目的: 采用检测厄贝沙坦中N-亚硝基二乙胺（NDEA）的高分辨LC-Q TOF MS法对未知峰[相对保留时间（RRT）为0.95]进行调查,确定该未知峰结构及产生来源,制定出相应的分析方法优化策略。方法: 采用LC-Q TOF MS全扫描的方法对该未知峰进行质谱全扫描,确定出未知峰的结构式,比较该未知峰与NDEA一级质谱及二级质谱的差异,并参照LC-MS/MS方法对待测组分的定量离子选择,阐述NDEA检测中该未知峰出现的原因,制定相应的分析方法优化策略。结果: 根据LC-Q TOF MS分析结果,推测该未知峰为1-戊酰胺,这是沙坦原料药样品中含有的微量常规杂质,并通过与1-戊酰胺对照品比对进行最终确认。由于1-戊酰 胺摩尔相对分子质量比NDEA少1,因此,在ESI正离子模式下,1-戊酰胺氢离子加合离子[M+H]++1的同位素峰m/z与NDEA [M+H]+离子m/z相同,即103,并且与NDEA类似,1-戊酰胺氢离子加合离子[M+H]++1同位素峰在MRM模式下也能裂解1个乙烯中性分子,产生m/z 75同位素离子,导致当选择m/z 103作为母离子,m/z 75 为定量子离子时,1-戊酰胺会对NDEA的检测产生干扰。结论: 在沙坦类药物NDEA LC-MS/MS检测中RRT 0.95的未知峰为1-戊酰胺,是沙坦类药物生产过程中产生的常规微量工艺杂质,而不是另外的亚硝胺杂质;在后续的研究中,将采用特征选择性更强的m/z 47 定量子离子模式,可以避免该未知峰产生。
蔡虹
,
谢鹤
,
李晓玲
,
赵诗怡
,
荆冰江
,
金建阳
,
陈文斌
,
叶坚
,
李敏
,
林金生
. 基于LC-MS/MS法对N-亚硝基二乙胺检测中的未知峰调查及排除策略[J]. 药物分析杂志, 2025
, 45(2)
: 265
-274
.
DOI: 10.16155/j.0254-1793.2024-0383
Objective: To investigate an unknown peak at relative retention time (RRT) 0.95 of N-nitrosodiethylamine (NDEA) of irbesartan API with the LC-MS/MS detection method. To elucidate the structure and origin of this unknown peak. Its structure was confirmed with a reference compound, and a method optimization strategy was proposed to eliminate the impact of this unknown peak. Methods: The unknown peak causing interference in the determination of NDEA within sartan drug substances was examined using a full scan method of LC-MS/MS analysis. The unknown peak at RRT 0.95 was validated by conducting high resolution LC-MS analyses and comparing with that of a reference sample. Additionally, a strategy for resolving this issue was proposed in the further investigation. Results: According to the LC-Q TOF MS results, the unknown peak at RRT 0.95 was confirmed to be 1-pentanamide, a trace regular impurity commonly existing in the sartan samples. As the extract mass of 1-pentanamide was 1 less than that of NDEA, the m/z value of an isotope in 1-pentanamide ([M+H]++1) was 103 and was equal to the m/z value of the main [M+H]+ isotope of NDEA under the MRM mode. Furthermore, the [M+H]++1 isotope ion of 1-pentanamide could occur a neutral lose by leaving a molecule of ethylene to generate the isotope ion of m/z 75, which was similar to NDEA in the MRM analysis under ESI positive mode. Therefore, when m/z 103 was used as the precursor ion and m/z 75 was used as the quantitative ion, 1-pentanamide will interfere with detection of NDEA. Conclusion: In the LC-MS/MS analysis for NDEA, the unknown peak observed at a retention time of 0.95 has been identified as 1-pentanamide. This compound is confirmed as a common trace impurity that regularly occurs during the production of sartan active pharmaceutical ingredients (APIs), rather than being another nitrosamine impurity. Subsequent research has revealed that employing a more selective quantitative model and choosing m/z 47 as the quantifier ion effectively avoid the unknown peak caused by 1-pentanamide.
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