Dr. BM Rao - Presentation

NITROSAMINE IMPURITIES IN API AND PHARMACEUTICAL
PRODUCTS – HOW TO DEAL?

Dr. BM Rao, Ph.D.
Head – EM QA, ASAT & Corporate Quality control

Dr. Reddy’s Laboratories, Hyderabad, India

Dr. Reddy’s Laboratories Ltd. 1

DISCLAIMER

The views expressed in this presentation are the speaker’s
personal views and do not represent the views of their current
and past employers.

Dr. Reddy’s Laboratories Ltd. 2

AGENDA

• Guidance documents
• List of Nitrosamines
• Potential sources
• Risk assessment
• Recommended methods
• Practical Analytical challenges
• Robust Approach.

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GUIDANCE DOCUMENTS FOR NITROSAMINE IMPURITIES

European Medicines Agency Information on nitrosamines for marketing authorisation
EMA/189634/2019; holders.
19 September 2019
Information to Marketing Authorisation Holders (MAHs) of
Health Canada Human Pharmaceutical Products regarding Nitrosamine
October 2, 2019
Impurities.

FDA has published testing methods Combined Direct injection N-Nitrosodimethylamine (NDMA)
and Nitrosodiethylamine (NDEA) impurity Assay by GC/MS.

European Medicines Agency Question and answers on “Information on nitrosamines for
EMA/CHMP/428592/2019 Rev.2 marketing authorisation holders”

20 December 2019

USP <381> and <1664.1> Nitrosamines in Elastomers
General Chapter Prospectus: <1469> Nitrosamine Impurities (posted on 24-Apr-2020,
Input Deadline : 22-May-2020)

APIC, 2020 Active Pharmaceutical Ingredients Committee (APIC)

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NITROSAMINES

INTRODUCTION POTENTIAL ISSUE AND IMPACT

• Nitrosamines, or more correctly N-nitrosoamines, refer • In 2018, nitrosamines were found in a number of

to any molecule containing the nitroso functional angiotensin II receptor blockers (blood pressure

group bonded to a deprotonated amine medications) known as ‘sartans’
• These molecules are of concern because of probable • Since then, nitrosamine impurity has been detected

carcinogenic nature in humans in a few batches of sartans and ranitidine
• Worldwide review of various sartans and ranitidine

• Although there are reports of its presence in some products has been initiated

foods and drinking water supplies, their presence in • This issue led various recall of several products
medicines is considered unacceptable from USA and EU markets

• Regulators have now stringent manufacturing

requirements for these medicines

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POTENTIAL SOURCE OF NITROSAMINE IMPURITIES IN API AND DP
What are the currently identified root causes for presence of nitrosamines?

Primary packing Use of Sodium Use of
materials contaminated
nitrite (NaNO2) in raw materials in
blisters in which the
manufacturing the API mfg.
“nitro cellulosic” process process (e. g:
solvents, reagents
lidding foil reacts with and catalysts)
amines in the printing
primer used to form

nitrosamines.

Degradation Nitrosamines
processes of
Use of recovered
starting materials (e. g:
materials, solvents, reagents
intermediates
and finished and catalysts)
products

Cross Use of
contaminations contaminated

due to different starting
process run on the materials and
intermediates
same line.

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LIST OF NITROSAMINES

S. no Name of the impurity Structure

1 N-nitrosodimethylamine (NDMA) Dr. Reddy’s Laboratories Ltd. 7

2 N-nitrosodiethylamine (NDEA)

3 N-nitroso-N-methylamino butyric acid (NMBA)
4 N-nitroso-di-n-butylamine (NDBA)

5 N-ethyl-N-nitroso-2-propanamine (NENPA)

6 N-nitroso-diisopropylamine (NDIPA)
7 N-isopropyl-N-ethyl nitrous amine (EIPNA)

8 N-Nitrosomorpholine (NMOR)
9 N-Nitrosopiperidine (NPIP)
10 N-Nitrosopyrrolidine (NPYR)

ORIGIN AND CHEMICAL STRUCTURE OF NITROSAMINES IN SARTANS

• To produce nitrosamine presence of
secondary amine and nitrite (under acidic
pH) required

• Risk assessment need to performed based
on the chemicals used during the
manufacturing process

• Based on risk assessment nitrosamine
impurities – selected for the analysis

Ref: Oliver et.al JPBA 172 (2019) 278-284

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EDQM GUIDANCE TO AVOID NITROSAMINES IN HUMAN MEDICINES

• Conduct a risk evaluation to identify products at risk of N-nitrosamine
formation or (cross) contamination and report the out come by 26March

STEP-1 2020 *

• Perform further confirmatory testing on the products identified to be at

Step-2 risk of N-nitrosamine formation or (cross-) contamination and report

confirmed presence of nitrosamines as soon as possible.

• Apply for any necessary changes to the manufacturing process

Step-3 resulting from this review using the established regulatory procedures.

* The European medicine regulatory network has agreed to extent the deadline to
complete step 1 risk evaluation up to 1 October 2020.

