Nitrosamine Drug Substance-Related Impurities (NDSRIs)

Nitrosamine Drug Substance-Related Impurities (NDSRIs) in Pharmaceutical Formulation

The discovery of Nitrosamine Drug Substance-Related Impurities (NDSRIs) in pharmaceuticals has sent ripples through the industry, prompting stringent regulatory actions and a reevaluation of manufacturing processes. These impurities, tied directly to the chemical structure of active pharmaceutical ingredients (APIs), pose a unique challenge due to their potential carcinogenicity. Guided by frameworks like USP <1469> and the European Medicines Agency (EMA), the pharmaceutical sector is tackling NDSRIs with advanced analytics, risk assessments, and mitigation strategies. This article dives deep into the chemistry of NDSRIs, their sources, regulatory controls, and real-world examples involving drugs like valsartan, ranitidine, metformin, losartan, and nizatidine.

What Are NDSRIs?

  • NDSRIs are a subset of nitrosamine impurities formed when secondary or tertiary amine groups in an API or its intermediates react with nitrosating agents, such as nitrites, under specific conditions.
  • Unlike general nitrosamines, which may stem from contaminated solvents or reagents, NDSRIs are structurally linked to the drug substance, making them particularly challenging to control.
  • According to USP <1469>, NDSRIs are defined as nitrosamines derived from the API or its degradation products, often unique to the drug’s chemical makeup.
  • The EMA further notes that NDSRIs can form during manufacturing, storage, or even in vivo, complicating risk management.

Common NDSRIs include:

  • N-Nitrosodimethylamine (NDMA): Found in ranitidine and metformin.
  • N-Nitrosodiethylamine (NDEA): Detected in valsartan and other sartans.
  • N-Nitroso-N-methyl-4-aminobutyric acid (NMBA): Identified in losartan.
  • N-Nitrosodiisopropylamine (NDIPA): Linked to irbesartan.

Chemistry of NDSRI Formation:

NDSRIs form through a nitrosation reaction, where a nitrosating agent (e.g., nitrous acid, NO⁺) reacts with a secondary or tertiary amine in the API. The general reaction is:

R₂NH + NO⁺ → R₂N-NO + H⁺ (Secondary amine forming a stable nitrosamine)

Secondary Amines: These are highly reactive, forming stable nitrosamines like NDMA or NDEA. For example, ranitidine’s dimethylamino group can react with nitrites to form NDMA.

Tertiary Amines: These may degrade into secondary amines under certain conditions (e.g., heat, acidic pH), indirectly forming nitrosamines. For instance, an API intermediate containing triethylamine may break down to diethylamine, leading to NDEA.

Conditions favoring nitrosation include low pH (3–5), the presence of nitrites/nitrates, and mild temperatures, as noted in USP <1469>.

Why are NDSRIs a concern?

NDSRIs are classified as probable or possible human carcinogens by the International Agency for Research on Cancer (IARC), based on animal studies linking them to liver, kidney, and stomach cancers. Their concerns include:

  • Mutagenicity: NDSRIs can alkylate DNA, increasing cancer risk over prolonged exposure.
  • API-Specificity: Their formation is tied to the drug’s structure, requiring tailored mitigation strategies.
  • Low-Level Detection: Regulatory limits (e.g., 96 ng/day for NDMA) demand highly sensitive analytical methods, as outlined in USP <1469>.
  • In Vivo Formation: The EMA warns that NDSRIs may form post-administration if APIs with amine groups encounter dietary nitrites in the stomach.

Sources of NDSRIs in Pharmaceuticals

NDSRIs originate from API-related factors, as detailed in USP <1469> and EMA guidelines:

Source Description Example
API Structure APIs with secondary/tertiary amines (e.g., piperidine, morpholine) react with nitrosating agents. Ranitidine’s dimethylamino group forms NDMA.
Degradation Products API degradation during storage or processing releases amines that form NDSRIs. Nizatidine degrades to NDMA under heat.
Manufacturing Nitrite-containing reagents or contaminated intermediates introduce nitrosating agents. Valsartan synthesis using contaminated solvents.
Packaging Nitrocellulose-based coatings or rubber stoppers release nitrosating compounds. Metformin’s NDMA linked to packaging leachables.

Real-World Pharmaceutical Examples:

NDSRIs have been detected in several widely used drugs, leading to recalls and reformulations. Below are detailed case studies:

1. Valsartan (Angiotensin II Receptor Blocker)

Issue: In 2018, NDMA and NDEA were detected in valsartan batches due to a change in synthesis using a solvent (dimethylformamide, DMF) contaminated with dimethylamine.

The API’s tetrazole ring formation involved sodium nitrite, enabling nitrosation.

Impact: Global recalls were issued, affecting millions of patients. The EMA estimated NDMA levels in some batches exceeded 0.3 ppm, above the acceptable limit.

Action: Manufacturers revised synthesis processes, eliminating nitrite-containing reagents.

2. Ranitidine (H2 Receptor Antagonist)

Issue: Ranitidine’s API contains a dimethylamino group, which can self-degrade to form NDMA, especially under heat or acidic conditions. Levels as high as 0.5 ppm were detected.

Impact: The FDA and EMA recalled ranitidine in 2019–2020, citing unacceptable NDMA levels.

Action: Many manufacturers discontinued ranitidine, replacing it with alternatives like famotidine.

3. Metformin (Antidiabetic)

Issue: NDMA was found in metformin due to nitrite contamination in excipients and packaging leachables. Levels were typically low (e.g., 0.02–0.05 ppm) but concerning for chronic use.

Impact: Limited recalls occurred, with the EMA requiring enhanced testing.

Action: Manufacturers implemented stricter excipient screening and packaging controls.

