Your Nitrosamine Impurity Testing FAQ: Answers for Regulatory Readiness and Confirmatory Analysis

Nitrosamine impurity testing is the analytical process of detecting and quantifying nitrosamines—carcinogenic compounds often formed as by-products in drug synthesis or storage. These impurities can originate from amines, nitrites, or specific reaction conditions involving solvents and catalysts. Testing is essential to comply with international guidelines and protect patient safety.
At ResolveMass, our nitrosamine impurity testing workflows involve LC-MS/MS, GC-MS, and high-resolution mass spectrometry to detect trace levels (ng/g or ppb range) of known and unknown nitrosamines.
Learn more about our analytical workflow

Some of the most frequently encountered nitrosamines include:
NDMA (N-Nitrosodimethylamine)
NDEA (N-Nitrosodiethylamine)
NMBA (N-Nitroso-N-methyl-4-aminobutyric acid)
DIPNA (N-Nitroso-diisopropylamine)
EIPNA (N-Nitroso-N-ethylisopropylamine)
NDIPA (N-Nitrosodiisopropylamine)
NDBA (N-Nitrosodibutylamine)
ResolveMass routinely performs nitrosamine impurity testing for all listed compounds and provides custom method development for client-specific targets.

The following instrumentation is typically employed:
LC-MS/MS for polar and thermally labile nitrosamines
GC-MS for volatile nitrosamines
High-resolution MS for unknown impurity screening
Our Nitrosamine Analysis Services combine these techniques with risk-based workflows and orthogonal confirmation for regulatory accuracy.

Regulatory agencies have established AI (acceptable intake) levels, usually expressed in nanograms per day:
NDMA: 96 ng/day
NDEA: 26.5 ng/day
NMBA: 96 ng/day
DIPNA & EIPNA: 26.5 ng/day
These limits form the basis of specification setting during nitrosamine impurity testing. If an impurity exceeds the AI, it must be either justified, mitigated, or the product must be reformulated or withdrawn.

Common root causes include:
Use of nitrite-containing excipients
Secondary or tertiary amines in APIs or reagents
Acidic pH environments
Recycled solvents or catalysts
Manufacturing equipment residues
Our Nitrosamine Risk Assessment Services include route analysis and synthetic process reviews to identify and mitigate these risks.

Confirmatory nitrosamine impurity testing refers to the validated, sensitive reanalysis of a sample following an initial detection. It ensures result reproducibility and supports regulatory submissions.
ResolveMass offers:
Validation per ICH Q2(R1) and FDA guidelines
Isotope dilution and internal standards
LOD/LOQ determination
Explore our confirmatory testing methods

With techniques like LC-MS/MS operating at detection limits below 10 ng/g, nitrosamine impurity testing at ResolveMass is both precise and reproducible. We perform linearity, accuracy, and robustness checks to ensure method reliability.

Yes. Using HRMS (High-Resolution Mass Spectrometry), our experts can identify and characterize unknown or novel nitrosamines—critical for regulatory response in the case of non-targeted impurity findings.
Learn about advanced impurity profiling

Typical timelines are:
Risk Assessment: 5–7 business days
Confirmatory Testing: 7–10 business days
Custom NDSRI Synthesis: 3–6 weeks
Visit our nitrosamine testing service page for expedited options.

Yes. For investigational or confirmatory work, we provide custom synthesis of Nitrosamine Drug Substance-Related Impurities (NDSRIs).
Read more about our NDSRI synthesis

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Nitrosamine impurity testing typically involves advanced analytical methods such as LC-MS/MS and GC-MS. These techniques allow for sensitive detection down to the parts-per-billion (ppb) or nanogram per gram levels. At ResolveMass Laboratories Inc., we use validated methods with isotope dilution, matrix-matched calibration, and internal standards to ensure accuracy and reproducibility across all test runs. This ensures regulatory-compliant quantification of nitrosamines such as NDMA, NDEA, and other NDSRIs.

