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Updated Information | Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs)

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Recommended Acceptable Intake (AI) Limits and Testing Methods

On 8/4/2023, FDA issued a final guidance on Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs) (August 2023) (NDSRI Guidance).  To reflect the evolving and highly technical nature of the relevant information, FDA is providing certain updated NDSRI-specific information on this website in connection with the guidance.  

Specifically, FDA intends to include on this website updated information on: (1) recommended AI limits for certain NDSRIs based on their predicted carcinogenic potency categorization listed by APIs that hypothetically could be at risk of forming such NDSRIs; (2) recommended AI limits for certain NDSRIs based on compound-specific data or read-across analysis from a surrogate; (3) recommended interim AI limits for certain NDSRIs; (4) recommended testing methods for confirmatory testing of certain NDSRIs; and (5) recommended safety testing methods for NDSRIs.

This guidance (including the information on this associated website) represents the current thinking of the Food and Drug Administration (FDA or Agency) on this topic.  It does not establish any rights for any person and is not binding on FDA or the public.  You can use an alternative approach if it satisfies the requirements of the applicable statutes and regulations. 

The public may comment on the information below by submitting a comment to the public docket established for the final guidance on Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs) (Docket No. FDA-2020-D-1530).  Comments may be submitted at any time for Agency consideration.  Submit written comments to the Dockets Management Staff (HFA-305), Food and Drug Administration, 5630 Fishers Lane, Rm. 1061, Rockville, MD 20852.  Submit electronic comments to www.regulations.gov.  All comments should be identified with the docket number FDA-2020-D-1530 and complete title of the guidance.  For additional information, please refer to the Federal Register Notice of Availability for the final NDSRI Guidance, available at www.regulations.gov.


Recommended AI Limits for Certain NDSRIs

Based on Predicted Carcinogenic Potency Categorization for APIs at Hypothetical Risk of Forming Such NDSRIs

APIs that contain secondary amine or dimethyl tertiary amine centers are at risk of forming NDSRIs by nitrosation of the amine center.  This can occur under conditions related to the formulation and manufacturing process for the drug product, such as by reaction with residual nitrites in excipients used to formulate the drug product. Table 1, below, presents NDSRIs that can hypothetically form by this process from a number of amine-containing APIs.  Recommended AI limits for these hypothetical NDSRIs based on the predicted carcinogenic potency categorization approach are also presented.  If an NDSRI currently listed in Table 1 is formed, the corresponding recommended AI limit in Table 1 based on the predicted carcinogenic potency categorization approach applies.  FDA may move an NDSRI to Table 2 if compound-specific data or read-across analysis from a surrogate becomes available and an updated recommended AI limit applies.  The recommended AI limits based on the predicted carcinogenic potency characterization approach should not be applied to NDSRIs in circumstances in which FDA otherwise recommends an AI limit (e.g., based on compound-specific assessments or read-across analysis from a surrogate) in Table 2.

For APIs with more than one nitrosatable amine center, the predicted carcinogenic potency category for each unique NDSRI formed by mono-nitrosation of the API is presented in Table 1, where each NDSRI from the same API is differentiated by the suffix “-1,” “-2,” etc. in the NDSRI Name column.  Chemical structure images* for NDSRIs in Table 1 can be accessed by clicking on the hyperlinked NDSRI Name.  This table will be updated periodically based on new information.

This is not an all-inclusive list of APIs that have the potential to form NDSRIs; there are other possible routes of NDSRI formation.  In addition, the recommended AI limits for the NDSRIs on this website do not necessarily correspond to the observed level of any NDSRI(s) found in any particular drug product.  Furthermore, inclusion of an API on the list is not confirmation that an NDSRI is present in a drug product containing that drug substance.  A recommended AI limit is based on a safety assessment that includes evaluation of the mutagenic and carcinogenic potential of the impurity and represents the level at or below which FDA has determined that the impurity or impurities would not pose a safety concern for patients taking the drug product.

* Chemical structure images provided by the National Institutes of Health/National Center for Advancing Translational Sciences.

Table 1: FDA Recommended AI Limits for Certain Hypothetical NDSRIs 

Based on Predicted Carcinogenic Potency Categorization

(last updated 12/1/2023)

