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Chestnut (Castanea sativa) shells (CSS) are a source of bioactive compounds with well demonstrated in-vitro antioxidant properties. Nevertheless, no in-vivo studies have already evaluated this effect. This study evaluated the effects of the oral daily administration of an eco-friendly CSS extract (50 and 100 mg/kg per body weight (b. w.)) to rats regarding in-vivo antioxidant activity, glucose and lipids levels, and metabolomic profiling of poly- phenols by LC-ESI-LTQ-Orbitrap-MS. The results demonstrated the in-vivo antioxidant properties in the animals liver, kidney and blood serum, as well as protective effects against hemolysis and rising of blood glucose and lipids levels. New insights on metabolomic profiling of polyphenols proved their absorption and further biotransformation by phase I (hydrogenation and hydroxylation) and II reactions (glucuronidation, methylation and sulfation). This is the first study that attempted to validate a novel nutraceutical ingredient extracted from CSS by in-vivo assays, corroborating the outcomes screened by in-vitro assays, The authors’ kindly thanks to Sortegel (Sortes, Portugal) for the samples. This work received financial support from national funds (UIDB/50006/2020), project PTDC/ASP-AGR/29277/2017 - Castanea sativa shells as a new source of active ingredients for Functional Food and Cosmetic applications: a sustainable approach, and project 5537 DRI, Sérvia 2020/21 from Portuguese-Serbia Bilateral Cooperation - Development of functional foods incorporating a chestnut shells extract obtained by subcritical water, supported by national funds by FCT / MCTES and co-supported by Fundo Europeu de Desenvolvimento Regional (FEDER) throughout COMPETE 2020 - Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-029277). Diana Pinto (SFRH/BD/144534/2019) and Soraia Pinto (SFRH/BD/144719/2019) are thankful for their PhD grants financed by FCT/MCTES and POPH-QREN and supported by funds from European Union (EU) and Fundo Social Europeu (FSE) through Programa Operacional Regional Norte. Francisca Rodrigues is grateful for her contract (CEECIND/01886/2020) financed by FCT/MCTES—CEEC Individual 2020 Program Contract. Anna Vallverdú-Queralt thanks the Spanish Ministerio de Ciencia, Innovación y Universidades for the Ramon y Cajal contract (RYC-2016-19355).

In this study, atmospheric pressure chemical ionisation-mass spectrometry (APCI-MS) was successfully applied to understand the effect of phenolic compounds on the release of olive oil aroma compounds. Eight aroma compounds were monitored under in-vivo and in-vitro dynamic conditions in olive oil with and without the addition of virgin olive oil (VOO) biophenols. Three model olive oils (MOOs) were set up with identical volatile compounds concentrations using a refined olive oil (ROO). Phenolics were extracted from VOOs and were added to two MOOs in order to obtain two different concentrations of phenolic compounds (P+ = 354 mg kg-1; P++ = 593 mg kg-1). Another MOO was without VOO biophenols (P-). Phenolic compounds impacted both the intensity and time of aroma release. In the in-vivo study, 1-penten-3-one, trans-2-hexenal and esters had lower release in the presence of higher levels of biophenols after swallowing. In contrast, linalool and 1-hexanol had a greater release. The more hydrophobic compounds had a longer persistence in the breath than the hydrophilic compounds. VOO phenolics-proline-rich proteins complexes could explain the binding of aroma compounds and consequently their decrease during analysis and during organoleptic assessment of olive oil.