Quantification of benzoxazinoids and their metabolites in Nordic breads

- Common breads in the Nordic countries contain an array of benzoxazinoids.
- Deuterated internal standards for HBOA and 2-HHPAA were synthesized.
- Method developed for quantification of benzoxazinoids using UPLC-MS/MS.

Benzoxazinoids (Bx) and their metabolites are molecules with suggested health effects in humans, found in cereal grains and consequently in cereal foods. However, to date little is known about the amount of Bx in our diet. In this study, deuterated standards 2-hydroxy-1,4-benzoxazin-3-one (HBOA-d4) and 2-hydroxy-N-(2-hydroxyphenyl) acetamide (HHPAA-d4) were synthesized, to allow quantification of nine Bx and their metabolites in 30 breads and flours from Nordic countries by UHPLC-MS/MS. Samples containing rye had larger amounts of Bx (143-3560 µg/g DM) than the ones containing wheat (11-449 µg/g DM). More Bx were found in whole grain wheat (57-449 µg/g DM) compared to refined wheat (11-92 µg/g DM) breads. Finnish sourdough rye breads were notably high in their 2-hydroxy-N-(2-hydroxyphenyl) acetamide (HHPAA) concentration (40-48 µg/g DM). This new information on Bx content in flours and breads available in the Nordic countries will be useful for future work on determining dietary exposure to Bx.