*Source is the form of the drug substance in the free base or free acid form.
NDSRI Name Source* Potency Category Recommended AI Limit
N-nitroso-abacavir Abacavir 5 1500 ng/day
N-nitroso-acarbose Acarbose 5 1500 ng/day
N-nitroso-acebutolol Acebutolol 4 1500 ng/day
N-nitroso-albuterol Albuterol 5 1500 ng/day
N-nitroso-desmethyl-almotriptan Almotriptan 1 26.5 ng/day
N-nitroso-desmethyl-amitriptyline Amitriptyline 1 26.5 ng/day
N-nitroso-amlodipine Amlodipine 5 1500 ng/day
N-nitroso-amoxapine Amoxapine 3 400 ng/day
N-nitroso-arformoterol Arformoterol 4 1500 ng/day
N-nitroso-argatroban Argatroban 4 1500 ng/day
N-nitroso-articaine Articaine 4 1500 ng/day
N-nitroso-atenolol Atenolol 4 1500 ng/day
N-nitroso-avacopan Avacopan 5 1500 ng/day
N-nitroso-avanafil Avanafil 3 400 ng/day
N-nitroso-desmethyl-azithromycin Azithromycin 4 1500 ng/day
N-nitroso-desmethyl-bedaquiline Bedaquiline 1 26.5 ng/day
N-nitroso-belumosudil Belumosudil 5 1500 ng/day
N-nitroso-benazepril Benazepril 5 1500 ng/day
N-nitroso-bendroflumethiazide Bendroflumethiazide 5 1500 ng/day
N-nitroso-benzonatate Benzonatate 3 400 ng/day
N-nitroso-berotralstat Berotralstat 2 100 ng/day
N-nitroso-betaxolol Betaxolol 4 1500 ng/day
N-nitroso-bicisate Bicisate 4 1500 ng/day
N-nitroso-bisoprolol Bisoprolol 4 1500 ng/day
N-nitroso-brilliant blue g Brilliant Blue G 5 1500 ng/day
N-nitroso-brinzolamide Brinzolamide 2 100 ng/day
N-nitroso-desmethyl-brompheniramine Brompheniramine 1 26.5 ng/day
N-nitroso-bumetanide Bumetanide 4 1500 ng/day
N-nitroso-bupropion Bupropion 5 1500 ng/day
N-nitroso-desmethyl-cabergoline Cabergoline 1 26.5 ng/day
N-nitroso-cangrelor Cangrelor 3 400 ng/day
N-nitroso-desmethyl-carbinoxamine Carbinoxamine 1 26.5 ng/day
N-nitroso-carteolol Carteolol 5 1500 ng/day
N-nitroso-carvedilol Carvedilol 3 400 ng/day
N-nitroso-caspofungin Caspofungin 4 1500 ng/day
N-nitroso-desmethyl-chlophedianol Chlophedianol 1 26.5 ng/day
N-nitroso-chloroquine Chloroquine 5 1500 ng/day
N-nitroso-desmethyl-chlorpheniramine Chlorpheniramine 1 26.5 ng/day
N-nitroso-desmethyl-chlorpromazine Chlorpromazine 1 26.5 ng/day
N-nitroso-cinacalcet Cinacalcet 3 400 ng/day
N-nitroso-ciprofloxacin Ciprofloxacin 4 1500 ng/day
N-nitroso-desmethyl-citalopram Citalopram 1 26.5 ng/day
N-nitroso-desmethyl-clarithromycin Clarithromycin 4 1500 ng/day
N-nitroso-clevidipine Clevidipine 5 1500 ng/day
N-nitroso-desmethyl-clomipramine Clomipramine 1 26.5 ng/day
N-nitroso-clozapine Clozapine 5 1500 ng/day
N-nitroso-colistin a hydrogen methanesulfonate-1 Colistin 2 100 ng/day
N-nitroso-colistin a hydrogen methanesulfonate-2 Colistin 2 100 ng/day
N-nitroso-colistin a hydrogen methanesulfonate-3 Colistin 2 100 ng/day
N-nitroso-colistin a hydrogen methanesulfonate-4 Colistin 2 100 ng/day
N-nitroso-colistin a hydrogen methanesulfonate-5 Colistin 2 100 ng/day
N-nitroso-colistin b hydrogen methanesulfonate-1 Colistin 2 100 ng/day
N-nitroso-colistin b hydrogen methanesulfonate-2 Colistin 2 100 ng/day
N-nitroso-colistin b hydrogen methanesulfonate-3 Colistin 2 100 ng/day
N-nitroso-colistin b hydrogen methanesulfonate-4 Colistin 2 100 ng/day
N-nitroso-colistin b hydrogen methanesulfonate-5 Colistin 2 100 ng/day
N-nitroso-desmethyl-cyclobenzaprine Cyclobenzaprine 1 26.5 ng/day
N-nitroso-dabigatran etexilate Dabigatran Etexilate 3 400 ng/day
N-nitroso-degarelix Degarelix 3 400 ng/day
N-nitroso-desmethyl-demeclocycline Demeclocycline 3 400 ng/day
N-nitroso-desipramine Desipramine 1 26.5 ng/day
