摘要
介绍草乌毒蛋白的提取方法。草乌根经pH7.2的PBS溶液 (含9g/LNaCl)浸提,浸提液经CM -SFF柱和SephacrylS -200凝胶过滤柱分离,然后在高效凝胶过滤柱上制备ACO毒蛋白,利用光电二极管阵列检测器的光谱性能确认色谱峰的纯度,并根据标准蛋白的相对分子质量校正曲线求得ACO的相对分子质量。
The isolation and purification of a toxic protein from the root of the plant Aconitum kusnezoffii Reichb(″ Caowu″ in Chinese) is described. The crude toxin was extracted from the plant roots by dialysis with a PBS solution of 9 g/L NaCl at pH 7.2. Separation of the extract was obtained on a CM-SFF column with a phosphate buffer gradient elution. The collected solution for the first peak was then separated on a gel _ filtration Sephacryl S-200 column, using neutral phosphate as the mobile phase. The protein was further purified on two Protein-pak 125 columns connected in series, using a mobile phase of 0.2 mol/L phosphate buffer(pH 6.5). The protein gives a typical absorption spectrum of protein. The purity of the protein peak can be judged from the spectrum, using a photodiode array detector. The re_lative molecular mass of this protein measured by two protein columns connected in series is ca. 31 500. The amino acid composition of this protein was determined by OPA post _ column derivatization-fluorescence detection.
出处
《分析测试学报》
CAS
CSCD
2000年第3期40-42,共3页
Journal of Instrumental Analysis
关键词
草乌
毒蛋白
氨基酸
有毒物质
分离
性质
Radix aconiti Kusnezoffii
Protein
Relative molecular mass
Amino acid
Toxin
