乙酰螺旋霉素组分分析中色谱柱的理性选择方法探讨(英文)

Facilitated column selection in the separation of acetylspiramycin components using a simple column classification system

乙酰螺旋霉素为碱性的多组分抗生素,是以螺旋霉素为起始原料的半合成抗生素。乙酰螺旋霉素原料药中除了单-乙酰螺旋霉素Ⅱ和Ⅲ、双-乙酰螺旋霉素Ⅱ和Ⅲ四种主成分外,还能检出70多种小组分,这些小组分的结构与主成分相似或为主成分的同分异构体,故将乙酰螺旋霉素的各组分基线分离很具挑战性;在实际工作中还发现,采用不同品牌的C_(18)柱所得的乙酰螺旋霉素组分测定结果有时差异较大,有必要采用科学的方式理性挑选适宜的色谱柱进行其组分测定。本文利用国际上较有影响力的鲁汶大学所建的色谱柱分类系统数据库,以文献报道的CAPCELL PAK MG C_(18)柱(250 mm×4.6 mm,5μm)作为参比柱,选择了3组选择性不同的色谱柱,并按照中国药典2010版乙酰螺旋霉素各论项下的方法考察了这3组色谱柱的实际分离情况。与参比柱的结果相比,F<2组的分离效果最好,F>6组的分离效果最差,而2<F<6组的分离效果介于两者之间,与色谱柱分组结果呈较好的相关性;而且,F<2组和2<F<6组色谱柱(F≤2.7)的组分测定结果与参比柱结果之间的绝对差异均未超过2.0%,而F>6组色谱柱(F≥9.9)的绝对差异则超过2.5%,甚至高达8.2%,进一步验证了该色谱柱分类系统在适宜色谱柱选择方面的作用。

Acetylspiramycin（ASPM）,a 16-membered basic macrolide antibiotic,is the acetylated derivative of spiramycin.In addition to the four main components,more than seventy minor components could be present in ASPM.Thus,it is a challenge to obtain a baseline separation of ASPM components.Meanwhile,in some cases it was found that the results obtained by different brands of C_（18） columns were significantly different,indicating the necessity of a rational column selection for the separation of ASPM components.In this paper,we attempted to facilitate column selection for the analysis of ASPM using the database of the column characterization system established by Leuven University.With the CAPCELL PAK MG C_（18）（250 mm×4.6 mm,5μm） as the reference column,three groups of columns（F2 group：similar selectivity;2F6：intermediate ranked group;F6 group： different selectivity,compared to the reference column） were selected,and their performances in the separation of ASPM components were evaluated under the chromatographic conditions described in Chinese Pharmacopoeia monograph.A good relationship was demonstrated between the ranking of columns and their selectivity in the separation of ASPM components,indicating the column classification system established by Leuven University was a helpful tool in the selection of suitable columns for the analysis of ASPM,a complex mixture of basic compounds.