N-nitroso-desloratadine Desloratadine 3 400 ng/day
N-nitroso-desmethyl-desvenlafaxine Desvenlafaxine 1 26.5 ng/day
N-nitroso-deucravacitinib Deucravacitinib 5 1500 ng/day
N-nitroso-desmethyl-dexbrompheniramine Dexbrompheniramine 1 26.5 ng/day
N-nitroso-desmethyl-dexchlorpheniramine Dexchlorpheniramine 1 26.5 ng/day
N-nitroso-diclofenac Diclofenac 5 1500 ng/day
N-nitroso-desmethyl-diltiazem Diltiazem 1 26.5 ng/day
N-nitroso-desmethyl-diphenhydramine Diphenhydramine 1 26.5 ng/day
N-nitroso-dipivefrin Dipivefrin 2 100 ng/day
N-nitroso-dobutamine Dobutamine 4 1500 ng/day
N-nitroso-dorzolamide Dorzolamide 2 100 ng/day
N-nitroso-desmethyl-cidoxepin Doxepin 1 26.5 ng/day
N-nitroso-desmethyl-doxepin, (e)- Doxepin 1 26.5 ng/day
N-nitroso-desmethyl-doxycycline Doxycycline 3 400 ng/day
N-nitroso-desmethyl-doxylamine Doxylamine 1 26.5 ng/day
N-nitroso-duvelisib Duvelisib 5 1500 ng/day
N-nitroso-elagolix Elagolix 4 1500 ng/day
N-nitroso-enalapril Enalapril 5 1500 ng/day
N-nitroso-enalaprilat Enalaprilat 5 1500 ng/day
N-nitroso-ephedrine Ephedrine 4 1500 ng/day
N-nitroso-epinephrine Epinephrine 2 100 ng/day
N-nitroso-desmethyl-eravacycline Eravacycline 3 400 ng/day
N-nitroso-ertapenem Ertapenem 4 1500 ng/day
N-nitroso-desmethyl-erythromycin Erythromycin 4 1500 ng/day
N-nitroso-desmethyl-erythromycin ethylsuccinate Erythromycin Ethylsuccinate 3 400 ng/day
N-nitroso-desmethyl-escitalopram Escitalopram 1 26.5 ng/day
N-nitroso-esketamine Esketamine 5 1500 ng/day
N-nitroso-esmolol Esmolol 4 1500 ng/day
N-nitroso-ethambutol Ethambutol 4 1500 ng/day
N-nitroso-etravirine Etravirine 5 1500 ng/day
N-nitroso-exametazime Exametazime 4 1500 ng/day
N-nitroso-felodipine Felodipine 5 1500 ng/day
N-nitroso-fenfluramine Fenfluramine 3 400 ng/day
N-nitroso-fenoldopam Fenoldopam 2 100 ng/day
N-nitroso-finerenone Finerenone 5 1500 ng/day
N-nitroso-flecainide Flecainide 4 1500 ng/day
N-nitroso-florbetaben f-18 Florbetaben F-18 2 100 ng/day
N-nitroso-florbetapir f-18 Florbetapir F-18 2 100 ng/day
N-nitroso-flutemetamol f-18 Flutemetamol F-18 2 100 ng/day
N-nitroso-folic acid Folic Acid 4 1500 ng/day
N-nitroso-formoterol Formoterol 4 1500 ng/day
N-nitroso-fosdenopterin-1 Fosdenopterin 5 1500 ng/day
N-nitroso-fosdenopterin-2 Fosdenopterin 5 1500 ng/day
N-nitroso-fostamatinib-1 Fostamatinib 5 1500 ng/day
N-nitroso-fostamatinib-2 Fostamatinib 5 1500 ng/day
N-nitroso-frovatriptan Frovatriptan 3 400 ng/day
N-nitroso-furosemide Furosemide 4 1500 ng/day
N-nitroso-gatifloxacin Gatifloxacin 4 1500 ng/day
N-nitroso-hydrochlorothiazide Hydrochlorothiazide 4 1500 ng/day
N-nitroso-hydroxychloroquine Hydroxychloroquine 5 1500 ng/day
N-nitroso-imatinib Imatinib 5 1500 ng/day
N-nitroso-desmethyl-imipramine Imipramine 1 26.5 ng/day
N-nitroso-isoproterenol Isoproterenol 4 1500 ng/day
N-nitroso-isradipine Isradipine 5 1500 ng/day
N-nitroso-ivacaftor Ivacaftor 5 1500 ng/day
N-nitroso-ketamine Ketamine 5 1500 ng/day
N-nitroso-labetalol Labetalol 4 1500 ng/day
N-nitroso-leniolisib Leniolisib 5 1500 ng/day
N-nitroso-leucovorin-1 Leucovorin 4 1500 ng/day
N-nitroso-leucovorin-2 Leucovorin 4 1500 ng/day
N-nitroso-levalbuterol Levalbuterol 5 1500 ng/day
N-nitroso-levamlodipine Levamlodipine 5 1500 ng/day
N-nitroso-levmetamfetamine Levmetamfetamine 3 400 ng/day
N-nitroso-levobunolol Levobunolol 5 1500 ng/day
N-nitroso-levoleucovorin-1 Levoleucovorin 4 1500 ng/day
N-nitroso-levoleucovorin-2 Levoleucovorin 4 1500 ng/day
