反相高效液相色谱法测定抗生素AGPM发酵液中的主要成分

Determination of AGPM Antibiotics in Fermentation Broth by Reversed-Phase High Performance Liquid Chromatography

主要探讨了高效液相色谱法(HPLC)测定新型抗生素AGPM发酵液中主要成分的可行性,优化了反相高效液相色谱法分析发酵液组分的色谱条件。在WatersTMNova PakR ○C18色谱柱上,以甲醇 0 01mol/L乙酸铵溶液(pH5 20)为流动相,其中甲醇的体积分数按线性梯度方式在60min内从30%升至50%,流动相流速为1 0mL/min,在330nm处进行紫外检测,发酵液中各主要组分均能很好地分离。采用该方法对AGPM发酵液中5种主要组分的线性范围、检出限、相对标准偏差和回收率分析考察,结果表明其灵敏度高、定量准确、重现性好,适合于发酵液这种复杂体系中AGPM等主要组分的分离测定。

A reversed-phase high performance liquid chromatography was developed to assay main components in anti-gram positive microorganism (AGPM, a novel antitumor an tibiotics) fermentation broths. The AGPM antibiotics were successfully separated on a WatersTM Nova-PakR○C18 column (150 mm×39 mm id, 5 μm) u sing a mixture of methanol-001 mol/L ammonium acetate (pH 520) solution as mobile phase under gradient elution at a flow rate of 10 mL/min at amb ient temperature, and detected by a UV detector at 330 nm. The methanol volume f raction in the mobile phase increased from 30% to 50% in 60 minutes. The linear ranges, detection limits, RSDs and recoveries of five main components were evalu ated respectively. The results showed that the method is sensitive, accurate and suitable for analysis of AGPM antibiotics in complex mixtures, such as fermenta tion broth.