = 0. 036 X-0. 000 5（r = 0. 999 5）; the average recovery was96. 2%,RSD 1. 33%. Conclusion Qualitative identification of plant protein in Rhizoma Bletillae sol by nitric acid is fast,and the phenomenon of color is obvious. Coomassie brillant blue staining method,used in content determination of plant protein in Rhizoma Bletillae sol,is simple and rapid,with high sensitivity,and the results are reproducible,which is suitable for quantitative identification of this sol.

关 键 词：白及溶胶 植物蛋白 考马斯亮蓝 定性 定量 

分 类 号：R286.0[医药卫生—中药学]