Untargeted profiling and differentiation of geographical variants of wine samples using headspace solid-phase microextraction flow-modulated comprehensive two-dimensional gas chromatography with the support of tile-based Fisher ratio analysis

The volatile fraction of food, also called the food volatilome, is increasingly used to develop new fingerprinting approaches. The characterization of the food volatilome is important to achieve desired flavor profiles in food production processes, or to differentiate different products, with winemaking being one popular area of interest. In the present research, headspace solid-phase microextraction (HS SPME) coupled to flow-modulated comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (FM GC×GC-TOFMS) was used to characterize geographical-based differences in the volatilome of five white “Grillo” wines (of Sicilian origin), comprising the five sample classes. All wines were produced with the same vinification method in 2019. To minimize the influence of minor bottle-to-bottle differences, three bottles of the same wine were randomly selected, and three samples were collected per bottle, resulting in nine sample replicates per wine. Particular emphasis was devoted to the operational conditions of a novel low duty cycle flow modulator. A fast FM GC×GC-TOFMS method with a modulation period of 700 ms and a re-injection period of 80 ms was developed. Following, the instrumental software was exploited to identify class-distinguishing analytes in the dataset via tile-based Fisher ratio analysis (i.e., ChromaTOF Tile). A tile size of 10 modulations (7 s) on the first dimension and 45 spectra (300 ms) on the second dimension was used to encompass average peak widths and to account for minor retention time shifting. Off-line software was used to apply an ANOVA test. A p-value of 0.01 was applied in order to select the most important class-distinguishing analytes, which were input to principal component analysis (PCA). The PCA scores plot showed distinct clustering of the wines according to geographical origin, although the loadings revealed that only a few analytes were necessary to differentiate the wines. However, a comprehensive flavor profile assessment underscored the importance of all the information output by the ChromaTOF Tile software.