Danis Ibrahim, Unal Durisehvar Ozer
Department of Analytical Chemistry, Faculty of Pharmacy; Drug Research Center; Institute of Health Science, Istanbul University, Istanbul, Turkey.
Department of Analytical Chemistry, Faculty of Pharmacy; Drug Research Center, Istanbul University, Istanbul, Turkey.
Eur J Mass Spectrom (Chichester). 2025 Feb;31(1-2):51-62. doi: 10.1177/14690667251328823. Epub 2025 Mar 21.
In 2018, the Food and Drug Administration and the European Medicines Agency identified nitrosamine impurities in certain drugs, prompting detailed investigations by international regulatory authorities. According to ICH M7 (R1) guidelines, nitrosamines are classified as Class 1 substances, the most hazardous category, based on their carcinogenic and mutagenic properties. They are also recognized as probable human carcinogens by the International Agency for Research on Cancer. Since nitrosamine-induced DNA damage poses significant health risks, identifying potential nitrosamines in pharmaceutical products is crucial. In this study, a simple and efficient extraction method was designed to minimize matrix effects. These effects were evaluated using calibration curves prepared for each drug product in their respective matrices. The developed method was performed using an Agilent 1260 series HPLC system and an Agilent 6460 triple quadrupole tandem mass spectrometer. An Inertsil ODS-3 C18 (5 µm, 4.6 × 150 mm) column was employed for chromatographic separation. A triple quadrupole mass detector with electrospray ionization was used for detection, and multiple reaction monitoring was employed for quantification. The correlation coefficients (r²) were at least 0.999 for all eight nitrosamines. Limit of detection and limit of quantification values were determined as 0.05-0.8 ng/mL and 0.1-2.0 ng/mL, respectively. Validation results demonstrated satisfactory selectivity, accuracy, precision, and stability. The results demonstrated that the developed method is capable of reliably detecting potential nitrosamines present in pharmaceutical products at trace levels. This method contributes significantly to pharmaceutical safety and can serve as a valuable tool for future analyses.
2018年，美国食品药品监督管理局和欧洲药品管理局在某些药物中发现了亚硝胺杂质，促使国际监管机构展开详细调查。根据国际人用药品注册技术协调会M7（R1）指南，亚硝胺因其致癌和致突变特性被归类为1类物质，即最危险的类别。国际癌症研究机构也将其认定为可能的人类致癌物。由于亚硝胺诱导的DNA损伤会带来重大健康风险，因此识别药品中的潜在亚硝胺至关重要。在本研究中，设计了一种简单高效的提取方法，以尽量减少基质效应。使用在各自基质中为每种药品制备的校准曲线对这些效应进行评估。所开发的方法使用安捷伦1260系列高效液相色谱系统和安捷伦6460三重四极杆串联质谱仪进行。采用Inertsil ODS - 3 C18（5 µm，4.6×150 mm）色谱柱进行色谱分离。使用带有电喷雾电离的三重四极杆质量检测器进行检测，并采用多反应监测进行定量。所有八种亚硝胺的相关系数（r²）均至少为0.999。检测限和定量限分别确定为0.05 - 0.8 ng/mL和0.1 - 2.0 ng/mL。验证结果表明选择性、准确性、精密度和稳定性均令人满意。结果表明，所开发的方法能够可靠地检测药品中痕量水平的潜在亚硝胺。该方法对药品安全有重大贡献，可作为未来分析的宝贵工具。