Calculating nitrosamine content involves comparing the peak area (or intensity) of a sample against that of a known standard curve generated using authentic nitrosamine standards. The response factor, sample weight, and dilution are factored into a formula:
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Nitrosamine Content (ng/g) = (Peak Area_sample / Slope_standard curve) × Dilution Factor / Sample Weight
ResolveMass provides precise calculation of impurity concentrations as part of our standard nitrosamine impurity testing reports.

Nitrosamines are a class of genotoxic and potentially carcinogenic compounds that may form in drug products through several pathways, including:
Secondary amine reactions with nitrites under acidic conditions
Residual solvents or catalysts during synthesis
Cross-contamination from excipients or equipment
Understanding the basics of nitrosamine formation is critical for effective nitrosamine impurity testing and risk assessment. This includes knowledge of structure-reactivity relationships and potential synthetic routes of impurity formation.

Detecting nitrosamines requires highly sensitive and selective analytical techniques due to their low permissible limits and structural diversity. The most widely used methods in nitrosamine impurity testing include:
LC-MS/MS (Liquid Chromatography–Tandem Mass Spectrometry): Ideal for polar and thermally unstable nitrosamines like NMBA. Allows accurate quantification in the sub-ng/g range.
GC-MS (Gas Chromatography–Mass Spectrometry): Preferred for volatile nitrosamines such as NDMA and NDEA. Suitable for detecting low molecular weight impurities.
High-Resolution Mass Spectrometry (HRMS): Used for identifying and confirming unknown nitrosamines, including novel NDSRIs (Nitrosamine Drug Substance-Related Impurities).
At ResolveMass Laboratories Inc., our validated nitrosamine impurity testing platform integrates all three techniques to provide orthogonal detection, confirmation, and quantification in accordance with FDA, EMA, and Health Canada guidelines.
Explore our validated nitrosamine analysis services →

While nitrate testing is distinct from nitrosamine impurity testing, it is often relevant due to the role of nitrites/nitrates in forming nitrosamines. Common methods include:
Ion Chromatography (IC): The most accurate method for quantifying nitrate (NO₃⁻) and nitrite (NO₂⁻) ions in pharmaceutical excipients and water.
UV Spectrophotometry: Applicable for nitrate detection using diazotization or Griess reagent for colorimetric analysis.
Capillary Electrophoresis: An alternative technique for nitrate/nitrite speciation, especially in complex matrices.
Monitoring nitrate content in raw materials is part of a broader nitrosamine risk assessment strategy, which complements our nitrosamine impurity testing workflows.

Products most likely to contain high concentrations of nitrosamines include:
Pharmaceuticals: Especially those containing secondary or tertiary amines in their API structure or synthesis pathway. For instance, some batches of ranitidine, sartans, and metformin have been found to contain elevated nitrosamine levels during nitrosamine impurity testing.
Cured Meats and Processed Foods: Bacon, sausages, smoked fish, and preserved meats often contain nitrosamines formed from nitrite preservatives during cooking.
Tobacco Products: Cigarettes and chewing tobacco release nitrosamines upon combustion or metabolic activation.
Rubber and Cosmetic Products: Nitrosamines may also be present in rubber nipples, condoms, or cosmetic ingredients derived from nitrosatable precursors.
At ResolveMass Laboratories Inc., our advanced nitrosamine impurity testing methods help detect trace nitrosamine levels even in complex matrices, supporting consumer safety and regulatory compliance.
Learn more about our impurity detection services →

Determination of nitrate (NO₃⁻) concentrations is crucial in both pharmaceutical and environmental analysis. Standard methods include:
Ion Chromatography (IC): The gold standard for accurate nitrate quantification in raw materials, APIs, and excipients. IC is highly selective and sensitive.
UV/Vis Spectrophotometry (Griess Method): Commonly used for nitrate and nitrite in food and water testing. Involves reduction of nitrate to nitrite followed by colorimetric detection.
Potentiometric Analysis (Ion-Selective Electrodes): Useful for rapid field testing, though it has lower specificity than chromatographic methods.
Capillary Electrophoresis (CE): Offers high resolution for nitrate/nitrite speciation, especially in multicomponent formulations.
These nitrate detection methods are often integrated into our nitrosamine risk assessment strategy, helping clients proactively prevent nitrosamine formation. This complements our confirmatory nitrosamine impurity testing protocols.
Explore our analytical solutions →

