ISO 19290:2021 Cigarettes — Determination of tobacco specific nitrosamines in mainstream cigarette smoke — Method using LC-MS/MS

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Cigarettes — Determination of tobacco specific nitrosamines in mainstream cigarette smoke — Method using LC-MS/MS是国际标准化组织于2021-02发布的ISO标准，适用于国际范围。

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INTERNATIONAL STANDARD ISO 19290:2021(E)

Cigarettes — Determination of tobacco specific nitrosamines in mainstream cigarette smoke — Method using LC-MS/MS

WARNING — The use of this document can involve hazardous materials, operations and equipment. This document does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this document to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

1 Scope

This document specifies a method for the quantification of four tobacco specific nitrosamines (TSNAs) in the total particulate matter of mainstream cigarette smoke by using reversed phase high performance liquid chromatography with tandem mass spectrometry (LC-MS/MS). The quantified TSNAs are: N-nitrosonornicotine (NNN), N-nitrosoanatabine (NAT), N-nitrosoanabasine (NAB) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK).

2 Normative references

The following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.

ISO 3308, Routine analytical cigarette-smoking machine — Definitions and standard conditions

ISO 3402, Tobacco and tobacco products — Atmosphere for conditioning and testing

ISO 4387, Cigarettes  —  Determination  of  total  and  nicotine-free  dry  particulate  matter  using  a  routine 

analytical smoking machine

ISO 8243, Cigarettes — Sampling

3 Terms and definitions

For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at https://w ww. iso .org/ obp

— IEC Electropedia: available at http://w ww.e lectropedia. org/ 3.1

tobacco specific nitrosamines

TSNAs

four nitrosamines found predominantly in tobacco: N-nitrosonornicotine (NNN), N-nitrosoanatabine (NAT), N-nitrosoanabasine (NAB) and 4-(N-Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)

[SOURCE: ISO 22303:2008, 3.1]

ISO 19290:2021(E)

4 Principle

Cigarettes are smoked on a standard smoking machine. The mainstream smoke is trapped on a glass-

fibre filter pad. After addition of an internal standard, the total particulate matter collected on the glass-fibre filter pad is extracted with 100 mM ammonium acetate solution using a shaker.

The extract is syringe filtered through a 0,45 μm PTFE column directly into an auto sampler vial.

The samples are subjected to reversed phase high performance liquid chromatography (HPLC) and quantified via tandem mass spectrometry (MS/MS).

5 Apparatus

Usual laboratory apparatus and equipment for use in preparation of samples and standards and in particular the following items. All glassware shall be cleaned before use to avoid any contamination.

5.1 Equipment needed to perform conditioning of cigarettes, in accordance with ISO 3402.

5.2 Equipment needed to perform marking for butt length of cigarettes.

5.3 Equipment needed to perform smoking of cigarettes, in accordance with ISO 3308.

5.4 Analytical balance, capable of measuring to at least four decimal places.

5.5 Centrifuge tubes, 50 ml.

5.6 Dispenser, of capacity 20 ml for extraction solution.

5.7 Gas-tight syringes, of capacity 250 μl.

5.8 Automated volumetric pipette.

5.9 Shaker.

5.10 High performance liquid chromatograph coupled to tandem mass spectrometer (LC-MS/

MS), consisting of:

5.10.1 Binary pump.

5.10.2 Autosampler.

5.10.3 Tandem mass spectrometer.

5.10.4 Data collection system.

®1)

5.10.5 LC column: XTerra MS C18 , 2,5 μm, 2,1 mm × 50 mm or equivalent.

6 Reagents

Use only reagents of recognized analytical reagent grade.

1) XTerra MS C18 is an example of a suitable product available commercially. This information is given for the convenience of users of this document and does not constitute an endorsement by ISO of this product.

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