Nitrosamine impurities analysis solution guide

Nitrosamine impurities 

analysis solution guide
 •  Find the right analytical solution for each 

stage of nitrosamine impurity analysis

 •  Confidently detect and quantify genotoxic 

impurities in active pharmaceutical ingredients 
and finished drug products

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Table of contents 

Monitoring genotoxic impurities in pharmaceutical products 

3

Why is nitrosamine impurity analysis in drugs important? 

4

Requirements for nitrosamine analysis 

5

Enabling analytical tools for each stage of nitrosamine impurity analysis 

6

Compliance for all analytical solutions  

7

Absolute confidence with High Resolution Accurate Mass 

8

Cost effective nitrosamine screening with MS/MS 

9

Are you at a cross-roads in choosing your nitrosamine analysis strategy? 

10

Solution summary 

11

LC-HRAM MS solution for absolute mass confirmation 

12 

Learn more about our LC-HRAM MS solution—products and resources  13

GC-HRAM MS solution for nitrosamine analysis gives ultimate  

14 

sensitivity and confidence 

Learn more about our GC-HRAM MS solution—products and resources  15

LC-MS/MS solution for your targeted monitoring needs 

16 

Learn more about our LC-MS/MS solution—products and resources 

17

GC-MS/MS solution for volatile analysis needs with confident targeted approach 18 

Learn more about our GC-MS/MS solution—products and resources 

19

IC Solution for nitrite analysis needs with confident targeted approach 

20 

Learn more about our IC solution—products and resources 

21

Solutions to help address regulatory needs 

22

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Monitoring genotoxic impurities in pharmaceutical products 

Since elevated levels of nitrosamines were detected in certain pharmaceuticals 
as first reported in July 2018, finding the right solutions have become the major 
focus for many pharmaceutical manufacturers. Thermo Fisher Scientific™ as a 
trusted partner for pharmaceutical analysis and quality control offers several 
solutions to fit your needs. We are your partner to:

•  Monitoring the levels of class 1 genotoxic impurities, which are known 

mutagenic carcinogens according to IARC and ICH M7(R1) regulatory 
guidelines

•  Avoid any risks of recalls of your drug products

•  Effortlessly follow and satisfy regulatory guidelines

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Why is nitrosamine impurity analysis in drugs important?

Nitrosamines are small molecular weight chemical substances that are 
probable human carcinogens, and their presence in medicines is considered 
unacceptable by regulators. They are organic compounds featuring a nitroso 
group and amino group. They have the generic formula R

2N-N=O where 

the R groups are typically alkyl in nature, they may be formed during API 
manufacturing—from starting materials, intermediates, reactants, recycling of 
solvents and the presence of nitrites and secondary amines.

In 2018, nitrosamine impurities, including NDMA, were discovered in Valsartan, 
an Angiotensin II receptor blocker (ARB) medication used to treat high blood 
pressure and heart failure. This led to a global recall of Valsartan and several 
other ARB drugs.

First mostly N-Nitrosodimethylamine (NDMA) and N-Nitrosodiethylamine 
(NDEA), two carcinogenic impurities were the target of careful monitoring 
efforts. Since then, several other N-nitrosamines have also been identified 
and are being investigated by regulators: N-Nitrosodiisopropylamine (NDIPA), 
N-Nitrosoethylisopropylamine (NEIPA), N-Nitrosodibutylamine (NDBA), and 
N-Nitroso-N-methyl-4-aminobutyric acid (NMBA).

The outcome that all events has made clear—there’s a recognized need for 
a risk assessment strategy for potential nitrosamines in any pharmaceutical 
product at risk for their presence. 

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Nitrosamine impurity analysis requires robust and sensitive analytical 
methods to ensure confidence in the obtained results. 

The wide Thermo Scientific™ portfolio is proven to be best in class for 
nitrosamine analysis, ensuring your exploratory and routine methods 
are performed as accurately and reliably as possible while maintaining 
requirements from regulatory bodies worldwide. 