Foods that naturally contain or can form high levels of nitrosamines include:
Cured meats (e.g., bacon, hot dogs, ham): Due to nitrite preservatives that can form nitrosamines during cooking at high temperatures.
Smoked and grilled fish: Especially species like smoked salmon and mackerel.
Cheese and dairy: Some aged cheeses may accumulate trace nitrosamines during fermentation.
Beer and malt beverages: Malt drying processes may contribute to nitrosamine presence.
Pickled vegetables and fermented foods: Under certain pH and bacterial conditions.
While food exposure is monitored by regulatory agencies, nitrosamine impurity testing is more stringent in pharmaceuticals, where even trace levels can trigger recalls or reformulations.
Understand more on our nitrosamine testing protocols →

Several high-profile drug recalls have occurred due to the detection of nitrosamine impurities above acceptable intake limits. Common examples include:
Ranitidine (Zantac®) – Recalled globally after detection of NDMA under storage and heat conditions.
Sartans (e.g., Valsartan, Losartan) – Recalled due to contamination with NDMA, NDEA, and NMBA during synthesis.
Metformin ER formulations – Recalled in various regions due to NDMA detection above safe limits.
Rifampin and Rifapentine – Under surveillance due to risk of N-nitrosamine formation.
Our validated nitrosamine impurity testing services help prevent such outcomes through early detection, confirmatory analysis, and NDSRI synthesis.
Learn more about how ResolveMass supports regulatory action →

Nitrosamines are classified as probable human carcinogens, but they do not cause immediate or acute symptoms. However, long-term exposure—even at trace levels—has been linked to:
Liver damage and fibrosis
DNA alkylation and mutation
Gastrointestinal cancer risks
Tumor development in various organs (observed in animal studies)
Since symptoms develop over prolonged exposure, nitrosamine impurity testing in pharmaceuticals is critical to avoid chronic ingestion risks.
If your product is at risk, our team at ResolveMass can provide comprehensive confirmatory testing and risk assessment services to ensure regulatory compliance and patient safety.
Explore our advanced testing capabilities →

ResolveMass Laboratories Inc. provides nitrosamine impurity testing services for Canadian pharma and biotech programs, supporting both APIs and finished dosage forms. We accept Canadian shipments with controlled logistics and deliver data packages formatted for regulatory use. Our workflows cover screening, confirmatory testing, and targeted quantitation with low-level sensitivity.

You can outsource nitrosamine impurity testing to ResolveMass Laboratories Inc. for end-to-end support—from sample intake planning to validated or qualified analytical execution. We handle routine release, development batches, and investigation samples with clear chain-of-custody. Reports are issued in a regulatory-ready format aligned to common global expectations.

ResolveMass Laboratories Inc. offers nitrosamine risk assessment support tailored for IND and NDA programs, linking process knowledge, raw materials, and degradation pathways to testing plans. We provide documented rationale, risk ranking, and proposed control strategies to match submission needs. The output is designed to be directly usable within CMC documentation.

ResolveMass Laboratories Inc. specializes in NDSRI testing by combining structure-informed targeting with high-sensitivity mass spectrometry workflows. We support both predicted and confirmed NDSRIs, including method design around challenging matrices. Our deliverables include identification support, quantitation where standards exist, and defensible reporting.

ResolveMass Laboratories Inc. provides LC–MS/MS-based nitrosamine analysis for pharmaceuticals, including trace-level quantitation and confirmatory transitions. We optimize chromatographic separation to minimize interference and strengthen specificity. Results are delivered with instrument conditions, acceptance criteria, and data traceability appropriate for audits.

ResolveMass Laboratories Inc. supports confirmatory nitrosamine testing to verify screening hits or suspected risks during development and commercial supply. We use orthogonal confirmation approaches when needed and document identity and quantitation decisions clearly. This helps teams close investigations faster and move forward confidently.