N-nitroso-levomefolic acid-1 Levomefolic Acid 4 1500 ng/day
N-nitroso-levomefolic acid-2 Levomefolic Acid 4 1500 ng/day
N-nitroso-lisinopril Lisinopril 5 1500 ng/day
N-nitroso-desmethyl-maralixibat Maralixibat 2 100 ng/day
N-nitroso-maribavir Maribavir 5 1500 ng/day
N-nitroso-mecamylamine Mecamylamine 5 1500 ng/day
N-nitroso-meclofenamic acid Meclofenamic Acid 5 1500 ng/day
N-nitroso-mefloquine Mefloquine 4 1500 ng/day
N-nitroso-meropenem Meropenem 4 1500 ng/day
N-nitroso-desmethyl-methadone Methadone 3 400 ng/day
N-nitroso-methamphetamine Methamphetamine 3 400 ng/day
N-nitroso-desmethyl-methylene blue Methylene Blue 2 100 ng/day
N-nitroso-metolazone Metolazone 5 1500 ng/day
N-nitroso-metoprolol Metoprolol 4 1500 ng/day
N-nitroso-desmethyl-mifepristone Mifepristone 2 100 ng/day
N-nitroso-migalastat Migalastat 4 1500 ng/day
N-nitroso-desmethyl-minocycline-1 Minocycline 2 100 ng/day
N-nitroso-desmethyl-minocycline-2 Minocycline 3 400 ng/day
N-nitroso-mirabegron Mirabegron 3 400 ng/day
N-nitroso-mitoxantrone-1 Mitoxantrone 4 1500 ng/day
N-nitroso-mitoxantrone-2 Mitoxantrone 2 100 ng/day
N-nitroso-moexipril Moexipril 5 1500 ng/day
N-nitroso-moxifloxacin Moxifloxacin 4 1500 ng/day
N-nitroso-nadolol Nadolol 5 1500 ng/day
N-nitroso-nebivolol Nebivolol 4 1500 ng/day
N-nitroso-desmethyl-neratinib Neratinib 2 100 ng/day
N-nitroso-neratinib Neratinib 5 1500 ng/day
N-nitroso-nicardipine Nicardipine 5 1500 ng/day
N-nitroso-nifedipine Nifedipine 5 1500 ng/day
N-nitroso-nimodipine Nimodipine 5 1500 ng/day
N-nitroso-nintedanib Nintedanib 5 1500 ng/day
N-nitroso-nisoldipine Nisoldipine 5 1500 ng/day
N-nitroso-desmethyl-nizatidine Nizatidine 1 26.5 ng/day
N-nitroso-nizatidine-1 Nizatidine 2 100 ng/day
N-nitroso-nizatidine-2 Nizatidine 3 400 ng/day
N-nitroso-nortriptyline Nortriptyline 1 26.5 ng/day
N-nitroso-olanzapine Olanzapine 5 1500 ng/day
N-nitroso-oliceridine Oliceridine 1 26.5 ng/day
N-nitroso-olodaterol Olodaterol 5 1500 ng/day
N-nitroso-desmethyl-olopatadine Olopatadine 4 1500 ng/day
N-nitroso-desmethyl-omadacycline-1 Omadacycline 2 100 ng/day
N-nitroso-desmethyl-omadacycline-2 Omadacycline 3 400 ng/day
N-nitroso-omadacycline Omadacycline 1 26.5 ng/day
N-nitroso-omidenepag isopropyl Omidenepag Isopropyl 4 1500 ng/day
N-nitroso-oritavancin-1 Oritavancin 4 1500 ng/day
N-nitroso-oritavancin-2 Oritavancin 5 1500 ng/day
N-nitroso-desmethyl-orphenadrine Orphenadrine 1 26.5 ng/day
N-nitroso-ozanimod Ozanimod 3 400 ng/day
N-nitroso-ozenoxacin Ozenoxacin 4 1500 ng/day
N-nitroso-desmethyl-padimate o Padimate O 2 100 ng/day
N-nitroso-pafolacianine Pafolacianine 4 1500 ng/day
N-nitroso-perindopril Perindopril 5 1500 ng/day
N-nitroso-desmethyl-pheniramine Pheniramine 1 26.5 ng/day
N-nitroso-phenylephrine Phenylephrine 2 100 ng/day
N-nitroso-desmethyl-phenyltoloxamine Phenyltoloxamine 1 26.5 ng/day
N-nitroso-pindolol Pindolol 4 1500 ng/day
N-nitroso-plazomicin-1 Plazomicin 4 1500 ng/day
N-nitroso-plazomicin-2 Plazomicin 2 100 ng/day
N-nitroso-plerixafor-1 Plerixafor 2 100 ng/day
N-nitroso-plerixafor-2 Plerixafor 2 100 ng/day
N-nitroso-plerixafor-3 Plerixafor 2 100 ng/day
N-nitroso-polythiazide Polythiazide 5 1500 ng/day
N-nitroso-pramipexole Pramipexole 3 400 ng/day
N-nitroso-prilocaine Prilocaine 4 1500 ng/day
N-nitroso-primaquine Primaquine 5 1500 ng/day
N-nitroso-proline Proline 4 1500 ng/day
N-nitroso-desmethyl-promethazine Promethazine 3 400 ng/day
N-nitroso-propafenone Propafenone 2 100 ng/day
N-nitroso-desmethyl-propoxyphene Propoxyphene 1 26.5 ng/day
