In 2018, a group of angiotensin II receptor blockers – sartans, were recalled from the market due to the presence of an impurity called NDMA (N-nitroso dimethylamine). Since then, there has been increased scrutiny of drug products for nitrosamines impurities. Nitrosamines are a type of impurity that belong to the carcinogens family and can form during the production of pharmaceutical products or substances when certain nitrogenous agents (amides, secondary amines, urea derivatives, carbamates, and other) react with a nitrogen atom in a +3 state.
The ICH M7(R1) Guideline classifies nitrosamines as Class 1 impurities, which are a significant concern. The presence of these impurities in pharmaceutical products can be attributed to a variety of factors, such as contaminated raw materials in API production, the use of nitrosating agents and recovered materials, nitrosamines contaminated starting materials and intermediates, and specific packaging materials and degradation processes.
In response to this issue, the EMA has requested marketing authorization holders to review drug products containing synthetized APIs for nitrosamines impurities content. At Selvita, we have the expertise and experience to support our clients throughout the testing process, from method development and cGMP validation to routine batch analysis. Our team is well-versed in working with challenging matrices and detecting impurities at trace levels, which is particularly important in the case of nitrosamines detection.
Since 2018, the detection of nitrosamines has become an important topic for the pharmaceutical industry. In August 2023, the FDA issued final guidance on Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs) . APIs that contain secondary amine or dimethyl tertiary amine centers are at risk of forming NDSRIs by nitrosation of the amine center. To follow this guidance, manufacturers of approved APIs and drug products are directed to evaluate nitrosamines risks within the next three months, and the guideline specifies that by August 2025, any NDSRIs in drug products or APIs must comply with the FDA-recommended AI limits and corresponding carcinogenic potency categories.
At Selvita, we have taken steps to provide efficient support for pharmaceutical companies by adapting our analytical equipment to accurately detect nitrosamines within the required limits and in the matrices in which they should be determined.
Over time, the focus on nitrosamine determination has expanded to include not only the seven basic nitrosamines but also those derived from active pharmaceutical ingredients (API). Given that about 80% of APIs contain nitrogen atoms in their structures, there is a wide range of possibilities for the formation of this type of nitrosamine. These nitrosamines are classified as unknown genotoxic impurities according to the ICH M7(R1) Guideline, and they are required to be analyzed with a limit of 1.5 µg/day.
At Selvita, we have the necessary experience and expertise to develop methods that meet these limits and detect these unknown genotoxic impurities. Below is a list of nitrosamines for which analytical methods have been developed and validated under full GMP/GLP requirements.
| API | Formulation | Nitrosoamines | LOQ | Technique |
|---|---|---|---|---|
| Apremilast | API | NDMA | 24 ng/mL | HS-GCMS-Q |
| Bifenox | API | NDMA | 50 ng/mL | HS-GCMS-Q |
| Bisoprolol fumarate | tablets | API derived nitrosamine | 0.45 ng/mL | LCMS-QQQ |
| Glatimer acetate | API | NDMA; NMBA; NDEA; NDBA; NMPA; NEIPA; NDIPA | 0.7 ng/mL; 10 ng/mL; 2 ng/mL; 0.5 ng/mL; 0.3 ng/mL; 0.3 ng/mL; 0.3 ng/mL | GCMS-QQQ |
| Imatinib mesylate | tablets | API derived nitrosamine | 0.45 ng/mL | LCMS-QQQ |
| Losartan potassium | API | NDEA | 20 ng/mL | GCMS-Q |
| Metformin | tablets | NDMA | 0.5 ng/mL | GCMS-QQQ |
| Metoprolol | tablets | API derived nitrosamine | 0.30 ng/mL | LCMS-QQQ |
| Niclosamide | tablets | NMBA | 50 ng/mL | LCMS-QQQ |
| Olmesartan medoxomil | API | NDMA, NDEA | 25 ng/mL; 25 ng/mL | HS-GCMS-Q |
| Prednisolone | API | NDMA, NDEA, NEIPA, NDIPA, NDBA | 0.9 ng/mL; 0.3 ng/mL; 0.3 ng/mL; 0.5 ng/mL; 0.9 ng/mL | GCMS-QQQ |
| Prednisolone | tablets | NDMA, NDEA, NEIPA, NDIPA, NDBA | 0.5 ng/mL; 0.5 ng/mL; 0.5 ng/mL; 0.5 ng/mL; 0.5 ng/mL | GCMS-QQQ |
| Ranitidine | tablets | NDMA | 1 ng/mL | LC-HRMS |
| Sunitinib | capsules | NDMA, NDEA, ENEDA | 50 ng/mL | LCMS-QQQ |
| Telmisartan | API | NDMA, NDEA | 0.5 ng/mL; 0.15 ng/mL | GCMS-QQQ |
| Telmisartan | API | NDMA, NDEA | 6 ng/mL; 12 ng/mL | HS-GCMS-Q |
| Telmisartan | API | NDBA | 10 ng/mL | GCMS-Q |
| Tramadol | capsules | NDMA, NDEA, NDBA | 2.0, 0.5, 0.5 ng/mL | LCMS-QQQ |
Our setup:
- Agilent 8890 gas chromatographs with MS 5977B detectors and optional headspace 7697A samplers/split-spitless injector/multimode injector
- Thermo Scientific TRACE 1310 gas chromatograph with MS QQQ TSQ 9000 detector and optional headspace sample/ split-spitless injector/PTV injector
- Thermo Scientific TSQ Altis mass spectrometer with Dionex UltiMate 3000 RS UHPLC system
- Thermo Scientific TSQ Altis Plus mass spectrometer with Vanquish Flex Duo LPG UHPLC system – 2 pieces
- Thermo Scientific TSQ Altis Plus mass spectrometer with Vanquish Flex HPG UHPLC system
- Agilent 6470 LC/TQ mass spectrometer with 1290 Infinity II UPLC system
- Thermo Scientific Orbitrap Exploris 120 high-resolution mass spectrometer with Vanquish Flex Duo LPG UHPLC system
At Selvita, we have a dedicated team of analytical specialists with extensive experience in the determination of nitrosamines. They are well-versed in the analytical challenges associated with this type of determination and are committed to providing an individualized approach for each request.
When working with our clients, we consider all relevant factors, such as the structure of the nitrosamine being analyzed, the required determination limits, and the nature of the matrix. Based on this information, we determine which analytical technique to use and how to prepare the test sample solution. We also strive to provide an approximate time frame for the project, which can vary from case to case but generally falls within the following ranges:
- Development of the analytical method: 2 – 4 weeks
- Validation of the method for one product with reporting: between 3 – 4 weeks
- Routine analysis or screening analysis of the sample using already existing method: 1 – 2 weeks
If you would like to learn more about our nitrosamine determination services, please do not hesitate to contact us.