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RISK ASSESSMENT & CONTROL

Risk identification Risk reduction /
Establishing control
Step-1 Risk Analysis Step-2
strategy
Risk Evaluation Risk Acceptance
RISK IDENTIFICATION
• Assessment of various input materials for the possible formation of nitrosamines Dr. Reddy’s Laboratories Ltd. 10
• Process assessment

RISK ANALYSIS
Nitrosamines may generate from the following materials in drug product

• Active Pharmaceutical Ingredient (Drug Substance)
• Excipients used in the manufacturing of drug product
• Packaging components used in the manufacturing of drug product
• Water used in Drug product manufacturing
• Drug product manufacturing process
• Cleaning agents used in drug product manufacturing site
• Drug product manufacturing facility (shared equipment)

RISK ASSESSMENT & CONTROL

RISK EVALUATION
• Through a FMEA – type tool with different scores assigned to various risk levels
• Through a Yes / No Questionnaire which has the benefit of better orienting the result
RISK CONTROL
Based on the outcome of risk evaluation, an appropriate control strategy shall be established in the drug product.

• Arriving at the control limits and revision of drug product specification to include a limit
for the nitrosamine impurity.

• Confirmatory testing of the drug product batches for nitrosamine impurities.
• Testing of each batch of DP as part of release testing.
• Adopting a reduced testing program for drug product batches.
• Monitoring of stability batches for nitrosamine impurities.

• At least 3 consecutive commercial batches of API should be tested to confirm the level of nitrosamines in the API.
• Whenever values above 30% of the interim limit are confirmed, Marketing Authorisation Holder should be informed.

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ANALYTICAL CONTROL CAN BE A CHALLENGE

• To Develop rugged and robust method
for quantification at low levels

• The limits for these impurities were too
low. Hence high sensitive methods
required

• Analyte should be stable in the solution
• Should be easy to adopt in Quality

Control

As testing methods have become more sophisticated and sensitive, the FDA and industry
can identify and mitigate risks to patients.

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METHODS

FDA published methods

• Combined headspace method: a GC/MS method that allows determination of both N-
Nitrosodimethylamine (NDMA) and N-Nitrosodiethylamine (NDEA) simultaneously

• Combined direct injection method: a GC-MS/MS method that allows for determination of both NDMA and
NDEA simultaneously

• Direct injection GC-MS method: a method that can detect NDMA, NDEA, N-Nitrosodiisopropylamine
(NDIPA), N-Nitrosoethylisopropylamine (NEIPA), and N-nitrosodibutylamine (NDBA)

• Headspace GC-MS method: a method that can detect NDMA, NDEA, NDIPA, and NEIPA
• LC-HRMS method: a method that can detect NDMA, NDEA, NEIPA, NDIPA, NDBA, and N-Nitroso-N-

methyl-4-aminobutyric acid (NMBA)
• Rapid Fire-MS/MS method: a method that can detect NEIPA, NDIPA, NDBA, and NMBA. We do not

recommend using this method to detect NDMA or NDEA because it is less sensitive to those impurities.

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CHALLENGES IN DEVELOPMENT OF METHODS

Efforts to identify the suitable method for the determination of Nitrosamines

• Development of sensitive methods
• Ensure the suitability of the method for target determination
• Solubility variation between analyte and impurities
• Separation and interference of other impurities
• Placebo interferences
• Saturation of detector
• Reproducibility of detection and quantitation limits

Practical analytical challenges for determination of Nitrosamines

Method development:

• Peak shape of the Nitrosamine varies with respect to the organic / aqueous ratio.
• Achieving the sensitivity of the method with respect to current limits in the API or drug products.
• However companies to make the necessary changes to their manufacturing processes and to put in place testing

regimes able to detect the smallest amount of these impurities(<0.03ppm) (Refer EMA/248364/2019 Rev 1)
• LC-MS source contamination due to usage of higher analyte (API or drug product) concentration.
• HPLC or UPLC selection as front end for the peak profile and consistency at low level. Interpretation and reporting

of results by linearity(broad range) method is a challenge.
• Handling of insoluble API’s to achieve the lower LOQ values and recovery factor of nitrosoamines is challenge in

development of method.

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CHALLENGES IN DEVELOPMENT OF METHODS

Practical analytical challenges for determination of Nitrosamines

Drug product analysis:

• Drug product sample needs to be filtered with syringe filters and achieving recovery
• Analyzing different label claim drug products by using single method.
• LOQ recovery of Nitrosoamine impurity in drug product is a challenge
• Separation of nitrosoamine impurity peak with different excipient interference

Suggested robust approach:

• Comprehensive assessment of synthetic routes and toxicological attributes
• Risk assessment includes all steps, reagents, manufacturing approaches etc.,
• Development of robust advance analytical techniques to detect potential impurities complemented and

verified orthogonal analytical techniques
• Development of sensitive methods
• Use of hyphenated techniques for quantification
• Extended linearity range
• Conduct Precision and Recovery experiments at LOQ level
• Screening various column chemistries for improved separation between analyte and product peak.
• Ensuring validation and transfer of analytical method as per the guidelines and SOP’s

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