Our major products for the analysis of nitrosamines include:
•  Liquid, gas, and ion chromatography for robust separation

•  High resolution, accurate mass (HRAM) mass spectrometry for ultimate 

confidence avoiding false positive results

•  Tandem mass spectrometry, the best tool in routine analysis

•  Single, compliance-ready software solution for all our technology solutions

Performance

Confidence

Service 

and support

Compliance

Process

Product

assessment

Results

Requirements for nitrosamine analysis

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Enabling analytical tools for each stage of  
nitrosamine impurity analysis

 Featured nitrosamine solutions

We’re here for you each step of the way...

GMP compliant 

software for 

all analysis

Routine Screening
• Sensitive, high throughput assays
• Complementary analysis
• LC and GC-MS/MS

• To enable fast and reproducible
• Less hands-on 
• Efficient monitoring 

Highly confident detection
• High resolution accurate mass detection
• Impurity assessment and exploratory analysis
• Confident confirmation
• Routine assay capable

High throughput 

assay automation 

and optimization

Ingredient and product 
monitoring 
• Monitor nitrite and amine 
  levels during manufacturing  
  process and storage
• Risk monitoring

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No matter which solution fit your needs, Thermo Scientific™ Chromeleon™ Chromatography 
Data System (CDS) is going to be there to assure confidence in your compliance. 

Powerful, compliance-ready single solution platform for:

•  Instrument control

•  Data analysis and reporting

•  In a secure, audit-trailed space 

Compliance for all analytical solutions

•  LC-HRAM

•  LC-MS/MS

•  GC-HRAM

•  GC-MS/MS

•  IC

•  Automation

•  Fit within existing laboratory infrastructure of 

chromatographic systems

Stay up-to-date, join  

our community.

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Matrix interferences and or common solvents, such as DMF, from the 
manufacturing process can lead to false-positive results with unit mass 
resolution mass spectrometry. This is especially true if the interfering 
compounds aren’t separated chromatographically. 

We recommend Thermo Scientific™ Orbitrap™ technology as the gold standard 
mass spectrometry technique as a minimum of 45k resolution is critical to 
eliminate false-positive results. Whether you want to perform fast and confident 
nitrosamine impurity screening and or peace-of-mind confirmatory analysis in 
combination with triple quadrupole MS, HRAM MS should be a corner stone in 
your workflow. 

Absolute confidence with High Resolution Accurate Mass

Ultimate confidence—eliminate false positive results with HRAM.

75.0864

75.0650

75.0600

75.0550

75.0492

m/z

3.00

2.00

1.00

0

5.50

5.00

4.00

DMF, 15N isotope

0.4 ppm

NDMA

0.0 ppm

DMF, 13C isotope

0.0 ppm

Mass spectrum of ranitidine drug 
tablet when co-elution of NDMA and 
DMF happens, with a resolution setting 
of 120,000 confident and accurate 
identification of isotope masses 
possible and allows for accurate 
quantitation. 

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Triple quadrupole mass spectrometry is at the forefront of trace analysis 
of nitrosamines in pharmaceuticals. We recommend using this technology 
for screening and in harmony with HRAM for positive result confirmation to 
eliminate false positive results. MS/MS can increase lab productivity and reduce 
workload on your HRAM systems. 

“ Did you know that this strategy is currently used in the same manner for dioxin analysis in 

food and feed? Why? This is an established solution for maximal productivity and ultimate 
confidence in results.”

Cost effective nitrosamine screening with MS/MS

Increase your productivity with the analytical workhorse MS/MS and screen hundreds of samples per day.

Chromatograph of valsartan extract with 
NDMA spiked at 5 ppb (ng/mL) on column

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What are the advantages of each approach?

•  Universal applicability

•  Common availability

•  All FDA nitrosamines are LC amenable

•  Ultimate sensitivity

•  Excellent peak shape

•  Highest column efficiency, best separation

•  Maximum selectivity

•  Zero false positives

•  Ultimate confidence

•  Unknown screening

•  High throughput targeted screening

•  Support of a HRAM confirmatory  

strategy

•  Cost effectiveness and commonality 

MS/MS

HRAM

GC

LC

Are you at a cross-roads in choosing your  
nitrosamine analysis strategy?

