应用HPLC-ESI-IT-MSⁿ法推定硫酸西索米星中有关物质的结构

目的: 建立HPLC-ESI-IT-MSⁿ方法,研究硫酸西索米星的杂质谱。 方法: 采用Agilent 1100 LC /MSD Trap液质联用仪,采用Gemini NX C₁₈(4.6 mm×150 mm,5 μm)色谱柱,以水-氨水-冰醋酸(96∶ 3.6∶ 0.4) 为流动相A,甲醇为流动相B,梯度洗脱,流速1 mL·min^-1;质谱检测器采用ESI离子源,正离子检测,离子源温度为350 ℃,雾化室压力为275.8 kPa,干燥气流速为9 L·min^-1;检出杂质结构的鉴定方法根据有无杂质对照品而异:对于有对照品的杂质,通过比较其与对照品的色谱和质谱行为来确定结构;对于无对照品的未知杂质,则以已知结构物质如西索米星、奈替米星等为模型,通过分析已知物与未知杂质多级质谱行为的异同性推定其结构。 结果: 硫酸西索米星原料中检出16个有关物质,其中14个物质的结构得到归属。 结论: 本文建立的HPLC-ESI-IT-MSⁿ方法及研究结果对硫酸西索米星的质量控制和工艺评价具有参考意义。

Objective: To establish an HPLC-ESI-IT-MSⁿ method for the study of impurity profile of sisomicin sulfate. Methods: The analysis was performed by an LC-Ion Trap MSⁿ system of Agilent 1100 LC /MSD,using a Gemini NX C₁₈(4.6 mm×150 mm,5 μm) column as stationery phase,a mixture solution of water-ammonia-glacial acetic acid(96∶ 3.6∶ 0.4) as mobile phase A and methanol as mobile phase B,the gradient elution was done with a flow rate of 1.0 mL·min^-1.Electrospray ionization(ESI) source was used at positive ion detection model,the temperature of ESI source was 350 ℃,nebulizing pressure was 275.8 kPa,dry gas flow was 9 L·min^-1.The impurities which their reference substances were available were identified by comparing the chromatographic and mass spectrometric behaviors with that of reference substances,the others which reference substances were unavailable were deduced using the collision-induced dissociation(CID) spectra of sisomicin and netilmicin as templates. Results: A total of 16 compounds were characterized in commercial samples,among which 14 impurities were deduced. Conclusions: This work provides very useful information for quality control of sisomicin sulfate and evaluation of its fermentation and purification process.