4. Losartan (Angiotensin II Receptor Blocker)

Issue: NMBA was detected in losartan due to a byproduct of butyric acid derivatives in synthesis. Levels reached 0.9 ppm in some batches.

Impact: Recalls were issued in 2019, as NMBA exceeded the EMA’s interim limit of 0.96 ppm.

Action: Process optimization eliminated the problematic intermediate.

5. Nizatidine (H2 Receptor Antagonist)

Issue: Similar to ranitidine, nizatidine’s API contains a secondary amine, leading to NDMA formation during storage, particularly under high humidity.

Impact: Recalls were issued in 2020 after NDMA levels exceeded 0.3 ppm.

Action: Stability studies were enhanced, and storage conditions were tightened.

Regulatory Framework: USP <1469> and EMA Guidelines

Regulatory bodies provide robust frameworks to manage NDSRIs, ensuring patient safety.

USP <1469>: Nitrosamine Impurities

USP General Chapter <1469> outlines a comprehensive approach to nitrosamine control, including NDSRIs:

  • Risk Assessment: Evaluate APIs for amine-containing structures and nitrosation risks using tools like chemical structure analysis.
  • Analytical Methods: Use high-sensitivity techniques like LC-MS/MS (limit of detection ~0.1 ppb) or GC-MS for volatile NDSRIs.
  • Control Strategies: Implement process controls (e.g., avoiding sodium nitrite) and monitor degradation during stability testing.

EMA Guidelines

The EMA’s “Questions and Answers on Nitrosamine Impurities” mandates a three-step process:

  1. Risk Evaluation: Screen APIs for secondary/tertiary amines and assess manufacturing processes for nitrosation risks.
  2. Confirmatory Testing: Quantify NDSRIs using validated methods if risks are identified.
  3. Mitigation: Reformulate APIs, adjust synthesis conditions, or enhance supply chain controls to keep NDSRIs below AI limits.

The EMA sets AI limits, such as 96 ng/day for NDMA and 26.5 ng/day for NDEA, with case-by-case limits for novel NDSRIs.

Calculating NDSRI Limits

Calculating acceptable NDSRI levels involves the Acceptable Intake (AI) and Maximum Daily Dose (MDD), as per USP <1469> and EMA:

Acceptable Concentration (ng/g) = AI (ng/day) ÷ MDD (g/day)

Example Calculations:

  1. Valsartan (NDMA):
    • AI: 96 ng/day
    • MDD: 0.32 g/day (320 mg)
    • Limit = 96 ÷ 0.32 = 300 ng/g (0.3 ppm)
  2. Ranitidine (NDMA):
    • AI: 96 ng/day
    • MDD: 0.3 g/day (300 mg)
    • Limit = 96 ÷ 0.3 = 320 ng/g (0.32 ppm)
  3. Losartan (NMBA):
    • AI: 96 ng/day (interim)
    • MDD: 0.1 g/day (100 mg)
    • Limit = 96 ÷ 0.1 = 960 ng/g (0.96 ppm)

For multiple NDSRIs, the EMA’s “Sum of Ratios” approach (per ICH M7) ensures combined risk is acceptable:

Sum of (Ci / AIi) ≤ 1, where Ci is the concentration of NDSRI i, and AIi is its acceptable intake.

Analytical Methods for NDSRI Detection

Detecting NDSRIs requires high-sensitivity methods, as outlined in USP <1469>:

  • LC-MS/MS: Preferred for non-volatile NDSRIs (e.g., NMBA) with detection limits below 0.1 ppb.
  • GC-MS: Ideal for volatile NDSRIs like NDMA and NDEA.
  • HPLC-UV: Used for specific NDSRIs with distinct chromophores, though less sensitive.

Example Workflow:

  1. Extract NDSRIs using liquid-liquid extraction with dichloromethane.
  2. Analyze samples via LC-MS/MS, comparing peak areas to calibration standards.
  3. Calculate concentration: Concentration (ng/g) = (Peak Area ÷ Calibration Slope) × (Dilution Factor ÷ Sample Weight).

Mitigating NDSRI Risks

Both USP <1469> and EMA recommend proactive mitigation strategies:

  • Process Optimization: Use nitrite-free reagents and maintain neutral pH during synthesis (e.g., valsartan process changes).
  • Supply Chain Audits: Screen raw materials for nitrites/amines (e.g., metformin excipient controls).
  • Stability Testing: Conduct accelerated stability studies to monitor NDSRI formation (e.g., ranitidine storage adjustments).
  • Packaging Controls: Use low-risk materials to avoid leachables (e.g., metformin packaging upgrades).

Implications for Patients and Industry:

  • For patients, NDSRIs pose a low but cumulative risk, particularly for chronic medications like metformin or losartan. Recalls, such as those for ranitidine and valsartan, underscore the need for vigilance.
  • Patients should consult healthcare providers before stopping medications, as untreated conditions (e.g., hypertension) may pose greater risks.

For the industry, NDSRIs have driven significant changes:

  • Cost Increases: Enhanced testing and process optimization raise production costs.
  • Regulatory Compliance: Adherence to USP <1469> and EMA guidelines requires investment in analytics and training.
  • Innovation: Companies are developing nitrite-free synthesis routes and novel packaging solutions.

Looking Ahead:

  • NDSRIs highlight the complexity of ensuring pharmaceutical safety in a globalized supply chain.
  • Advances in analytical technology, such as ultra-high-resolution mass spectrometry, and regulatory harmonization via USP <1469> and EMA guidelines are improving NDSRI management.
  • Ongoing research into in vivo nitrosation and API stability will further refine mitigation strategies, ensuring safer drugs for patients worldwide.

References

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