依替米星有关物质的9-芴甲氧羰酰氯柱前衍生化高效液相色谱法检查与液质联用鉴定  

Determination and Identification of Related Substances of Etimicin Sulfate by HPLC and Liquid Chromatography Tandem Mass Spectrometry Using Pre-Column Derivatization with 9- Fluorenylmethyloxycarbonyl Chloride

作　　者：孙晶[1,2] 杭太俊[2] 袁耀佐[1] 谭力[1] 赵恂[1] 赵卫[1] 

机构地区：[1]江苏省食品药品检验所,南京210008 [2]中国药科大学药物分析教研室,南京210009

出　　处：《中国药学杂志》2014年第7期596-603,共8页Chinese Pharmaceutical Journal

摘　　要：目的采用9-芴甲氧羰酰氯(Fmoc-Cl)为柱前衍生化试剂,建立了Fmoc-Cl柱前衍生化反相高效液相色谱法用于检查并鉴定硫酸依替米星的有关物质。方法采用Lichrospher-C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.1%的甲酸溶液,等度洗脱,流速1.0 mL·min-1,检测波长265 nm,并采用电喷雾正离子化监测的LC-TOF/MS和LC-MS/MS鉴定主要有关物质。结果该法比现有文献报道方法灵敏度和准确性更好,衍生化反应完全,衍生物稳定,依替米星及其有关物质衍生物色谱分离度良好,共测得并鉴定了硫酸依替米星中的25个有关物质,其中8个为第一次报道。结论方法专属性高、准确度和耐用性好,适用于硫酸依替米星的含量测定及有关物质检测。OBJECTIVE To identify the related substances in etimicin sulfate by hyphenated techniques using pre-column deri- vatization with 9-fluorenylmethyloxycarbonyl chloride. METHODS The analysis was carried out on a Lichrospher-C18 column （4. 6 mm ×250 mm,5 ×m）by using acetonitiile-0. 1% formic acid solution as the mobile phase with isocratie elution mode at a flow rate of 1.0 mL . min-1, and the detection was performed at 265 nm. LC-TOF and LC-MS/MS methods were employed for the identification of the major related substances. RESULTS It was proved that the reaction was complete and the derivatives were stable. Good retention and separations were achieved. Twenty-five impurities were proposed based on comparison of their fragmentation patterns including 8 newly identified, and their formation mechanisms were also speculated. CONCLUSION The established method is specific and ac- curate with very good ruggedness and thus is suitable for the determination of etimicin sulfate and its related substances.

关 键 词：依替米星 庆大霉素C1a 奈替米星 9-芴甲氧羰酰氯 衍生化 

分 类 号：R917[医药卫生—药物分析学]