高压液相色谱技术分离制备杆菌肽各组分的研究

目的 开发制备型高压液相色谱分离杆菌肽各组分的方法.方法 根据杆菌肽各组分化学结构性质,采用纳微UnisilC1s柱,通过改变流动相pH和流动相配比,以及不同的上样量,探索杆菌肽各组分分离的方法.收集分离得到的不同组分,经脱盐-冷冻干燥,得到相应杆菌肽组分冻干粉.结果 纳微Unisil C18柱可以用于杆菌肽各组分的分离,流动相配比为甲醇(A)∶1.54‰乙酸铵水溶液(乙酸调pH至5.0)-乙腈(9∶1,V/V)] (B)=9.0∶ 11.0(V/V),总流速40 mL· min-1,上样量200mg,检测波长为254 nm.制备得到的杆菌肽各组分纯度均大于95％.结论 该方法上样量大,稳定性好,收集到的各组分纯度高,为杆菌肽各组分的分离制备提供了借鉴.

Preparation of bacitracin components by preparative HPLC

Objective To develop the separation and preparation method of bacitracin components by preparative HPLC.Methods The appropriate separation method was determined through the pH of mobile phase,the ratio of mobile phase,and the loading quantity of sample by using nanomicro Unisil C1s.Freeze-dried powders of bacitracin components were obtained by desalination and freeze drying of the collected solution.Results Nanomicro Unisil C1s chromatographic column could be used for separation.The ratio of mobile phase was A∶ B =9.0∶11.0 (V/V) A was methanol and B consisted of 1.54‰ ammonium acetate solution (pH adjusted to 5.0)-acetonitrile (9∶ 1,V/V)].The flow rate was 40 mL · min-1 and the sample load was 200 mg.The wavelength was 254 nm.The purities of bacitracin components were all above 95％.Conclusions The method is of great loading quantity of sample,good stability,and high purity of the prepared components.It provides reference for the separation and preparation of bacitracin components.