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, 5 μm),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。
Abstract
OBJECTIVE To identify the related substances in etimicin sulfate by hyphenated techniques using pre-column derivatization with 9-fluorenylmethyloxycarbonyl chloride. METHODS The analysis was carried out on a Lichrospher-C18 column (4.6 mm×250 mm,5 μm)by using acetonitrile-0.1% formic acid solution as the mobile phase with isocratic 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 accurate 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
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Key words
etimicin /
gentamicic C1a /
netilmicin /
9-fluorenylmethyloxycarbonyl chloride /
derivatization
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中图分类号:
R917
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参考文献
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脚注
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