Nitrosamine Test in Drugs: Detection of Carcinogenic Contaminants and Nano Test Solutions
The pharmaceutical industry experienced a major crisis in 2018 due to N-nitrosodimethylamine (NDMA) contamination found in blood pressure medications from the “Sartan” group. This incident led to the recall of millions of drugs worldwide and prompted regulators to fundamentally change their approach to Nitrosamine Tests on Drugs. Today, the control of nitrosamine contamination has become an integral part of drug development and production processes.
Nitrosamines are compounds whose chemical structure contains an n-nitroso group (R1N(-R2)-N=O). Many of them have shown a strong carcinogenic (carcinogenic) effect in animal experiments. Their formation in drugs can depend on numerous factors, including the synthesis of raw materials, the manufacturing process, packaging materials and even the storage conditions. Therefore, continuous monitoring using precise and reliable analytical methods is essential in order not to expose patients to these harmful compounds.
Sources and Mechanisms of Nitrosamine Contamination
Nitrosamines can get into drugs in various ways or spontaneously form in them. The most common mechanism of formation is the reaction of secondary or tertiary amines with nitric acid (HNO₂) or nitrite salts. This reaction is accelerated under acidic conditions.
- Comes from raw materials: Solvents, catalysts or starting materials used in the synthesis of the pharmaceutical agent (API) may contain nitrosamine impurities.
- Process-related: Certain excipients (excipien) used during production may contain nitrite and react with the amines in the API.
- Due to packaging: In particular, blister packaging containing nitrocellulose-based inks may cause the migration of volatile nitrosamines into the drug.
- Storage conditions: High humidity and temperature are factors that accelerate the formation of nitrosamines.
Limits of the supervisory authorities and current legislation
The European Medicines Agency (EMA), the US Food and Drug Administration (FDA) and the Turkish Drugs and Medical Devices (TİTCK) have set extremely low limits for nitrosamine contaminants. These limits were calculated based on a “lifetime exposure scenario” and aim to keep the risk of cancer at a negligible level.
For NDMA, one of the most common nitrosamines, the daily allowable intake (AI) was set at 96 ng/day. This means that the maximum amount of NDMA that may be included in the daily dose of a drug is one part per billion (PPB). In order to be able to detect such low values, highly sensitive analysis techniques are required.
The regulatory authorities require the holders of a market license (admission holders) to carry out a risk assessment for all their products and to carry out confirmation examinations for products where a risk has been identified. This process is done in three steps: "Step 1: Risk Assessment", "Step 2: Confirming Tests" and "Step 3: Change Management".
Frequently monitored nitrosamine impurities
- NDMA (N-Nitrosodimethylamine): One of the most common and strongest carcinogenic nitrosamines.
- DEA (N-N-Nitrosodiethylamine): also detected in sartan group drugs and some antihistamines.
- NMBA (N-nitroso-N-methyl-4-aminobutyric acid): was detected in drugs such as valsartan.
- Nipea (N-nitrosoethyl isopropylamine): was found in some generics.
- NDSRI (nitrosamine drug-related contaminants): These are new generation nitrosamines that come from the active ingredient itself and have been the focus in recent years.
Common Nitrosamines and Limits for Daily Daily Reception (AI)
The following table summarizes the nitrosamine contaminants that are most frequently detected in drugs and the limit values for the permissible daily intake set by the supervisory authorities:
| Name of nitrosamine | Chemical formula | Daily Allowed Limit (AI) | Carcinogenicity class |
|---|---|---|---|
| NDMA (N-nitrosodimethylamine) | C₂H₆N₂O | 96 ng/day | IARC Group 2A (probably carcinogenic) |
| DEA (N-nitrosodiethylamine) | C₄H₁₀N₂O | 26.5 ng/day | IARC Group 2A (probably carcinogenic) |
| NMBA (N-nitroso-N-methyl-4-aminobutyric acid) | C₅H₁₀N₂O₃ | 96 ng/day | IARC Group 2B (possibly carcinogenic) |
| Nipea (N-nitrosoethyl isopropylamine) | C₅H₁₂N₂O | 26.5 ng/day | IARC Group 2B (possibly carcinogenic) |
Analytical Methods for Nitrosamine Analysis
The detection of nitrosamines requires extremely sensitive analytical techniques due to the low detection limits in the PPB range (parts per billion). Conventional HPLC methods often cannot guarantee this sensitivity.
1. LC-MS/MS (Liquid Chromatography - Mass Spectrometry)
Currently, this is the gold standard for nitrosamine analysis. Thanks to their high selectivity and sensitivity, nitrosamines can be reliably detected even in complex drug matrices at very low concentrations. Multi-response monitoring mode (MRM) minimizes false-positive results by tracking specific ion transitions.
2. GC-MS/MS (Gas Chromatography - Mass Spectrometry)
Suitable for volatile nitrosamines (like NDMA, NDEA). Can be used with headspace or direct injection techniques. However, the method may not be suitable for some thermally unstable nitrosamines.
3. HRMS (High Resolution Mass Spectrometry)
A powerful tool for identifying and structurally elucidating unknown nitrosamine contaminants. plays a crucial role, especially in the discovery of NDSRI.
GLP compliant nitrosamine test services with nano test
Conducting nitrosamine risk assessments and verification tests for drug manufacturers requires extensive expertise and appropriate infrastructure.
Nano-Test is the most reliable solution partner in the pharmaceutical industry with its GLP-compliant analysis services in this area.
Description of Nano-Test Services
Nano-Test offers the pharmaceutical industry comprehensive services in the field of Nitrosamine Contaminant Analysis. Our labs accredited according to GLP (Good Laboratory Practice) and ISO/IEC 17025 are analyzed that correspond to all current limits set by the EMA, FDA and TİTCK.
Our scope of services includes the detection and quantitative determination of all common nitrosamines, including NDMA, NDEA, NMBA, NIPEA and NDSRI. In our analyzes, the latest devices such as LC-MS/MS, GC-MS/MS and HRMS are used. Fully validated methods are applied for each analysis, which correspond to the corresponding pharmacopoeia monographs and the current guidelines of the supervisory authorities.
As a nano test, we not only deliver a test result, but also create complete and well-founded GLP compliance reports that you can use for your approval applications and that are approved for inspections. We accompany you in all processes, from risk assessment to the confirmation examinations.
The advantages of nano-test in nitrosamine analysis:
- Extremely low detection limits: Detection performance below the PPB range (ng/g) using LC-MS/MS.
- Complete validation: Method validation according to ICH Q2(R1) and the relevant guidelines.
- Fast reporting: Result delivery within 10 working days for standard analyses.
- Specialist team: A team of chemists specializing in mass spectrometry and contamination profiles.
- Current law tracking: Immediate adjustment to the latest changes to the limit values published by the EMA and the FDA.
Nitrosamine contaminants are among the most serious quality and safety issues in the pharmaceutical industry. In order to ensure patient safety and meet the requirements of the supervisory authorities, the Nitrosamine Test for Drugs is no longer an option, but an obligation. Choosing the right analytical methods, complete validation and reporting results in accordance with GLP standards are critical.
Nano-Test supports drug manufacturers with its state-of-the-art technological infrastructure and its expert team in managing nitrosamine risks. Have your nitrosamine analyzes performed with Nano-Test's warranty to ensure smooth access of your products to global markets.