The potential of enzymatic hydrolysis with HS-SPME-GC×GC-ToFMS for the study of volatile signature of grapes

This works focuses on developing an optimized methodology for analyzing free and bound volatile compounds in grapes using enzymatic hydrolysis with AR2000 enzyme and HS-SPME-GC × GC-ToFMS. To achieve this purpose, a previous optimization step of enzymatic hydrolysis was implemented and optimal conditions for enzymatic hydrolysis were established using 50 mg of AR2000 enzyme at 35 °C for 24 h, while the best extraction conditions were found to be 40 min at 60 °C. Applying this methodology to clones of Moreto grapes from harvest of 2022 and 2023, it was possible to identify and semi-quantify sixty volatile compounds, with more compounds identifiable in 2022 than in 2023. The results also showed the influence of crop year and genetic intravariability. • Analysis of free and bound volatile compounds in one injection. • Enzymatic hydrolysis coupled to HS-SPME - GC x GC-ToFMS. • 60 varietal volatile compounds were identified in Moreto grapes. • Harvest year and genetic variability influence varietal volatile profile of grapes.