用HPLC-ESI-IT-MS^n法研究庆大霉素C_(1a)的杂质谱  

Study on Impurity Profile of Gentamicin C_(1a) by HPLC-ESI-IT-MS^n

作　　者：杨昊[1] 袁耀佐[1] 赵恂[1] 谭力[1] 张玫[1] 钱文[1] 

机构地区：[1]江苏省食品药品检验所,南京210009

出　　处：《药学与临床研究》2012年第5期413-418,共6页Pharmaceutical and Clinical Research

摘　　要：目的:建立用于研究庆大霉素C1a杂质谱的HPLC-ESI-IT-MSn方法,并对检出杂质结构进行快速推定。方法:用宽pH范围的Gemini NX C18(4.6 mm×150 mm,5μm)色谱柱,以[水-氨水-冰醋酸(96∶3.6∶0.4)]-甲醇(85∶15)为流动相,用ESI-IT-MSn检测(质谱条件:ESI离子源,离子源温度350℃,雾化室压力275.8 kPa,干燥气流速10 L.min-1,正离子检测方式)。结果:该色谱条件能将庆大霉素C1a与主要杂质分离,从庆大霉素C1a原料中检出13个杂质,对其中12个的结构进行了归属。通过与杂质对照品的色谱保留时间,一、二级质谱比较,对已知杂质进行了鉴定;借助总结出来的氨基糖苷类抗生素的特征裂解碎片离子和质谱裂解规律,根据一级、二级质谱或多级质谱信息,对无杂质对照品的未知杂质结构进行推导,结果显示,庆大霉素C1a样品中含有庆大霉素A、庆大霉素B、西索米星、小诺霉素、威达米星或它们的同分异构体和同系物,并含有庆大霉素C1a或上述杂质的降解产物。结论:该方法可以用于庆大霉素C1a的工艺优化和质量控制。Objective： An HPLC-ESI-IT-MSn method was developed for the study of impurity profile and rapid characterization of the impurities detected in gentamicin C1a samples.Methods： The separation was performed on a broad pH value column（Gemini NX C18,4.6 mm×150 mm,5 μm） using [water-ammonia-glacial acetic acid（96∶3.6∶0.4）]-methanol（85∶15） as mobile phase.The temperature of ESI ion source was 350℃,the nebulizing pressure was 275.8 kPa,the dry gas flow was 10 L·min-1 and the analytes were detected in a positive mode.Results： The method showed good separations among gentamicin C1a and its main impurities.A total of 13 impurities were detected in a gentamicin C1a sample and 12 were deduced.Some known impurities were identified by comparing their retention times,MS and MS2~n information with those of the reference substances.The structures of the other unknown compounds were deduced based on comparison of fragmentation patterns with those of the reference substances investigated in LC/MSn experiments by the use of electrospray ion trap mass spectrometry before.The impurities detected in a gentamicin C1a sample were identified as gentamicin A,gentamicin B,sisomicin,micronomicin,verdamicin and their isomeric compounds,homologs or degradation products.Results： The method established could be applied to the optimization of manufacture process and the quality control of gentamicin C1a.

关 键 词：庆大霉素C1a 杂质谱 高效液相色谱-电喷雾-离子阱质谱 结构推定 

分 类 号：R927[医药卫生—药学]