THE GOLD STANDARD

Compliance is at the center 

whichever route you choose!

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Solution summary

Nitrosamine analysis

LC-MS

GC-MS

IC

Highly confident identification, quantitation, 
and monitoring

Orbitrap Exploris 120 LC-HRAM

Orbitrap Exploris GC-HRAM

—

Robust, high throughput routine screening

TSQ Quantis LC-MS/MS

TSQ 9000 GC-MS/MS

—

Laboratory workflow automation

TriPlus RSH

TriPlus RSH

—

Single compliance-ready software

Chromeleon CDS Software

Chromeleon CDS Software

Chromeleon CDS Software

Nitrite and nitrate assessment

—

—

HPIC System

Would you like to learn more?  

Contact a specialist now.

Click on each listing for more information 

Featured Application Note

HRAM LC-MS method for the 
determination of nitrosamine 
impurities in drugs

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LC-HRAM MS Solution for absolute mass confirmation

•  Stable chromatography over extended period and hundreds of injections

•  Confidence in analytical results allowing exceptional screening or confirmatory analysis

•  Ease-of-use and compliance-ready software

•  Chromatographic co-elution of NDMA and DMF may 

cause over-estimation (false positive) if mass resolution 
is not sufficient

•  The mass difference between NDMA and DMF 15N 

isotope is only 21 ppm, 0.002 amu difference—requires 
minimum 45K resolution and 3 ppm mass accuracy

75.0864

75.0650

75.0600

75.0550

75.0492

m/z

3.00

2.00

1.00

0

5.50

5.00

4.00

DMF, 15N isotope

0.4 ppm

NDMA

0.0 ppm

DMF, 13C isotope

0.0 ppm

The Thermo Scientific™ Orbitrap Exploris™ 120 mass spectrometer 

system provides high resolution accurate mass assuring absolute 

identification and quantitation of nitrosamines.

Mass spectrum of ranitidine drug tablet when co-elution of NDMA 
and DMF happens, with a resolution setting of 120,000 confident 
and accurate identification of isotope masses possible and allows 
for accurate quantitation. 

Robust separation, 

high load ability

Confident mass 

detection

Fit-for-purpose  

software

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Learn more about our LC-HRAM MS Solution
Products and resources  

Click on each listing for more information 

Would you like to learn more?  

Contact LC-HRAM specialist now.

Robust separation, high load ability 
Impurity separation with (U)HPLC platforms
•  Thermo Scientific™ Acclaim™ PAII Columns

•  Thermo Scientific™ Vanquish™ Flex UHPLC System 

•  Thermo Scientific™ Vanquish™ Core HPLC System 

Confident mass detection 
Mass detection
•  Thermo Scientific™ Orbitrap Exploris™ 120 Mass Spectrometer

•  Thermo Scientific™ Q Exactive™ Plus Mass Spectrometer

Fit-for-purpose software
Software
•  Thermo Scientific™ Chromeleon™ Chromatography Data System 

(CDS) Software

Literature
•  Application Note: HRAM LC-MS method for the determination of 

nitrosamine impurities in drugs

•  Application Note: High-resolution accurate-mass liquid  

chromatography methodology 

•  Article: How to comply with FDA imposed nitrosamine impurity testing

•  Brochure: Vanquish Core

•  Brochure: Orbitrap Exploris 120

•  Brochure: Chromeleon CDS software

•  Brochure: Consumables

Web tools
•  Vanquish Selection tool

•  Nitrosamine impurity analysis

•  Orbitrap LC-MS

•  HPLC and UHPLC platforms

Webinars
•  Nitrosamine analysis; a leading CDMO’s perspective

•  Virtual seminar: Nitrosamine analysis

Products

Resources

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Featured Application Note

A validated method for the 
rapid determination of fifteen 
nitrosamines in metformin drug 
substance

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GC-HRAM MS Solution for nitrosamine analysis 
gives ultimate sensitivity and confidence

