发酵类抗生素中残留蛋白通用性测定方法的探讨

发布时间：2019-02-26 20:53

【摘要】：目的:本研究探讨并建立了采用凝胶过滤色谱和Bradford法测定发酵类抗生素克拉维酸钾、硫酸链霉素和硫酸卷曲霉素中残留蛋白的分析方法。方法:采用凝胶过滤色谱先将蛋白与抗生素分离,富集蛋白组分,再冻干、复溶,最后利用Bradford法(考马斯亮蓝染色法)对残留蛋白进行测定。分别采用SephadexTMG-15(300 mm×15 mm,40~120μm)和SephadexTMG-25(100 mm×26 mm,90μm)色谱柱,以水为流动相,检测波长为280 nm,柱温30℃,进样量为500μL。结果:凝胶过滤法收集蛋白组分方法平均回收率约为97.4%,Bradford法的蛋白浓度在2~40μg·m L-1(r=0.999 6)内线性关系良好,蛋白(BSA)平均回收率约为95.4%,Bradford法的精密度小于3.0%,最低检测限为0.2μg·m L-1。所建立方法的合成标准不确定度为0.001%,扩展不确定度为0.002%。结论:该方法准确、可靠,重复性好,可用于发酵类抗生素中残留蛋白的检测和控制。
[Abstract]:Aim: to study and establish a method for the determination of residual proteins in fermentative antibiotics potassium clavulanate, streptomycin sulfate and caprythromycin sulfate by gel filtration chromatography and Bradford. Methods: the protein was separated from antibiotics by gel filtration chromatography, then the protein components were enriched, then freeze-dried and redissolved. Finally, the residual protein was determined by Bradford method (Coomassie brilliant blue staining). SephadexTMG-15 (300 mm 脳 15 mm,40~ 120 渭 m) and SephadexTMG-25 (100 mm 脳 26 mm, 90 渭 m) column were used, respectively. The mobile phase was water, the detection wavelength was 280nm, column temperature at 30 鈩，

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