ISO 21766:2021
Tobacco and tobacco products — Determination of tobacco-specific nitrosamines in tobacco products — Method using LC-MS/MS
This document specifies a method for the quantification of four tobacco specific nitrosamines (TSNAs) in tobacco and the following tobacco products: cigarettes, cigars and smokeless tobacco products using reversed phase high performance liquid chromatography with tandem mass spectrometry (LC-MS/MS). The TSNAs determined with this method are: N-nitrosonornicotine (NNN), N-nitrosoanatabine (NAT), N-nitrosoanabasine (NAB) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK).
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Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
5 Reagents
6 Apparatus
7 Preparation
7.1 Preparation of glassware
7.2 Preparation of solutions
7.2.1 Extraction solution, 100 mM ammonium acetate in water
7.2.2 HPLC Mobile Phase A: Water, resistivity ≥ 18,2 MΩ·cm at 25 °C
7.2.3 HPLC Mobile Phase B: 0,1 % acetic acid in methanol
7.3 Preparation of standards
7.3.1 General
7.3.2 Preparation of internal standard solutions
7.3.3 Preparation of calibration standard solutions
8 Sampling
8.1 General
8.2 Sample preparation
8.3 Sample extraction
9 Sample analysis
9.1 General
9.2 Suggested HPLC parameters
9.3 MS/MS parameters
9.3.1 General
9.3.2 Quantification and qualification transitions
9.4 System suitability
9.5 Calibration
9.6 Calculation
10 Repeatability and reproducibility
11 Test report
Annex A (informative) Sample clean-up using solid phase extraction (SPE)
Annex B (informative) Examples of typical chromatograms
Bibliography
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Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
5 Reagents
6 Apparatus
7 Preparation
7.1 Preparation of glassware
7.2 Preparation of solutions
7.2.1 Extraction solution, 100 mM ammonium acetate in water
7.2.2 HPLC Mobile Phase A: Water, resistivity ≥ 18,2 MΩ·cm at 25 °C
7.2.3 HPLC Mobile Phase B: 0,1 % acetic acid in methanol
7.3 Preparation of standards
7.3.1 General
7.3.2 Preparation of internal standard solutions
7.3.3 Preparation of calibration standard solutions
8 Sampling
8.1 General
8.2 Sample preparation
8.3 Sample extraction
9 Sample analysis
9.1 General
9.2 Suggested HPLC parameters
9.3 MS/MS parameters
9.3.1 General
9.3.2 Quantification and qualification transitions
9.4 System suitability
9.5 Calibration
9.6 Calculation
10 Repeatability and reproducibility
11 Test report
Annex A (informative) Sample clean-up using solid phase extraction (SPE)
Annex B (informative) Examples of typical chromatograms
Bibliography