•  Ultimate sensitivity

•  Eliminate false positive results

•  Ease-of-use and compliance-ready software

•  Rapid separation of 15 nitrosamines in <12 minutes

•  10× <FDA regulatory limits of 30 ppb (ng/g) <2 ppb

•  Excellent  robustness over 2 weeks continual analysis

Retention time (minutes)

T

IC 

(C

o

u

n

ts

)

5.0e7

4.0e7

3.0e7

2.0e7

1.0e7

0.0e0

4.40

5.00

6.00

7.00

8.00

9.00

10.00

11.00

1.  NDMA
2.  NMEA
3.  NDEA
4.  NIPEA
5.  NDIPA
6. 

NMPA

 7.  NDPA
8.  NEBA
9.  NEPhA
10. NMOR
11. NDBEA
12. NPYR
13. NPIP
14. NDBA
15. NDPhA

12.00

13.00 13.50

1

2

3

4

5

6

7

9

8 10

11, 12

13

14

15

counts

XIC of nitrosamine quantifier ions in a 50 ppb spiked sample. 

The Thermo Scientific Orbitrap Exploris GC-MS system  

provides high resolution accurate mass confirmation  

while beating LOQ requirements. 

Robust separation, 

high load ability

Confident mass 

detection

Fit-for-purpose  

software

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Learn more about our GC-HRAM MS Solution
Products and resources  

Click on each listing for more information 

Robust separation, high load ability 
Impurity separation with GC platforms
•  Thermo Scientific™ TraceGOLD TG-1701 MS Column 

Confident mass detection 
Mass detection
•  Thermo Scientific™ Orbitrap Exploris™ GC Mass Spectrometer

Fit-for-purpose software
Software
•  Thermo Scientific™ Chromeleon™ Chromatography Data System 

(CDS) Software

Literature
•  Applicaton Note: GC Exploris HRAM validation of 15 nitrosamines  

in metformin drug substance

•  Article: How to comply with FDA imposed nitrosamine impurity testing

•  Case Study: GC solutions for nitrosamines

•  Brochure: Orbitrap Exploris GC

•  Brochure: Chromeleon CDS software

•  Brochure: Consumables

Web tools
•  Orbitrap GC-MS

Webinars
•  Nitrosamine analysis; a leading CDMO’s perspective

•  Virtual seminar: Nitrosamine analysis

Products

Resources

Would you like to learn more?  

Contact GC-HRAM specialist now.

Featured Application Note

Highly sensitive and robust  
LC-MS/MS solution for quantitation 
of nitrosamine impurities in 
metformin drug products

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LC-MS/MS Solution for your targeted monitoring needs

•  Stable chromatography over extended period and hundreds of injections

•  Great for screening or confirmatory analysis

•  Ease-of-use and compliance-ready software

•  Vanquish Horizon UHPLC, Hypersil GOLD phenyl column 

coupled to a TSQ Quantis Chromeleon CDS 

•  LOQ 5 ppb using APCI and 10 ppb using HESI 

•  Excellent reproducibility for over 1,000 sample injections

a) Metformin drug sample

NDMA

NMEA

NPYR

NMBA

NDEA

NEIPA

NMPA

NDBA

NDIPA

NDPA

b) 20 ppb spiked sample

The Thermo Scientific Triple Quadrupole systems assure absolute 

confidence in results for any targeted quantitation needs.

XIC of nitrosamine quantifier ions in a 20 ppb spiked sample. APCI 
data are shown.

Robust separation, 

high load ability

Robust and sensitive 

mass detection

Fit-for-purpose  

software

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Learn more about our LC-MS/MS Solution 
Products and resources  

Click on each listing for more information 

Robust separation, high load ability 
Impurity separation with (U)HPLC platforms
•  Thermo Scientific™ Acclaim™ PAII Columns

•  Thermo Scientific™ Vanquish™ Flex UHPLC System 

•  Thermo Scientific™ Vanquish™ Core HPLC System 

Robust and sensitive mass detection 
Mass detection
•  Thermo Scientific™ TSQ Altis Triple Quadrupole  