N-nitroso-propranolol Propranolol 4 1500 ng/day
N-nitroso-propylhexedrine Propylhexedrine 3 400 ng/day
N-nitroso-protriptyline Protriptyline 1 26.5 ng/day
N-nitroso-pseudoephedrine Pseudoephedrine 4 1500 ng/day
N-nitroso-desmethyl-pyrilamine Pyrilamine 1 26.5 ng/day
N-nitroso-quinapril Quinapril 5 1500 ng/day
N-nitroso-desmethyl-quinupristin Quinupristin 2 100 ng/day
N-nitroso-racepinephrine Racepinephrine 2 100 ng/day
N-nitroso-ramipril Ramipril 5 1500 ng/day
N-nitroso-desmethyl-ranitidine Ranitidine 1 26.5 ng/day
N-nitroso-ranitidine-1 Ranitidine 3 400 ng/day
N-nitroso-ranitidine-2 Ranitidine 2 100 ng/day
N-nitroso-rasagiline Rasagiline 2 100 ng/day
N-nitroso-relebactam Relebactam 3 400 ng/day
N-nitroso-rifabutin Rifabutin 5 1500 ng/day
N-nitroso-rilpivirine-1 Rilpivirine 5 1500 ng/day
N-nitroso-rilpivirine-2 Rilpivirine 5 1500 ng/day
N-nitroso-risdiplam Risdiplam 5 1500 ng/day
N-nitroso-desmethyl-rivastigmine Rivastigmine 2 100 ng/day
N-nitroso-desmethyl-rizatriptan Rizatriptan 1 26.5 ng/day
N-nitroso-rolapitant Rolapitant 5 1500 ng/day
N-nitroso-safinamide Safinamide 3 400 ng/day
N-nitroso-salmeterol Salmeterol 3 400 ng/day
N-nitroso-sapropterin-1 Sapropterin 4 1500 ng/day
N-nitroso-sapropterin-2 Sapropterin 5 1500 ng/day
N-nitroso-desmethyl-sarecycline Sarecycline 3 400 ng/day
N-nitroso-sertraline Sertraline 2 100 ng/day
N-nitroso-silodosin Silodosin 4 1500 ng/day
N-nitroso-sotalol Sotalol 4 1500 ng/day
N-nitroso-desmethyl-spinosad factor a Spinosad 2 100 ng/day
N-nitroso-desmethyl-spinosad factor d Spinosad 2 100 ng/day
N-nitroso-streptomycin Streptomycin 4 1500 ng/day
N-nitroso-desmethyl-sumatriptan Sumatriptan 1 26.5 ng/day
N-nitroso-tafenoquine Tafenoquine 5 1500 ng/day
N-nitroso-desmethyl-tamoxifen Tamoxifen 1 26.5 ng/day
N-nitroso-tamsulosin Tamsulosin 4 1500 ng/day
N-nitroso-desmethyl-tapentadol Tapentadol 1 26.5 ng/day
N-nitroso-telavancin-1 Telavancin 3 400 ng/day
N-nitroso-telavancin-2 Telavancin 4 1500 ng/day
N-nitroso-telavancin-3 Telavancin 4 1500 ng/day
N-nitroso-telavancin-4 Telavancin 5 1500 ng/day
N-nitroso-desmethyl-telithromycin Telithromycin 4 1500 ng/day
N-nitroso-terbutaline Terbutaline 5 1500 ng/day
N-nitroso-desmethyl-tetracaine Tetracaine 1 26.5 ng/day
N-nitroso-tetracaine Tetracaine 3 400 ng/day
N-nitroso-desmethyl-tetracycline Tetracycline 3 400 ng/day
N-nitroso-desmethyl-thonzylamine Thonzylamine 1 26.5 ng/day
N-nitroso-ticagrelor Ticagrelor 5 1500 ng/day
N-nitroso-desmethyl-tigecycline-1 Tigecycline 3 400 ng/day
N-nitroso-desmethyl-tigecycline-2 Tigecycline 2 100 ng/day
N-nitroso-tigecycline Tigecycline 5 1500 ng/day
N-nitroso-timolol Timolol 5 1500 ng/day
N-nitroso-tirofiban Tirofiban 4 1500 ng/day
N-nitroso-torsemide Torsemide 5 1500 ng/day
N-nitroso-desmethyl-tramadol Tramadol 1 26.5 ng/day
N-nitroso-trandolapril Trandolapril 5 1500 ng/day
N-nitroso-trientine Trientine 1 26.5 ng/day
N-nitroso-desmethyl-trimethobenzamide Trimethobenzamide 1 26.5 ng/day
N-nitroso-desmethyl-trimipramine Trimipramine 1 26.5 ng/day
N-nitroso-desmethyl-ulipristal acetate Ulipristal Acetate 2 100 ng/day
N-nitroso-vancomycin Vancomycin 4 1500 ng/day
N-nitroso-desmethyl-venlafaxine Venlafaxine 1 26.5 ng/day
N-nitroso-vibegron Vibegron 5 1500 ng/day
N-nitroso-vilanterol Vilanterol 3 400 ng/day
N-nitroso-viloxazine Viloxazine 2 100 ng/day
N-nitroso-vortioxetine Vortioxetine 3 400 ng/day
N-nitroso-desmethyl-zolmitriptan Zolmitriptan 1 26.5 ng/day

