【作者】 袁耀佐； 谭力； 张玫； 钱文； 张正行；

【Author】 Yuan Yao-zuo~1,Tan Li~1,Zhang Mei~(1*),Qian Wen~1 and Zhang Zheng-xing~2 (1.Jiangsu Institute for drug control,Nanjing 210008) (2.Department of Pharmaceutical Analysis,China Pharmaceutical University,Nanjing 210009)

【机构】 江苏省药品检验所； 中国药科大学药物分析教研室；

【摘要】 目的建立鉴定硫酸依替米星的HPLC-ESI-MS~2分析方法,对硫酸依替米星成品中的有关物质进行鉴定。方法利用宽pH范围的Gemini NX C18(4.6mm×150mm,5μm)色谱柱,在碱性条件下,以[水-氨水-冰醋酸(96:3.6:0.4)]-甲醇(70:30)为流动相,直接分离各组分;并用HPLC-ESI-MS~2检测样品中各组分,质谱条件:ESI离子源,离子源温度350□,雾化室压力50 psi,干燥气流速10 L/min,正离子检测方式;对有对照品的有关物质采用比对有关物质与对照品的色谱保留时间、准分子离子的质荷比及二级质谱方法进行鉴定,无对照品的有关物质,则是先通过解析庆大霉素Cla、小诺霉素、依替米星的二级质谱,对该类化合物的二级质谱裂解规律进行归纳,并以此推测未知新化合物进行结构,再用推测的结构对二级质谱进行解析,如所有的碎片离子均合理解释,表明推测合理。结果用建立新方法,从硫酸依替米星成品中主要检出19个有关物质,对其中3个为已知化合物,即庆大霉素Cla、小诺霉素和奈替米星和1个依替米星异构体进行了确证,另外10个有关物质的结构除部分取代基的取代未知不能肯定外也基本得到鉴定,其余5个有关物质提供了分子量和二级质谱信息;结果显示,硫酸依替米星样品中,有4个依替米星同分异构体、4个小诺霉素同分异构体、2个依替米星降解物、2个依替米星同系物。结论新建立的HPLC-ESI-MS~2方法可用于该品种的有关物质的结构鉴定和定性分析,该方法为依替米星质量控制和工艺优化研究提供了可靠的分析手段。更多还原

【Abstract】 Objectives To establish a HPLC-ESI-MS² method for the identification of the related substances in etimicinsulfate.Methods by using a broad pH range column（Gemini NX C18,4.6mm×150mm,5μm）,basic mobile phase which was[H₂O-ammonia-glacial acetic acid] （96:3.6:0.4） -methanol,the related substances in etimicinsulfate were separated and detected by electrospray ionization and positive ion monitoring,the ESI ion source condition was following: temperature is 350℃,nebulizing pressure is 50 psi,dry gas flow is 10 L /min.some related substances in etimicinsulfate were identified by comparing the retention time in chromatography, [M+H]⁺ spectrum and MS² spectrum with reference substances’,the others which haven’t reference substances were identified or speculated by analyzing their MS² fragmentation with the help of a rule summarized from the MS² fragmentation of gentamicin C1a,micronomicin and etimicin.Results nineteen related substances in etimicinsulfate were separated and detected. Four related substances were identified as gentamicin C1a,micronomicin,netimicin and one of etimicin’s isomeric compounds,the other ten related substances which were not reported previously were speculated and identified except some substituent on them can’t be determinated, and the other five related substances can not be identified but provided[M+H]⁺ spectrum and MS² spectrum.Among them,there four etimicin’s isomeric compounds,four micronomicin’s isomeric compounds,two degradation products of etimicin and two homolog of etimicin,etc.Conclusions the method could be used in identification and analyzing qualitatively the related substances in etimicinsulfate and quality control and optimization of etimicin’ synthetic technology.更多还原