Dr. Anoop Kumar - SCIEX, India

SCIEX LC-MS/MS solution for nitrosamine impurities

Anoop Kumar, Ph.D | Business Manager RUO-MKT-11-11459-A © 2020 DH Tech. Dev. Pte. Ltd.

1

Agenda

• SCIEX product portfolio

‒ Nominal mass spectrometry
‒ High resolution mass spectrometry

• Introduction of nitrosamine impurities

‒ Origin and chemical structure of nitrosamine
‒ Potential sources of nitrosamine impurities
‒ Regulatory guidelines and limits

• Analytical method of nitrosamine using LC-MS/MS system

‒ Valsartan, Losartan, Ranitidine

• SCIEX QTRAP® Systems for quantitative and qualitative workflow for nitrosamines
• Global reference LC-MS/MS method and SCIEX technical notes

2 © 2020 DH Tech. Dev. Pte. Ltd.

2020 LC-MS Portfolio

SCIEX Triple Quad™ and QTRAP® systems

3500 Series 4500 Series 5500+ Series 6500+ Series and
SelexION® Technology
Pushing the limits
TripleTOF® 6600+ System
TripleTOF® 5600+ System X500 QTOF System With SelexION® Technology

3 ExionLC™ systems SCIEX HRMS products © 2020 DH Tech. Dev. Pte. Ltd.

SCIEX Triple Quad™ © 2020 DH Tech. Dev. Pte. Ltd.
and QTRAP® 4500 LC-
MS/MS System

TECHNICAL OVERVIEW

4

SCIEX Triple Quad™ and QTRAP® 4500 LC-MS/MS System

QTRAP 4500 SYSTEM Hardware design

Engineered with leading technology 33rrdd qQuuaadrdupcoalne cwaonrk
awsorTkRaAs PLITininQQTTRRAAPP
vveerrssiioonn

Qurved LINAC® Collision Cell

Turbo V™ Ion Source and
Curtain Gas™ Interface

High pressure Q0 and
QJet® Ion Guide

5 © 2020 DH Tech. Dev. Pte. Ltd.

SCIEX Triple Quad™ © 2020 DH Tech. Dev. Pte. Ltd.
5500+ LC-MS/MS
System – QTRAP®
Ready

TECHNICAL OVERVIEW

6

SCIEX Triple Quad™ 5500+ LC-MS/MS System – QTRAP® Ready

• For labs that need to quantify a large list of targets and demand

high quality quantification results in the 1st injection.

• Only the SCIEX 5500+ System delivers high-end speed and

dynamic range, with an upgrade path to include QTRAP®
System functionality

• Fast polarity switching speed (5 msec)
• Extended linear dynamic range (6 orders)
• High energy dynode (HED) detector technology
• Dual QJet® Ion Guide

7 © 2020 DH Tech. Dev. Pte. Ltd.

SCIEX Triple Quad™ 5500+ LC-MS/MS System – QTRAP® Ready

HYBRID TRIPLE QUADRUPOLE LINEAR ION TRAP TECHNOLOGY

New Dual QJet® Ion Guide

8 © 2020 DH Tech. Dev. Pte. Ltd.

QTRAP® Systems © 2020 DH Tech. Dev. Pte. Ltd.

9

Requirements for impurity profiling laboratories SCIEX Triple Quad™ QTRAP®

• Variety of samples in analytical R&D © 2020 DH Tech. Dev. Pte. Ltd.

labs

• LC-MS/MS instruments involved in

the qual and quant data generation

• Need instruments for qualitative

(linear ion trap) and quantitative
(triple quadrupole) applications with
out any compromise in data quality
(very important)

• Two different applications

(qual/quant) area need two different
mass analyzers for best
performance

10

Enhanced resolution data of reserpine using QTRAP® 4500 System

BETTER RESOLUTION AND MASS ACCURACY WITH LINEAR ION TRAP

Exact Mass of Reserpine
(M+H)+: 609.28
Peak width (FWHH): 0.15amu

11 © 2020 DH Tech. Dev. Pte. Ltd.

ER scan - useful to obtain better resolution 4+

3+
2+

12 © 2020 DH Tech. Dev. Pte. Ltd.

Nitrosamines © 2020 DH Tech. Dev. Pte. Ltd.

13

Nitrosamines

INTRODUCTION POTENTIAL ISSUE AND IMPACT

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

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

• Although there are reports of its presence in some • Worldwide review of various sartans and ranitidine

foods and drinking water supplies products has been initiated

• Their presence in medicines is considered • This issue led various recall of several products

unacceptable from USA and EU markets

• Regulators have now strict manufacturing

requirements for these medicines

14 https://www.who.int/medicines/publications/drugalerts/InformationNote_Nitrosamine-impurities/en/ © 2020 DH Tech. Dev. Pte. Ltd.

Origin and chemical structure of nitrosamine 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 © 2020 DH Tech. Dev. Pte. Ltd.

15

Regulatory guideline for nitrosamine impurities

Specification for nitrosamines Analytical data for nitrosamines in sartan products by USFDA
in sartan products

Spec ppm
Level
LOQ 0.03

LOD 0.01
0.005

NDMA in ranitidine products

Ref: Spec ppm
1. USFDA published method Level
LOQ 0.09
https://www.fda.gov/media/125478/download
LOD 0.03
2. European medical agency 0.01
EMA/217823/2019
© 2020 DH Tech. Dev. Pte. Ltd.
(assessment report)

16

SCIEX LC-MS/MS © 2020 DH Tech. Dev. Pte. Ltd.
method

VALSARTAN, LOSARTAN AND
RANITIDINE

17

Valsartan method and results

USFDA/EU limits (ppm) SCIEX method (ppm) QTRAP® 5500 System MS conditions
Specification Limit 0.03 SL 0.03

LOQ 0.01 LOQ 0.01

LOD 0.005 LOD 0.005

Chromatographic profile of six nitrosamine in valsartan
Chromatographic conditions

18 © 2020 DH Tech. Dev. Pte. Ltd.