 


Recommended AI Limits for Certain NDSRIs 

Based on Compound-Specific Data or Read-Across Analysis from a Surrogate

As indicated in the NDSRI Guidance, a recommended AI limit is based on a safety assessment that includes evaluation of the mutagenic and carcinogenic potential of the impurity and represents the level at or below which FDA has determined that the impurity or impurities would not pose a safety concern for patients.  A recommended compound-specific AI limit can be calculated based on rodent carcinogenic potency data such as TD50 values identified in the published scientific literature.  When the mutagenic potential of an NDSRI is not adequately characterized, FDA has used structure activity relationships to support the identification of a robustly-tested surrogate that is similar in structure and reactivity to the NDSRI to generate an estimate of carcinogenic potency from which an AI limit can be scientifically determined.  The rationale for the choice of surrogate (similar in structure and reactivity) is significant because test data from the identified surrogate are then used to generate an estimate, either quantitatively or qualitatively, for a compound that lacks robust mutagenicity and carcinogenicity data (commonly referred to as a read-across analysis.) 

Table 2 presents FDA recommended AI Limits for certain NDSRIs based on compound-specific data or read-across analysis from a surrogate. If an NDSRI appears in Table 2, the recommended AI limit in Table 2 applies, rather than a limit based on the predicted carcinogenic potency categorization approach.

Table 2: FDA Recommended AI Limits for Certain NDSRIs 

Based on Compound-Specific Data or Read-Across Analysis from a Surrogate

(last updated 12/1/2023)

NDSRI Name Source Recommended AI Limit (ng/day) Surrogate Date Added to Table
7-Nitroso-3-(trifluoromethyl)-5,6,7,8-tetrahydro[1,2,4]triazolo-[4,3-a]pyrazine (NTTP) Sitagliptin 37 N-nitroso-1,2,3,6-tetrahydropyridine (NTHP) 8/4/2023*
N-nitroso-varenicline Varenicline 37 NTHP 8/4/2023**
N-nitroso-duloxetine Duloxetine 100 4-(methylnitrosoamino)-1-(3-pyridinyl)-1-butanone (NNK) 8/4/2023***
N-nitroso-fluoxetine Fluoxetine 100 4-(methylnitrosoamino)-1-(3-pyridinyl)-1-butanone (NNK) 10/11/2023
N-nitroso-atomoxetine Atomoxetine 100 4-(methylnitrosoamino)-1-(3-pyridinyl)-1-butanone (NNK) 12/1/2023

*This limit was previously communicated on August 9, 2022.
**This limit was previously communicated on July 16, 2021.
***See Table 3.


Recommended Interim AI Limits for Certain NDSRIs

As indicated in the NDSRI Guidance, if drug product batches already in distribution contain NDSRIs at levels above the FDA recommended AI limit, and manufacturing changes or recalls are likely to lead to a disruption in the drug supply, then manufacturers and applicants should immediately contact CDER’s Drug Shortage Staff at drugshortages@fda.hhs.gov. When contacted about a potential disruption in the drug supply, FDA intends to evaluate each circumstance on a case-by-case basis. FDA may work directly with a specific manufacturer or applicant of the marketed drug and intends to consider whether it is appropriate or not appropriate to recommend an interim AI limit for a temporary period. If FDA recommends an interim AI limit, it generally does not intend to object, for example based on applicable underlying CGMP violations, to distribution of such drug product batches that contain NDSRI levels at or below the recommended interim AI limit during the specified period under certain circumstances on a case-by-case basis. In certain cases where FDA does not intend to object to the distribution of drug products from multiple drug manufacturers that contain NDSRI levels at or below the recommended interim AI limit, FDA intends to post such recommended interim AI limits on this website.

