NNK遗传毒性杂质及其异构体检测方法的探索性研究
Exploratory Study on Detection Methods of NNK Genotoxic Impurity and Its Isomers
关皓月 1刘年 1许凯 1牛剑钊 1杨锐1
作者信息
- 1. 中国食品药品检定研究院,北京 100050
- 折叠
摘要
亚硝胺类化合物由于其结构上存在N-N=O键,容易发生异构化现象.本研究在采用HPLC法对4-(甲基亚硝胺基)-1-(3-吡啶基)-1-丁酮(NNK)进行分析时,观察到双色谱峰现象.并采用PFP色谱柱对NNK及其异构体进行有效分离,采用LC/MS二级质谱扫描确认了色谱图中2个色谱峰的分子、离子以及碎片离子均相同.同时采用NMR法对一级和二级核磁信号进行结构归属,进一步确证NNK中的杂质峰均属于同分异构体.此外,通过变温试验,采用NMR法测定了NNK的异构化比例范围;通过HPLC的变温试验,确证了 NMR的结果.本研究通过多角度试验方法验证NNK存在互变异构体的现象,有利于NNK含量的准确分析,同时为该类异构化遗传毒性杂质的检测提供了理论依据.
Abstract
Nitrosamine compounds,due to the N-N=O bonds in their structure,can easily cause isomerization.When an HPLC method was used to analyze 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK),double peaks were observed.NNK and its isomers were effectively separated by PFP chromatographic column.Then a LC/MS method was used to investigate the secondary fragment ions of the two separated sample peaks.NMR investigation further confirmed that the impurity peaks in NNK belonged to its isomer,and the isomerization ratio range of NNK was determined by NMR quantitative method through variable temperature experiment.And the NMR results were confirmed through the temperature change test of HPLC.This study verified structure of the isomer through a variety of methods,which was conducive to the accurate analysis of NNK content and provided a theoretical basis for the detection of genotoxic impurities.
关键词
4-(甲基亚硝胺基)-1-(3-吡啶基)-1-丁酮(NNK)/异构体/核磁定量/LC-MS/变温试验Key words
4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)/isomer/nuclear magnetic quantification/LC-MS/variable temperature experiment引用本文复制引用
基金项目
国家"重大新药创制"科技重大专项基金资助项目(2017ZX09101001)
出版年
2023
NETL
NSTL
万方数据