Calibration curve of six nitrosamines in valsartan API (0.005 – 5.0 ppm)

NDMA NEIPA
NMBA NDIPA
NDEA NDBA

19 © 2020 DH Tech. Dev. Pte. Ltd.

Losartan method and results

USFDA/EU limits (ppm) SCIEX method (ppm) QTRAP® 4500 System MS conditions
Specification Limit 0.03 SL 0.03

LOQ 0.01 LOQ 0.01

LOD 0.005 LOD 0.005

Chromatographic profile of six nitrosamine in losartan

Chromatographic conditions

20 © 2020 DH Tech. Dev. Pte. Ltd.

Calibration curve and statistical data

Representative calibration curves for six nitrosamine compounds Precision and accuracy data of nitrosamines in losartan.
in the losartan assay with r2 value of >0.99. Standard curves were plotted from 0.2 ng/mL to 153.6 ng/mL
for nitrosamine compounds, with N=6 replicates at each level.
21
© 2020 DH Tech. Dev. Pte. Ltd.

Ranitidine - NDMA method and results

USFDA/EU limits (ppm) SCIEX method (ppm) QTRAP® 4500 System MS conditions

SL 0.09 SL 0.09
LOQ 0.03
LOQ 0.03

LOD 0.01 LOD 0.01

Chromatographic profile NDMA in ranitidine

Chromatographic conditions

22 © 2020 DH Tech. Dev. Pte. Ltd.

NDMA in ranitidine - calibration curve and statistical data

NDMA:
75/43

NDMA:
75/58

NDMA: 75/43 NDMA: 75/58

23 © 2020 DH Tech. Dev. Pte. Ltd.

QTRAP® System
workflow for nitrosamine
screening

24 © 2020 DH Tech. Dev. Pte. Ltd.

Information dependent acquisition (IDA) workflow

• IDA helps in the on the fly acquisition of the MQMuRaRMdM™(S)C(IEQXqTrQip)le

full scan spectra (EMS), the isotopic pattern EnEhnanhcaednced
of the masses (ER), the fragmentation reRsoelustoiolnution
pattern (EPI) and can also do the (Q(TTRRAPA®P))
MS/MS/MS
IDA
• This will be an ideal tool for unknown Criteria

sample analysis as this can collect the Enhanced
detailed data from a single LC-MS run Product
Ion Scan
• Further IDA workflows can use the
© 2020 DH Tech. Dev. Pte. Ltd.
combination of scan functions that is
pertaining to the SCIEX Triple Quad™ and
the QTRAP® System thus improving the
power of both qualitative and quantitative
analysis. This is unique to the hybrid
QTRAP instrument

25

IDA workflow on QTRAP® System for non-targeted screening

LIBRARY MATCHING • QTRAP System

helps to perform
non-targeted
screening using
IDA workflow for
compound
confirmation

• Qualitative and

quantitative data
can be generated
in signal injection
workflow

• Improve thoughtput

and confidence in
the of data with
QTRAP Systems

26 © 2020 DH Tech. Dev. Pte. Ltd.

Global reference LC- © 2020 DH Tech. Dev. Pte. Ltd.
MS/MS method

27

Few global references for nitrosamine LC-MS/MS testing

28 © 2020 DH Tech. Dev. Pte. Ltd.

OMCL network https://www.edqm.eu/en/ad-hoc-projects-
omcl-network
29
© 2020 DH Tech. Dev. Pte. Ltd.

SCIEX technical notes on analysis of nitrosamine impurities

• Easy, sensitive and reproducible

method

• Achieving the required LOQ and

LOD as per regulatory
requirements

30 © 2020 DH Tech. Dev. Pte. Ltd.

Acknowledgements EMEAI Pharma Business Unit

India Support Team • Faran Sanchez, Ph.D
• Jinaru Stahl, Ph.D
• Manoj Pillai, Ph.D • Jack Steed
• Aman Sharma • Jeyan Albert
• Sandeep Choudhary, Ph.D • Bhavya Shukla
• Chandrasekhar
• Pankaj, Ph.D © 2020 DH Tech. Dev. Pte. Ltd.

India Marketing Team

• Parikshit Gokhale
• Vallari Saksena

31

Thanks and Questions

32 © 2020 DH Tech. Dev. Pte. Ltd.

[email protected] © 2020 DH Tech. Dev. Pte. Ltd.
[email protected]
Phone: 9958873665

33

Trademarks / Licensing

The SCIEX clinical diagnostic portfolio is For In Vitro Diagnostic Use. Rx Only.
Product(s) not available in all countries. For information on availability, please
contact your local sales representative or refer to https://sciex.com/diagnostics.
All other products are For Research Use Only. Not for use in Diagnostic
Procedures.
Trademarks and/or registered trademarks mentioned herein are the property of
AB Sciex Pte. Ltd. or their respective owners in the United States and/or certain
other countries.
© 2020 DH Tech. Dev. Pte. Ltd. RUO-MKT-11-11459-A

34 © 2020 DH Tech. Dev. Pte. Ltd.