ResolveMass Laboratories Inc. provides nitrosamine testing built to align with FDA and EMA expectations, including risk-based testing strategies and robust analytical documentation. We structure methods, system suitability, and reporting to withstand regulatory scrutiny. The focus is practical compliance: clear justification, defensible data, and ready-to-file outputs.

ResolveMass Laboratories Inc. offers nitrosamine testing for both APIs and drug products, covering a wide range of dosage forms and excipient matrices. We support early development through commercial lifecycle, including comparability and troubleshooting. Testing can be configured as screening, targeted panels, or compound-specific methods.

ResolveMass Laboratories Inc. offers method development for nitrosamine impurities, including separation optimization, sensitivity tuning, and matrix-mitigation strategies. We can develop fit-for-purpose methods for development needs and scale to validation-ready protocols. Documentation includes rationale, robustness checks, and practical transfer guidance.

ResolveMass Laboratories Inc. performs nitrosamine testing for generic drug manufacturers, supporting ANDA-related needs such as routine batch testing and risk-driven confirmatory studies. We emphasize reproducibility, quick scheduling, and audit-friendly reporting. Our approach helps generics teams meet expectations without slowing supply timelines.

ResolveMass Laboratories Inc. provides nitrosamine testing with low detection limits through sensitive MS-based quantitation and careful control of background contamination. We design sample prep and chromatography to push LOQs lower while maintaining specificity. Each report clearly states achieved LOQ/LLOD and the basis for calculation.

ResolveMass Laboratories Inc. supports nitrosamine testing within stability studies, including timepoint testing plans and trend-ready reporting. We can integrate nitrosamines into broader impurity/stability programs to reduce operational complexity. Data packages are structured for stability summaries and regulatory filing use.

ResolveMass Laboratories Inc. supports nitrosamine mitigation strategy development by connecting risk assessment findings to practical controls in process and formulation. We help identify likely formation routes, propose monitoring points, and recommend verification testing. The result is an actionable roadmap, not just a theoretical risk statement.

ResolveMass Laboratories Inc. is a North America-based option for outsourcing nitrosamine testing, with coordinated sample logistics across the region. We support multi-site teams with consistent methods, standardized reporting, and predictable turnaround. This makes it easier to manage programs spanning the US and Canada.

ResolveMass Laboratories Inc. handles nitrosamine root cause investigations by combining targeted analytical testing with process/material review to pinpoint credible sources. We support hypothesis testing (e.g., reagents, solvents, quench steps, packaging, water systems) using data-driven experiments. Findings are documented in a way teams can use for CAPA and regulatory responses.

ResolveMass Laboratories Inc. provides regulatory-ready nitrosamine analytical reports with clear methods, results, acceptance criteria, and traceable raw-data references. We present conclusions conservatively and document any deviations or limitations transparently. The format is designed to drop into CMC sections with minimal rework.

ResolveMass Laboratories Inc. offers multi-analyte methods that can test for multiple nitrosamines in a single run when the matrix and chemistry allow. We prioritize chromatographic resolution and ion-transition selectivity to avoid false positives. This approach reduces cost and timelines while maintaining defensibility.

ResolveMass Laboratories Inc. offers nitrosamine testing for biologics and peptides with workflows adapted for complex matrices and higher risk of interferences. We tailor sample preparation and cleanup to protect sensitivity and specificity. Deliverables include matrix-specific method notes and results suitable for development and regulatory contexts.

ResolveMass Laboratories Inc. supports nitrosamine testing during process changes by designing comparability testing plans that reflect the change scope and risk. We can test pre- and post-change batches, intermediates, and key raw materials as needed. Reporting highlights meaningful differences and supports change-control documentation.

ResolveMass Laboratories Inc. offers fast-turnaround nitrosamine testing options for urgent investigations, release needs, and time-sensitive development decisions. We prioritize rapid sample login, clear communication on feasibility, and streamlined reporting. When speed matters, we focus on delivering reliable answers without cutting corners on data integrity.