Table 3: Recommended Interim AI Limits* for Certain NDSRIs** for Approved or Currently Marketed Products

(last updated: 2/23/2024)

NDSRI Name

Source

Recommended Interim AI Limit (ng/day)

Recommended Interim Control Limit (ppm)

Estimated Duration***

N-nitroso-duloxetine

Duloxetine

600 ng/day

5 ppm

10/1/2024

* Note that an AI limit can be converted into a parts per million (ppm) control limit. The conversion varies by product and is calculated based on a drug’s maximum daily dose (MDD) as reflected in the drug labeling (ppm = AI (nanograms (ng))/MDD (milligrams)).
**A decision to release lots remains solely the firm’s responsibility.  Firms are responsible for ensuring that their drugs are manufactured in compliance with all applicable requirements, including CGMP, and FDA expects them to vigilantly monitor and promptly report to FDA any adverse drug experiences or other findings that may affect product quality or safety.
***“Estimated Duration” is the date by which FDA intends to reassess the recommended interim control limit.  As indicated in the NDSRI guidance, while FDA recommends conclusion of NDSRI confirmatory testing of drug products and submission of required changes in drug applications by August 1, 2025 for approved or currently marketed products, FDA may recommend that firms complete an expedited risk assessment, confirmatory testing, or other regulatory action based on information available to the Agency.  With respect to FDA-recommended interim control limits, FDA generally does not intend to object to the distribution by manufacturers and applicants of drug products that contain NDSRI levels at or below the recommended interim control limit, until the date identified in the “Estimated Duration” column. However, if FDA becomes aware that the shortage risk has been alleviated, FDA may provide additional notification or may reconsider its recommended interim control limit.

Recommended Testing Methods for Confirmatory Testing of Certain NDSRIs

As indicated in the NDSRI Guidance, if manufacturers and applicants identify a risk of NDSRI formation in a drug product, then confirmatory testing of batches should be conducted using sensitive and appropriately validated methods. 

FDA intends to provide information on FDA-generated testing methods to provide options for regulators and industry to detect NDSRI impurities in specific drug substances and drug products.  The methods should be validated by the user if the resulting data are used to support a required quality assessment of the drug substance or drug product, or if the results are used in a regulatory submission.

LC-ESI-HRMS Method for the Determination of N-Nitroso-Bumetanide

LC-ESI-HRMS Method for the Determination of N-Nitroso-Propranolol

LC-ESI-HRMS Method for the Determination of N-Nitroso-Varenicline

Recommended Safety Testing Methods for NDSRIs

As indicated in the NDSRI Guidance, a manufacturer or applicant may submit a scientifically justified rationale to pursue an AI limit higher than the FDA recommended limit associated with the predicted carcinogenic potency category for that NDSRI.  Alternative approaches using safety data, such as obtaining compound-specific data or using read-across assessment to a suitable surrogate, could be used to support a higher limit. 

In order to assist manufacturers and applicants in conducting safety testing, FDA is recommending that if in vitro mutagenicity testing is contemplated, an enhanced Ames assay be used to assess whether an NDSRI poses a mutagenic risk.  A negative result in a valid enhanced Ames assay may be used to support a higher limit for an NDSRI; however, manufacturers and applicants should note that FDA may request additional safety data, beyond the enhanced Ames assay to support alternative AI limits.  The following recommendations represent FDA’s current thinking; data gaps in the information available to support the safety of NDSRIs remain to be addressed with future research.

The Organisation for Economic Co-operation and Development (OECD)’s Test Guideline No. 471 “Bacterial Reverse Mutation Test” provides standard recommendations for the conduct of the bacterial reverse mutation test (also known as the Ames assay) to assess the mutagenic potential of a test compound.  For N-nitrosamines, enhanced testing conditions for the Ames assay are recommended by FDA due to the reported reduced sensitivity of the assay under standard conditions for some N-nitrosamines such as N-nitroso-dimethylamine (NDMA).  FDA specifically recommends use of an enhanced Ames assay for NDSRIs because very little is known about the sensitivity of the Ames assay to NDSRIs.  Additionally, NDSRIs generally have a wider variety of functional groups present than typically found in low molecular weight N-nitrosamines (such as NDMA) historically studied, and the additional testing conditions described in the enhanced Ames assay have been shown to be helpful in assessing mutagenic risk for NDSRIs.

If a standard Ames assay is conducted on an NDSRI and produces a positive result, FDA recommends there is no need to conduct an additional assay using enhanced testing conditions.

The enhanced Ames assay test conditions presented below are informed by work conducted by FDA’s National Center for Toxicological Research (NCTR) (Li X et al., 2023), as well as other groups, and have been evaluated for a variety of N-nitrosamines, including NDSRIs.  Evaluation of Ames assay test conditions for N-nitrosamines is ongoing with a goal to identify the most robust Ames testing conditions.  The enhanced Ames assay test conditions recommended by FDA for NDSRIs and described below will be updated as warranted.  Deviations from the recommended conditions should be scientifically justified.

Tester strains: S. typhimurium TA98, TA100, TA1535, TA1537, and E. coli WP2 uvrA (pKM101) tester strains should be included.

Type of assay and preincubation time: The pre-incubation method is recommended, and not plate incorporation.  The recommended pre-incubation time is 30 minutes.

