Overview
ISO 19290:2021 specifies an analytical method for the quantification of four tobacco specific nitrosamines (TSNAs) in the total particulate matter (TPM) of mainstream cigarette smoke using reversed‑phase high performance liquid chromatography with tandem mass spectrometry (LC‑MS/MS). The target analytes are NNN, NAT, NAB and NNK. The standard describes sampling, sample preparation, instrumentation and calculation procedures to deliver robust, reproducible TSNA measurements for product testing and research.
Key Topics and Requirements
- Analytes quantified: N‑nitrosonornicotine (NNN), N‑nitrosoanatabine (NAT), N‑nitrosoanabasine (NAB), 4-(methylnitrosamino)-1-(3‑pyridyl)-1‑butanone (NNK).
- Sample collection: Mainstream smoke is collected on a glass‑fibre (Cambridge) filter pad; total particulate matter (TPM) is used for analysis.
- Internal standards: Use of deuterated analogues (NNN‑d4, NAT‑d4, NAB‑d4, NNK‑d4) for accurate quantification.
- Extraction and cleanup: TPM is extracted (e.g., with 100 mM ammonium acetate), filtered (0.45 µm PTFE) and prepared for LC‑MS/MS analysis.
- Chromatography and detection: Reversed‑phase HPLC separation followed by tandem mass spectrometry (MS/MS) for selective, sensitive detection. Example column type and system components are described (binary pump, autosampler, MS/MS).
- Calibration and calculation: Procedures for preparing calibration standards, constructing calibration curves, calculating TSNA concentrations and reporting mainstream smoke deliveries.
- Quality and reproducibility: Guidance on repeatability, reproducibility and statistical evaluation from interlaboratory experiments.
- Controlled conditions: Cigarette conditioning and machine smoking per ISO 3402 and ISO 3308; emphasis on clean glassware and validated procedures.
- Safety note: The document warns about hazardous materials and the need to follow appropriate laboratory safety and regulatory requirements.
Applications and Who Uses It
- Tobacco industry analytical and R&D laboratories performing product characterization or formulation changes.
- Regulatory authorities and compliance labs measuring TSNA emissions for product surveillance.
- Independent analytical contract labs offering LC‑MS/MS testing services.
- Public health researchers studying toxicant yields and trends in cigarette mainstream smoke.
- Quality control groups seeking standardized, comparable TSNA data across products and studies.
This method supports accurate, reproducible measurement of TSNAs in mainstream cigarette smoke and provides a recognized international protocol for LC‑MS/MS based analysis.
Related Standards
- ISO 3308 - Routine analytical cigarette‑smoking machine (smoking regime)
- ISO 3402 - Conditioning and atmosphere for tobacco testing
- ISO 4387 - Determination of total and nicotine‑free dry particulate matter
- ISO 8243 - Cigarette sampling
Keywords: ISO 19290:2021, TSNAs, LC‑MS/MS, mainstream cigarette smoke, NNN, NNK, NAB, NAT, HPLC‑MS/MS, Cambridge filter pad, tobacco specific nitrosamines.