The nitrosamine testing landscape
31 May 2026
Nitrosamines are chemical compounds that form when nitrites react with amines. They are found in food and beverages but also in drugs and cosmetics and in tobacco products as tobacco-specific nitrosamines (TSNAs). Dean Hatt outlines the importance of testing for them across relevant sectors...
Nitrosamines require certain conditions such as heat, pH or processing to form. Nitrites can be found in foods, preservatives, the body and soil, among other areas, while amines are organic compounds derived from ammonia, where one or more hydrogen atoms are replaced by carbon-containing groups, occurring naturally.
Following an increased focus from regulators including the Food and Drug Administration (FDA) and European Medicines Agency (EMA), testing has since ramped up, uncovering nitrosamines in pharmaceutical products including sartans, metformin and ranitidine.
Nitrosamines form through a nitrosation reaction when amines, particularly secondary amines, react with nitrosating agents such as nitrite under acidic conditions (e.g., in the stomach), with heat, processing, and storage further promoting their formation. While nitrosamines themselves are not highly reactive, they are potent procarcinogens that become harmful after metabolic activation by liver enzymes (e.g., CYP450), generating reactive intermediates such as alkylating (diazonium) species. These intermediates can damage DNA by forming adducts like O?-methylguanine, leading to mutations and increased cancer risk.
The most notorious are N-Nitrosodimethylamine (NDMA) and N-Nitrosodiethylamine (NDEA), which are found in processed food, mostly meat and fish, and as a drug contaminant. NNK (4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone) is a highly potent tobacco-specific nitrosamine (TSNA) that is produced through the nitrosation of nicotine, occurring naturally in tobacco leaves, as is Nitrosonornicotine (NNN) which is produced during the curing and processing of tobacco.
Nitrosamine testing
A full suite of Harmful and Potentially Harmful Constituents (HPHC) testing provides a comprehensive approach to identifying and quantifying toxic compounds in tobacco and nicotine products in line with guidance from regulators. Techniques like ICP-MS, LC-MS and CG-MS are used here to ensure complete product characterisation.
Typically, this includes analysis of key chemical classes such as carbonyl compounds (e.g. formaldehyde, acrolein), volatile organic compounds (benzene and 1,3-butadiene) along with TSNAs and toxic metals ensuring a comprehensive understanding of product emissions and exposure risk.
Partnering with a specialist testing organisation enables product developers to accurately detect and quantify nitrosamines and other harmful compounds at trace levels, supporting product safety evaluations, risk assessments and regulatory submissions.
Regulation
Specific regulations of nitrosamines are based on the type of the product, accounting for their risks in the broader chemical safety framework. While there are no universal limits on nitrosamines, regulation focuses on precursors. For example, in the UK and EU, nitrites are limited to less than 150mg/kg in most cured meats, whereas 150-300mg/kg is allowed in long-cured products to help guard against any subsequent health risks. Drinking water has similar standards while TNSAs have maximum levels set that must be closely analysed.
Since 2018, products have been recalled in the pharmaceutical market when the focus on nitrosamine testing intensified after featuring in antibiotics and treatments of other common conditions including heartburn and diabetes. Across regulators in Europe (EMA), the United States (FDA) and the UK’s Medicines and Healthcare Regulatory products Agency (MHRA), risk assessments of nitrosamine formation typically have low acceptable intake limits, often as little as nanograms per day, based on lifetime cancer risk.
Cosmetics are regulated under general chemical safety legislation where nitrosamines are prohibited or must be present at very low levels. Some ingredients are formaldehyde-releasing preservatives which can become nitrosating agents where manufacturers must avoid conditions that promote formation.
Without adequate nitrosamine testing capabilities and processes, manufacturers across many industries risk delay to regulatory approval, reformulation of products and reputational damage.
Pic: Broughton laboratory
Dean Hatt is senior consultant toxicology at testing and regulatory consultant Broughton
