Determination of N-nitrosamines in processed meats by liquid extraction combined with gas chromatography-methanol chemical ionisation/mass spectrometry

Abstract

A simple, accessible and reproducible method was developed and validated as an alternative for the determination of nine volatile N-nitrosamines (NAs) in meat products, using a low volume of organic solvent and without requiring specific apparatus, offering the possibility of practical implementation in routine laboratories. The NAs were extracted with dichloromethane followed by a clean-up with phosphate buffer solution (pH 7.0). The extracts were analysed by gas chromatography-chemical ionisation/mass spectrometry (GC-CI/MS) in positive-ion mode using methanol as reagent. Limits of detection and quantification, recovery and reproducibility were determined for all NAs (N-nitrosodimethylamine, N-nitrosomethylethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosodipropylamine, N-nitrosomorpholine, N-nitrosopiperidine, N-nitrosodibutylamine and N-nitrosodiphenylamine). Satisfactory sensitivity and selectivity were obtained even without concentrating the extract by solvent evaporation, avoiding the loss of the nine NAs studied. Limits of detection ranged from 0.15 to 0.37 µg kg⁻¹, whereas limits of quantification ranged from 0.50 to 1.24 µg kg⁻¹. Recoveries calculated in cooked ham that had been spiked at 10 and 100 µg kg⁻¹ were found to be between 70% and 114% with an average relative standard deviation of 13.2%. The method was successfully used to analyse five samples of processed meat products on the day of purchase and 7 days later (after storage at 4°C). The most abundant NAs found in the analysed products were N-nitrosodipropylamine and N-nitrosopiperidine, which ranged from 1.75 to 34.75 µg kg⁻¹ and from 1.50 to 4.26 µg kg⁻¹, respectively. In general, an increase in the level of NAs was observed after the storage period. The proposed method may therefore be a useful tool for food safety control once it allows assessing the profile and the dietary intake of NAs in food over time.

Keywords:

• N-nitrosamines
• meat products
• liquid extraction
• methanol chemical ionisation
• gas chromatography/mass spectrometry

Acknowledgements

The authors wish to thank D. Bélanger for providing technical assistance with the GC-MS equipment. The authors are also grateful to Agriculture and Agri-Food Canada (Food Research and Development Centre) for access to the infrastructure required for this study.

Additional information

Funding

The authors are grateful to the Brazilian Federal Agency for the Support and Evaluation of Graduate Education (CAPES) for providing financial contributions [process number 6116-13-2]; and to Agriculture and Agri-Food Canada (Food Research and Development Centre) for providing funds required for this study.