【摘要】：目的:本研究探討并建立了采用凝膠過濾色譜和Bradford法測定發(fā)酵類抗生素克拉維酸鉀、硫酸鏈霉素和硫酸卷曲霉素中殘留蛋白的分析方法。方法:采用凝膠過濾色譜先將蛋白與抗生素分離,富集蛋白組分,再凍干、復(fù)溶,最后利用Bradford法(考馬斯亮藍(lán)染色法)對殘留蛋白進(jìn)行測定。分別采用SephadexTMG-15(300 mm×15 mm,40~120μm)和SephadexTMG-25(100 mm×26 mm,90μm)色譜柱,以水為流動(dòng)相,檢測波長為280 nm,柱溫30℃,進(jìn)樣量為500μL。結(jié)果:凝膠過濾法收集蛋白組分方法平均回收率約為97.4%,Bradford法的蛋白濃度在2~40μg·m L-1(r=0.999 6)內(nèi)線性關(guān)系良好,蛋白(BSA)平均回收率約為95.4%,Bradford法的精密度小于3.0%,最低檢測限為0.2μg·m L-1。所建立方法的合成標(biāo)準(zhǔn)不確定度為0.001%,擴(kuò)展不確定度為0.002%。結(jié)論:該方法準(zhǔn)確、可靠,重復(fù)性好,可用于發(fā)酵類抗生素中殘留蛋白的檢測和控制。
[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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