Mass Spectrometer

•  Thermo Scientific™ TSQ Quantis Triple Quadrupole  

Mass Spectrometer

•  Thermo Scientific™ TSQ Fortis Triple Quadrupole  

Mass Spectrometer

Fit-for-purpose software
Software
•  Thermo Scientific™ Chromeleon™ Chromatography Data System 

(CDS) Software

Literature
•  Application Note: Highly sensitive and robust LC-MS/MS solution for 

quantitation of nitrosamine impurities in metformin drug products

•  Application Note: TSQ Quantis application note

•  Article: How to comply with FDA imposed nitrosamine impurity testing

•  Article: Nitrosamine, a CDMO perspective

•  Brochure: Chromeleon CDS software 

•  Brochure: TSQ Quantis

•  Brochure: Consumables 

•  Brochure: Vanquish Core

Web tools
•  Nitrosamine impurity analysis

•  TSQ LC-MS/MS portfolio

•  HPLC and UHPLC platforms

Webinars
•  Nitrosamine analysis; a leading CDMO’s perspective

•  Virtual seminar: Nitrosamine analysis

Products

Resources

Would you like to learn more?  

Contact LC-MS/MS specialist now.

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GC-MS/MS Solution for volatile analysis needs with 
confident targeted approach

•  Faster data evaluation and increased confidence with exceptional levels of selectivity, 

sensitivity, and linear dynamic range

•  Versatility of full-scan, high-resolution accurate-mass data to screening and quantitation

•  Intuitive instrument control and method templates

•  Results that are compliant with the CFDA and U.S. 

FDA standard methods for nitrosamines detection and 
quantitation in Valsartan

•  Three recommended instrumental approaches  

(GC-MS, HS-GC-MS, GC-MS/MS)

•  The static 

headspace 
injection technique 
offers a simplified 
workflow for 
sample handling

Chromatograph of 
valsartan extract with 
NDMA spiked at  
5 ppb (ng/mL) on column

For high-throughput analytical laboratories, the  

Thermo Scientific TSQ 9000 GC-MS/MS system provides sensitive, 

specific quantitation of target compounds.

High efficiency 

separation

Robust and sensitive 

mass detection

Fit-for-purpose  

software

Featured Application Note

Determination of genotoxic 
nitrosaminesin Valsartan with 
gas chromatography and
mass spectrometry

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Learn more about our GC-MS/MS Solution 
Products and resources  

Click on each listing for more information 

High efficiency separation
Impurity separation with (U)HPLC platforms
•  Thermo Scientific™ TraceGOLD TG-1701 MS Column 

Robust and sensitive mass detection
Mass detection
•  Thermo Scientific™ TSQ 9000 GC-MS/MS System

Fit-for-purpose software
Software
•  Thermo Scientific™ Chromeleon™ Chromatography Data System 

(CDS) Software

Literature
•  Application Note: Determination of genotoxic nitrosamines in 

Valsartan with gas chromatography and mass spectrometry

•  GC Exploris HRAM validation of 15 nitrosamines in metformin drug 

substance

•  Overcoming the challenges of nitrosamine impurities in drugs

•  Brochure: Thermo Scientific TSQ 9000 Triple Quadrupole  

GC-MS/MS System

•  Brochure: Chromeleon CDS software 

•  Brochure: Consumables

Web tools
•  Nitrosamine impurity analysis

•  Triple Quadrupole GC-MS/MS

Webinars
•  Nitrosamine analysis; a leading CDMO’s perspective

•  Virtual Seminar: Nitrosamine analysis

Products

Resources

Would you like to learn more?  

Contact a GC-MS/MS specialist now.

Nitrite in ranitidine drug product (S7) using a Dionex IonPac  
AS19-4μm column

0

5

10

15

20

25

30

Minutes

-2

0

7

1

Peak 

Analyte 

Conc (µg/L)

1 

Nitrite 

95.5

mAU

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IC Solution for nitrite analysis needs with confident 
targeted approach

•  Little or no sample preparation is required

•  Reagent-Free™ (RFIC™) ion chromatography system with electrolytically generated KOH eluent

•  Multi drug product workflow

•  The LOD of nitrite in a pharmaceutical sample is  

0.918 ppm (µg/g API)

•  The method is accurate and precise 

•  Applied to seven pharmaceutical samples, including 

metformin, losartan, ranitidine, and diphenhydramine

The UV absorbance detection allows the sensitive and selective 

detection of nitrite without interference from high amounts of chloride.

