LC-DAD-SPE-NMR/MS技术用于卡巴他赛注射液中微量未知杂质的鉴定
收稿日期: 2024-07-09
网络出版日期: 2024-11-13
基金资助
国家自然科学基金青年项目(31400304);波谱与原子分子物理国家重点实验室开放基金(T151203);中国科学院仪器功能开发项目(YG2011095);中国科学院仪器功能开发项目(YG2012110);中国科学院仪器功能开发项目(YG2012108)
Identification and Structural Characterization of an Unknown Trace Degradation Impurity in Cabazitaxel Injection by LC-DAD-SPE-NMR/MS
Received date: 2024-07-09
Online published: 2024-11-13
本研究采用液相色谱-二极管阵列检测器-固相萃取-核磁共振/质谱（LC-DAD-SPE-NMR/MS）联用技术，对卡巴他赛注射液中一个含量约为0.3%~0.4%的未知降解杂质完成化学结构解析. 卡巴他赛注射液中高含量的辅料吐温-80与该杂质在液相色谱分析中不能完全分离，直接制备高纯度杂质难度较大，因此难以采用先分离-后鉴定的传统策略鉴定该杂质. 本研究采用自动化的LC-DAD-SPE-NMR/MS联用技术，通过对萃取物溶液进行NMR谱详细分析，提取出降解杂质中氢-氢、氢-碳原子间相关信息，得到原子化学位移及连接关系，成功排除吐温-80谱峰的干扰，最终鉴定降解杂质为卡巴他赛中紫杉烷母核重排后的产物. 整个实验在48 h之内完成，并以混合物分析思路完成降解杂质结构解析. 本研究是LC-DAD-SPE-NMR联用技术在微量药物杂质分析领域的一次成功应用实例，为新药研究中杂质结构解析提供了新思路.
关键词： 卡巴他赛; 降解杂质; 液相色谱-二极管阵列检测器-固相萃取-核磁共振/质谱联用技术; 结构解析; 分析方法
刘红兵 , 刘惠丽 , 罗立廷 , 孙丽娟 , 陈雷 . LC-DAD-SPE-NMR/MS技术用于卡巴他赛注射液中微量未知杂质的鉴定[J]. 波谱学杂志, 2025 , 42(1) : 34 -46 . DOI: 10.11938/cjmr20243122
In this work, the hyphenated liquid chromatography-diode array detector-solid phase extraction-nuclear magnetic resonance/mass spectrometry (LC-DAD-SPE-NMR/MS) technique is employed to characterize a novel degradation impurity with an approximate content of 0.3%~0.4% in cabazitaxel injection. In LC analysis, the impurity is co-eluted with tween-80, an important excipient contained in cabazitaxel injection, which poses a great challenge for high-purity degradation impurity isolation and further structure elucidation. Based on LC-DAD-SPE-NMR/MS hyphenation, automatic SPE is performed to trap and concentrate the impurity, and extensive NMR analysis was performed to characterize the structure of the impurity. In the 1H NMR spectrum, partial peaks of the impurity are overlaid with co-eluted tween-80. To navigate the complexity, 1H-1H, 1H-13C correlations, 1H and 13C chemical shifts are extracted from the interfering peaks of tween-80 based on extensive analysis of 2D NMR spectra. Finally, the structure of the impurity is elucidated to be a cabazitaxel isomer with a rearranged taxane skeleton. The elucidation is completed in less than 48 h, showcasing a successful application of LC-DAD-SPE-NMR/MS hyphenation in determining the structure of an unknown trace impurity in pharmaceutical drugs, which evidences its capability and application prospects in the pharmaceutical industry.
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