Safety Monitoring

Determination of N-nitrosamine genotoxic impurities in metformin preparations by GC-MS/MS and investigation on generation mechanism of N-nitrosodimethylamine

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  • 1. Shandong Institute for Food and Drug Control, NMPA Key Laboratory for Research and Evaluation of Generic Drugs, Jinan 250101,China;
    2. Key Laboratory of Chemical Billogy of the Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan 250101,China

Received date: 2021-06-29

  Online published: 2024-06-24

Abstract

Objective: To establish a gas chromatography-tandem mass spectrometry (GC-MS/MS) method for simultaneous determination of six potential N-nitrosamine genotoxic impurities (NGIs)including N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine (NDEA), N-nitrosoethylisopropylamine (NEiPA), N-nitrosodiisopropylamine (NDiPA), N-nitrosodiphenylamine (NDPA), and N-nitrosodibutylamine (NDBA) in different metformin preparations. The generation mechanism of NDMA in metformin sustained preparations was also investigated. Methods: Chromatographic separation was achieved on capillary column using polyethylene glycol as stationary mobile phase (30 m × 0.25 mm × 0.25 μm); the column temperature was initially set at 40 ℃(0.5 min hold), and rose to 200 ℃ at a rate of 20 ℃·min-1, then ramped at a rate of 60 ℃·min-1 to 240 ℃ (5 min hold);the injection temperature was set at 250 ℃, helium and argon was used as carrier gas and collisional gas respectively; the flow rate was 1 mL·min-1and the injection volume was 2 μL. Detection was performed on tandem mass spectrometry equipped with electron impact ion source. Quantification was performed on the selected reaction monitoring (SRM) mode. Results: The six NGIs showed good linear relationships within the range of 0.25-50 ng·mL-1 (r>0.999); the detection limits were 0.01-0.07 ng·mL-1 and the quantification limits were 0.04-0.2 ng·mL-1. The average recovery fall between 97.4%and 116.0%(n= 9), and the precision, repeatability, and stability were fine with RSD value less than 10%. Only NDMA was detected in 139 batches of metformin preparations, and all samples with NDMA content exceeding the limit were metformin sustained release tablets. Results indicated that the residual dimethylamine in active pharmaceutical ingredients(API) and nitrile in excipient are the main factors to generate NDMA. Furthermore, reactive oxygen species produced by oxidation of excipients can oxidize dimethylamine in API to form NDMA. Conclusions: This method showes satisfactory sensitivity, specificity, accuracy and repeatability, and is suitable for quantification of NGIs in metformin preparations. In addition, during the preparation process of metformin sustained release tablets, the quality of API and excipients should be strictly controlled to avoid or reduce the generation of NDMA.

Cite this article

WEN Song-song, NIU Chong, LIU Qi, ZHANG Lei, GUO Chang-chuan, XU Yu-wen, WANG Wei-jian . Determination of N-nitrosamine genotoxic impurities in metformin preparations by GC-MS/MS and investigation on generation mechanism of N-nitrosodimethylamine[J]. Chinese Journal of Pharmaceutical Analysis, 2022 , 42(8) : 1424 -1432 . DOI: 10.16155/j.0254-1793.2022.08.16

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