利用免疫亲和色谱柱特异性敲除甘草酸的方法学研究

Methodology on specific knock out of glycyrrhizic acid by immunoaffinity chromatography column

目的制备特异性敲除甘草酸的免疫亲和色谱柱。方法抗甘草酸单克隆抗体与CNBr活化的Sepharose 4BTM凝胶偶联制备甘草酸特异性敲除免疫亲和色谱柱,利用HPLC法测定敲除液与洗脱液中甘草酸质量浓度,考察甘草酸免疫亲和色谱柱的最大载样量、敲出率、精密度、准确度及稳定性等参数。结果甘草酸免疫亲和色谱柱的最大载样量为1.326 11 mg;日内及日间精密度总RSD分别为0.65%,相对误差(RE)为0.37%;免疫亲和色谱柱稳定性实验显示32 d内甘草酸敲除率的RSD为1.37%,稳定性良好。结论制备的甘草酸特异性敲除免疫亲和色谱柱能快速、高效、稳定地敲除甘草酸。

Objective To prepare the glycyrrhizic acid （GA） imrnunoaffinity chromatography column, which could specifically knock out the GA. Methods An immunoaffinity chromatography （IAC） column was developed by covalently coupling the anti-GA-MAh to CNBr-activated SepharoseTM 4B. The concentration of GA was detected by HPLC, and the maximum coupling capacity, stability, precision, and accuracy of the IAC column for GA were studied. Results The maximum capacity of the IAC column for GA was 1.326 11 mg, the precision （RSD） of GA was 0.65%, and the accuracy （RE） was 0.37%. RSD of GA stayed 1.37% during 32 d, which showed a pretty well stability. Conclusion GA-IAC column could rapidly, effectively, and stably knock out the GA.