2D-LC-IT-TOF-MS鉴定他克莫司原料有关物质

Identification of Related Substances in Tacrolimus by 2D-LC-IT-TOF-MS

关键词:  他克莫司  在线脱盐-飞行时间-液质联用  杂质鉴定

Key words:  tacrolimus  2D-LC-IT-TOF-MS  impurity identification

摘要:

目的 应用在线脱盐-飞行时间-液质联用法，对他克莫司原料中检出的一杂质峰进行结构定性。方法 采用Thermo Hypersil GOLD C₁₈色谱柱（4.6 mm×150 mm，3 μm），乙腈-叔丁基甲醚-磷酸溶液为流动相，梯度洗脱；然后采用InertSustain C₁₈色谱柱（2.1 mm×50 mm，2 μm），0.1%甲酸溶液-乙腈（30∶70）为流动相，等度洗脱；电喷雾电离离子源，正离子检测模式，雾化气流速度1.5 L·min^-1，干燥气流速度10 L·min^-1，脱溶剂管温度200℃。根据多级图谱信息推定特定杂质峰结构。结果 推定出他克莫司原料中特定保留时间处色谱峰的物质结构，该检出峰不是美国药典给出的子囊霉素杂质而是有药理活性的他克莫司异构体Ⅰ，应参与含量计算。结论 建立的方法可用于他克莫司原料杂质的鉴定，为更加精准控制产品质量和工艺优化研究提供可靠的技术保证。

Abstract:

OBJECTIVE To rapidly identify the impurity peak detected in tacrolimus by 2D-LC-IT-TOF-MS. METHODS Firstly, the chromatography was Thermo Hypersil GOLD C₁₈(4.6 mm×150 mm, 3 μm) column and the mobile phase containing a mixture of acetonitrile-1,1-dimethylethyl-phosphoric acid. Then a InertSustain C₁₈(2.1 mm×50 mm, 2 μm) column was used with 0.1% formic acid-acetonitrile(30:70) as mobile phase in a gradient elution mode. Electrospray ionization source was tested in positive ion modes, nebulized gas flow was 1.5 L·min^-1. Dry gas flow was 10 L·min^-1. The desolvation tube temperature was kept at 200℃. Relatedstances were characterized according to multi-level MS behaviors. RESULTS The material structure of chromatographic peak at specific retention time in tacrolimus raw material was deduced, and the detected peak was not ascomycin impurity provided by USP but was tacrolimus isomer Ⅰ with pharmacological activity, content should be involved in calculation. CONCLUSION The established method is suitable for rapid identification of impurities in tacrolimus, which can be applied as a useful analytical tool for quality control and drug process optimization.