High capacity 

columns

Selective  

separations

Fit-for-purpose  

software

Featured Application Note

Determination of nitrite in 
pharmaceuticals

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Learn more about our IC Solution
Products and resources  

Click on each listing for more information 

High capacity columns
Impurity separation with (U)HPLC platforms
•  Thermo Scientific™ Dionex IonPac ™ AS19-4µm Column

Selective separation
Ion chromatography
•  Thermo Scientific™ Dionex™ ICS-6000 Capillary HPIC™ System

Fit-for-purpose software
Software
•  Thermo Scientific™ Chromeleon™ Chromatography Data System 

(CDS) Software

Literature
•  Application Note: Determination of nitrite in pharmaceuticals

•  Brochure: Thermo Scientific Dionex ICS 6000 HPIC System

•  Brochure: Chromeleon CDS software 

•  Brochure: Consumables

Web tools
•  IC home page

•  Nitrosamine impurity analysis

Webinars
•  Nitrosamine analysis; a leading CDMO’s perspective

•  Virtual Seminar: Nitrosamine analysis

Products

Resources

Would you like to learn more?  

Contact a IC specialist now.

For Research Use Only. Not for use in diagnostic procedures. © 2021 Thermo Fisher Scientific Inc. All rights reserved.   
All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. XX74154-EN 1221M

 Find out more at thermofisher.com/nitrosamine

Need more information? Contact a specialist

Solutions to help address regulatory needs

Requirement 

Example approach

API

Separation

Detection

Download

Fit-for-purpose, 
reliable 
quantitative 
screening

Highly sensitive and robust  
LC-MS/MS solution for quantitation 
of nitrosamine impurities in 
metformin drug products

Metformin

LC

Triple quad MS

Determination of genotoxic 
nitrosamines in Valsartan

Valsartan 

GC

Triple quad MS

Future proof your 
lab with absolute 
confidence and 
quantitation

A validated method for the rapid 
determination of fifteen nitrosamines 
in metformin drug substance

Metformin

GC

High resolution 

MS

HRAM LC-MS method for the 
determination of nitrosamine 
impurities in drugs

Ranitidine

LC

High resolution 

MS

HRAM LC-MS methodology for 
the determination and quantitation 
of nitrosamine impurities in drug 
products 

Ranitidine

Valsartan

LC

High resolution 

MS

Overcoming the challenges of 
nitrosamine impurities in drugs:  
What Pharmaceutical QA/QC 
laboratories need to know

Metformin

Valsartan 

GC

High resolution 

MS Triple  

quad MS

Ingredient 
and product 
monitoring

Determination of dimethylamine 
and nitrite in pharmaceuticals by 
ion chromatography to assess the 
likelihood of nitrosamine formation

Multiple

IC

UV / CD

Technical and online support:  
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performance is our goal. Whether you’re looking for an 
instrument manual or spare parts, want to submit a repair 
request, or check on the status of your warranty or service 
contract, we have every support option you’re looking for.
thermofisher.com/technicalresources

The lab-forward trade-in program
A lab-changing experience. Our trade-in process is a highly 
personalized experience that starts with a conversation and 
doesn’t end until your new instrument is successfully up and 
running. We help you assess your lab’s needs and select a 
new system that is the best fit for your lab. We’ll let you 
know the discounted, limited-time new system pricing you’re 
eligible for based on your system trade-in.

If you need help with financing, we have a wide array of 
financing solutions available, but we don’t stop there. Our 
service team will install your new system and arrange for the 
responsible disposal of your current one. Most importantly, 
our personalized support goes well beyond installation. 
 We’ll also help you transfer methods and train your staff. 
Our goal is to ensure, from start to finish, your trade-in is a 
lab-changing experience for the better. 
thermofisher.com/speedofscience

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