Avermectin粗粉结晶工艺探究及杂质分析.docVIP

Avermectin粗粉结晶工艺探究及杂质分析 　 作者：刘吉 季清荣 常志东，谢智 张贵峰 申淑锋 孙兴华 雷超 刘会洲 【关键词】; Avermectin；,高效液相色谱；,质谱；,杂质；,结晶 摘要： 采用高效液相色谱(HPLC)电喷雾串级质谱(ESIMSMS)的方法对avermectin粗粉中的杂质进行定性分析。确定了主要杂质成分为：与B1a结构相类似的B1b、A1a、A1b、A2a、A2b、B2a 6个组份，其中A1a、B1b、B2a多次结晶也不易去除。随后考察了avermectin粗粉在甲醇、乙醇、正丁醇和丙酮中的重结晶，甲醇和正丁醇对杂质的去除效果更好；乙醇与丙酮相当，但对杂质去除的选择性不同。　　关键词： Avermectin； 高效液相色谱； 质谱； 杂质； 结晶 ; Study on the crystallization of avermectin crude powder ; and its impurities analysis ABSTRACT Avermectin crude powder were purified by crystallization and its impurities were identified by HPLC (high performance liquid chromatography)ESIMSMS (electrospraymass spectrometrymass spectrometry). Main impurities in crude powder were B1b, A1a, A1b, A2a, A2b and B2a, which were similar to B1a in structure. The separation of A1a, B1b and B2a was very difficult due to their greatly similar structures to B1a. Recrystallizaton using methanol, ethanol, nbutanol and acetone as solvents were done respectively and it was found that methanol and nbutanol were good solvents for the separation of impurities. The impurities could be selectively separated using methanol or acetone.　KEY WORDS Avermectin; HPLC; Mass spectrometry; Impurities; Crystallization ; Avermectin是十六元大环内酯类抗生素，具有很强的杀虫活性，是一种良好的杀螨虫、昆虫和寄生虫的药物，可以用于人、畜和农作物［1］。作为杀虫剂，avermectin具有高效性和广谱性，它可以同时驱杀几乎所有的线虫类、昆虫类和螨虫类寄生虫，且一次用药能达到80%～100%的驱净率［2，3］。Avermectin的残毒低，有很好的杀虫选择性，是一种良好的大面积控制害虫的杀虫剂［4］，也是生产多种药物的原料，改造多产生在侧链，如C4”、C5和C25［5］。已商品化的衍生物有很多种，如：ivermectin(IVR)、emamectin(EMA)、doramectin(DOR)、moxidectin(MOX)、eprinomectin(EPR)等。Avermectin是由除虫链霉菌Streptomyces avermitilis产生的一组大环内酯类物质，包括结构相似的8种天然组分［6］。根据C5位上取代基不同，C22和C23之间的单双键差异及C25位上取代基的不同，分别用A、B；1、2；a、b组合表示(结构式见图1)，B组分药效较A组分强， B1组分较B2组分强， B1组分中Avermectins R1 R2 XYA1a CH3 C2H5 CH=CHA1b CH3 CH3 CH=CHB1a H C2H5 CH=CHB1b H CH3 CH=CHA2a CH3 C2H5 CH2CH(OH)A2b CH3 CH3 CH2CH(OH)B2a H C2H5 CH2CH(OH)B2b H CH3 CH2CH(OH)图1 天然Avermectins的化学结构［3］ 又以B1a药效更好一些［7］。所以avermectin的提纯一般针对B1a组分。Avermectin是胞内产物，其生产工艺包括发酵、浸提、浓缩结晶和重结晶提纯。其中结晶是avermectin纯化工艺的关键，目前工业上需多次