Unveiling the unknow aroma potential of Port wine fortification spirit taking advantage of the comprehensive two-dimensional gas chromatography

Abstract

Port wine is a fortified wine exclusively produced in the Douro Appellation (Portugal) under very specific conditions resulting from natural and human factors. Its intrinsic aroma characteristics are modulated upon a network of factors, such as the terroir, varieties and winemaking procedures that include a wide set of steps, namely the fortification with grape spirit (ca. 77% v/v ethanol). The fortification spirit comprises roughly one fifth of the total volume of this fortified wine, thus it is a potential contributor to the global quality of this beverage, including the aroma notes. Nonetheless, the information about the influence of the fortification spirit on the final aroma of Port wine, as well as the grape spirit volatile composition are extremely limited. Therefore, the main objective of this research is the optimization of an adequate methodology for the in-depth characterization of the fortification grape spirit volatile components, based on the use of advanced gas chromatography (GC×GC-ToFMS), combined with a solvent free solid phase microextraction technique (SPME). To fulfil this objective, the SPME experimental parameters (fiber coating, extraction temperature, and time, sample volume and dilution conditions) were optimized. Also, different column sets (first × second dimensions) were also tested to obtain the best chromatographic resolution and peak capacity. Firstly, the GC×GC-ToFMS experimental parameters were implemented, and the reversed phase column set (polar 1D × nonpolar 2D), with the same diameters in primary and secondary columns, presented advantages compared to the conventional column set (nonpolar 1D × polar 2D) regarding the analytes´ separation. Secondly, the SPME conditions that promoted the highest extraction efficiency were selected: 2.0 mL of spirit (diluted at 10% v/v ethanol) were extract with poly(dimethylsiloxane)/divinylbenzene fiber, at 40˚C, using 10 min of pre-equilibrium followed by 30 min of extraction. An exploratory application was performed using a set of grape spirits, which allowed the detection of hundreds of volatiles, from which 120 were putatively identified. This study adds further insights unveiling the complex nature of the grape spirits chemical volatile data, through the identification of compounds not yet determined in these matrices, some of which are associated with aroma notes highly valued in fortified wines. In addition, these volatile patterns seem to be useful to the spirits distinction/typing.

Acknowledgments:

This work was funded under the AD4PurePort – New range of Port wines, based on an innovative method of selecting fortification spirits), project 39956 – POCI-01-0247-FEDER-039956, supported by the COMPETE 2020 Operational Programme under the PORTUGAL 2020. Thanks are also due to FCT/MEC for the financial support LAQV-REQUIMTE (UIDB/50006/2020) through national funds and co-financed by the FEDER, with a PT2020 Partnership Agreement.