Species and concentration of S9:  FDA recommends that Ames assays should be conducted in the absence of a post-mitochondrial fraction (S9), and also in the presence of 30% rat liver S9, as well as 30% hamster liver S9.  FDA also recommends that the rat and hamster post-mitochondrial fractions (S9s) should be prepared from rodents treated with inducers of cytochrome P450 enzymes (e.g., a combination of phenobarbital and β-naphthoflavone).

Negative (solvent/vehicle) control:  FDA recommends that solvents be compatible with the Ames assay as per the OECD 471 guideline.  Solvents can include, but are not limited to:

  • water
  • organic solvents such as acetone, methanol and DMSO

When an organic solvent is used, the lowest possible volume should be included in the pre-incubation mixture with justification to indicate that the volume of solvent does not interfere with metabolic activation of the N-nitrosamine or NDSRI.

Positive controls: Concurrent strain-specific positive controls should be included per the OECD 471 guideline.

Two N-nitrosamines, including NDSRIs, that are known to be mutagenic in the presence of S9 should also be included as positive controls.

The choice of the N-nitrosamine positive controls needs to be justified based on the anticipated metabolism of the N-nitrosamine and the cytochrome P450 enzymes most likely involved.  In addition, if an organic solvent is used to dissolve the test compound, FDA recommends that the volume of organic solvent employed to dissolve the N-nitrosamine positive controls results in a similar concentration as for the test compound in the pre-incubation mix, if possible.

N-Nitrosamine positive controls to consider include:

  1. NDMA (CAS # 62-75-9)
  2. 1-Cyclopentyl-4-nitrosopiperazine (CAS # 61379-66-6)
  3. An NDSRI

All other recommendations for the Ames assay should follow the OECD 471 guideline.


References

OECD Test Guideline No. 471 “Bacterial Reverse Mutation Test” 2020. 

Li X, et al. Revisiting the mutagenicity and genotoxicity of N-nitroso propranolol in bacterial and human in vitro assays. Regul Toxicol Pharmacol. 2023 Jun; 141:105410.

 

 


Revision Table

Updated Information/Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs)

Revision NDSRI Name Change Date Posted*
0 Original Webpage Posting Initial posting of Updated Information/Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs) 8/4/2023**
1 N-nitroso-arformoterol Added to Table 1 10/11/2023
1 N-nitroso-hydrochlorothiazide Added to Table 1 10/11/2023
1 N-nitroso-quinapril Added to Table 1 10/11/2023
1 N-nitroso-nortriptyline Added to Table 1 10/11/2023
1 N-nitroso-desmethyl-imipramine Added to Table 1 10/11/2023
1 N-nitroso-avacopan Added to Table 1 10/11/2023
1 N-nitroso-belumosudil Added to Table 1 10/11/2023
1 N-nitroso-benzonatate Added to Table 1 10/11/2023
1 N-nitroso-deucravacitinib Added to Table 1 10/11/2023
1 N-nitroso-exametazime Added to Table 1 10/11/2023
1 N-nitroso-finerenone Added to Table 1 10/11/2023
1 N-nitroso-leniolisib Added to Table 1 10/11/2023
1 N-nitroso-desmethyl-maralixibat Added to Table 1 10/11/2023
1 N-nitroso-maribavir Added to Table 1 10/11/2023
1 N-nitroso-omidenepag isopropyl Added to Table 1 10/11/2023
1 N-nitroso-pafolacianine Added to Table 1 10/11/2023
1 N-nitroso-viloxazine Added to Table 1 10/11/2023
1 N-nitroso-lorcaserin Deleted from Table 1 10/11/2023
1 N-nitroso-isoxsuprine Deleted from Table 1 10/11/2023
1 N-nitroso-desmethyl-azithromycin Added to Table 1 10/11/2023
1 N-nitroso-fluoxetine Added to Table 2 10/11/2023
2 N-nitroso-ciprofloxacin Added to Table 1 12/1/2023
2 N-nitroso-ciprofloxacin Deleted from Table 2 12/1/2023
2 N-nitroso-atomoxetine Deleted from Table 1 12/1/2023
3 N-nitroso-duloxetine Added to Table 3 2/23/2024
3 N-nitroso-bumetanide Added Testing Method for the Determination of N-nitroso-bumetanide 2/23/2024
3 N-nitroso-propranolol Added Testing Method for the Determination of N-nitroso-propranolol 2/23/2024
3 N-nitroso-varenicline Added Testing Method for the Determination of N-nitroso-varenicline 2/23/2024

*The revisions to the webpage associated with the FDA guidance for industry, Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs) are for immediate implementation as of the date of posting.
**The effective date of the FDA guidance for industry, Recommended Acceptable Intake Limits for Nitrosamine Drug Substance-Related Impurities (NDSRIs) is August 7, 2023, when the Notice of Availability was published in the Federal Register.

 

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