Overview of nitrosamine testing requirements in the EU

Nitrosamines (more formally known as N-nitrosamines) are organic chemical compounds that form when nitrosating agents react with certain amines. The compounds do not have known commercial uses, but nitrosamine impurities may be unintentionally present in pharmaceuticals, food, cosmetics, and rubber-based materials. 

Studies have demonstrated a connection between long-term exposure to nitrosamines and increased risk of cancer, leading to several N-nitrosamines being classified as class 1B carcinogens under the EU REACH Regulation.¹ Due to these concerns, strict limits apply to nitrosamine impurities in high-risk product groups.

Nitrosamine migration from rubber articles

Rubber and elastomers may contain nitrosamine impurities originating from the nitrosation of certain rubber additives. One concern is that infants may be exposed to nitrosamines and nitrosamine precursors (nitrosatable substances) through teats and soothers manufactured using these materials. In the EU, nitrosamine release from teats and soothers must not exceed the following limits, set in Directive 93/11/EEC:²

• 0.01 mg in total of nitrosamines released/kg (of parts made of elastomer or rubber),

• 0.1 mg in total of nitrosatable substances/kg (of parts made of elastomer or rubber).

The same limits also apply to children’s drinking accessories under European standard EN 14350.

Compliance testing is performed in accordance with the EN 12868 standard. The sample material is immersed in a test-release liquid for 24 hours, after which N-nitrosamines are extracted from the solution using dichloromethane and detected using gas chromatography.

Nitrosamines in food and food contact materials

The European Food Safety Authority (EFSA) adopted a new scientific opinion on 10 carcinogenic N-nitrosamines in food in 2023, concluding that they do raise a health concern.³ Meat and meat products were found to be the main food category contributing to nitrosamine exposure, but the substances have also been detected in fish products, beer and other beverages, cheese, soy sauce, oils, processed vegetables, and human milk.

Following EFSA’s risk assessment, a European Commission working group started preparing a recommendation on monitoring N-nitrosamine presence in food, identifying factors resulting in high nitrosamine levels, and gathering information on measures that could reduce these levels.⁴ The monitoring recommendation is expected to target the ten carcinogenic nitrosamines identified in the EFSA opinion. These are listed in Table 1.

Especially in meat, fish, and cheese products, N-nitrosamine formation is closely linked to the use of nitrites and nitrates as preservatives. To mitigate the risks, the maximum levels for certain nitrates and nitrites as food additives were lowered after EFSA’s risk assessment.⁵ Limits also apply to the presence of nitrates as impurities in certain leafy vegetables and baby food.⁶

Another way in which nitrosamines end up in food is migration from rubber and elastomers used to manufacture food contact materials (FCMs), such as nitrile gloves, hoses, seals, conveyor belts, and food containers. As there is no harmonized EU-level legislation that would specify migration limits for such materials, German BfR Recommendation XXI/1 is often used as the safety reference. According to the recommendation, nitrosamine migration from elastomers to food must not exceed 1 μg/dm².⁷

Nitrosamine migration from toys

Directive 2009/48/EC on toy safety prohibits nitrosamines and nitrosatable substances in toys intended for children under three years of age and other toys intended to be placed in the mouth, such as balloons.⁸ The migration limits are 0.05 mg/kg for N-nitrosamines and 1 mg/kg for N-nitrosatable substances, and compliance testing is performed in accordance with standard EN 71-12. 

According to the European Commission’s Safety Gate rapid alert system for dangerous products, balloons are the most common toy category where products have to be withdrawn from the market due to non-compliance with nitrosamine limits.⁹ Manufacturers should have such products tested for nitrosamine impurities before placing them on the market to avoid negative financial and reputational repercussions related to product recalls.

Nitrosamines in pharmaceuticals

The European Medicines Agency (EMA) sets limits on the maximum acceptable intake (AI) of around 200 nitrosamines in human medicinal products.¹⁰ AI limits apply to both small-molecule nitrosamines, such as NDMA, NMEA, NDEA, and NPYR, which can form in various types of drug products, and nitrosamine drug substance-related impurities (NDSRIs), which form through the nitrosation of specific active pharmaceutical ingredients (APIs) and are thus unique to them.

As the AI levels are very low (in the nanograms-per-day range), highly sensitive analytical methods, such as GC-MS/MS or LC-MS/MS, are needed for compliance testing. Several small-molecule nitrosamines can be screened from pharmaceuticals simultaneously, while NDSRIs require targeted analysis. A feasibility study is typically required before routine analysis is possible, as the method will have to be validated for the specific matrix.

More information about the risk assessment process and testing options can be found in our article on nitrosamine impurity analysis of pharmaceuticals.

Nitrosamines in cosmetics

Nitrosamines are listed as prohibited substances in Annex II of Regulation (EC) No 1223/2009 on cosmetic products.¹¹ To mitigate their formation, high-risk precursor chemicals, such as many hair dye substances, must not be used with nitrosating agents and must be kept in nitrite-free containers. The nitrosamine content in the final cosmetic product must not exceed 50 μg/kg.

Our testing solutions

Measurlabs provides analysis services to help evaluate various products’ compliance with European nitrosamine regulations and guidelines. The following are some popular examples:

• Release of N-nitrosamines and N-nitrosable substances from rubber and elastomers, including teats, soothers, and food contact materials

• Routine screening of pharmaceuticals and cosmetics for common small-molecule nitrosamines

• Custom targeted analyses for API-specific nitrosamines, such as N-nitroso-fluoxetine and N-nitroso-duloxetine

For further information or a quote for your analysis project, please contact us using the form below.