目的:建立二维液质联用方法确定多黏菌素E甲磺酸钠(CMS)杂质的结构及来源,进而用于药物质量控制研究。方法:一维系统:采用Acquity UPLC® Peptide CSH C18(150 mm×2.1 mm,1.7 μm)色谱柱,以磷酸盐缓冲液(7.8 g·L-1磷酸二氢钠,用1 mol·L-1氢氧化钠溶液调节pH至6.4)-乙腈(19∶1)为流动相A,磷酸盐缓冲液-乙腈(1∶1)为流动相B,梯度洗脱,流速0.3 mL·min-1,柱温30 ℃;二维系统:采用Acquity BEH C18柱(50 mm×2.1 mm,1.7 μm)色谱柱,以甲酸铵(A)-乙腈(B)为流动相,梯度洗脱,流速0.2 mL·min-1,柱温40 ℃,检测波长210 nm。质谱检测器采用ESI源,负离子扫描模式。结果:采用2D-LC-Q TOF MS法,推定了CMS中55个杂质的结构,并推测其主要来源为多黏菌素E1-I、多黏菌素E1-7MOA、多黏菌素E3及多黏菌素E6。结论:利用二维液质联用技术推定CMS中杂质峰的结构及来源,并用来评价不同厂家、生产工艺的原料药中杂质含量变化,有利于改进生产工艺,从源头控制药物质量。
Objective: To establish a suitable method to determine the structure and source of impurities of colistimethate sodium (CMS) for drug quality control studies. Methods: Frist-dimensional system: using Acquity UPLC® Peptide CSH C18(150 mm×2.1 mm, 1.7 μm) column, the mobile phase A was phosphate buffer (7.8 g·L-1 sodium dihydrogen phosphate, adjusted to pH 6.4 with 1 mol·L-1 sodium hydroxide)- acetonitrile (19∶1), the mobile phase B was phosphate buffer-acetonitrile (1∶1). Gradient elution was performed at a flow rate of 0.3 mL·min-1. The column temperature was 30 ℃. Second-dimensional system: the Acquity BEH C18 column (50 mm×2.1 mm, 1.7 μm) column was used with ammonium formate(A)-acetonitrile mixture as mobile phase with gradient elution. The flow rate was 0.2 mL·min-1. The column temperature was 40 ℃. The detection wave length was 210 nm. The ESI source was used in negative ion mode. Results: The 2D-LC-Q TOF MS method was used to infer the structure of the 55 impurities in CMS, and the main sources were polymyxin E1-I, polymyxin E1-7MOA, polymyxin E3 and polymyxin E6. Conclusion: The structure and source of impurities in CMS are determined by 2D-LC-Q TOF MS, and the changes in the content of impurities such as manufacturers and production processes are evaluated, which is conducive to improving the production process and controlling drug quality at the source.
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