A volatilomics analytical protocol employing solid phase microextraction coupled to GC × GC-MS analysis and combined with multivariate chemometrics for the detection of pomegranate juice adulteration

In this work, a solid-phase microextraction (SPME) method combined with two-dimensional gas chromatography coupled to mass spectrometry (GC × GC-MS) was optimized and used to assess the authenticity of pomegranate juice to prevent fraudulent practices. A divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber was used for the extraction of the volatiles. The critical parameters that affect the extraction process, such as the sample volume, and the extraction time were studied. The optimized protocol involved the addition of 15 mL of juice in 50 mL vial and saturation with 30% w/v NaCl.The extraction was carried out within 45 min under 1000 rpm stirring and was applied in the analysis of real juice samples to assess authenticity and detect low levels of pomegranate juice adulteration with grape and apple juice down to 1%. Commercially available pomegranate juice samples were acquired (n1 = 6) and adulterated with 1% of apple juice (n2 = 6), 1% of grape juice (n3 = 6), and a mixture of 1% apple juice and 1% grape juice (n4 = 6). Authentic pomegranate juice samples and adulterated mixtures were analyzed by SPME-GC × GC-MS. The analysis resulted in the identification of 123 volatile compounds that were further processed with chemometric tools. Principal component analysis (PCA) was employed to visualize the clustering of the samples, and a two-way orthogonal partial least squares discriminant analysis (O2PLS-DA) chemometric model was developed and successfully classified the samples to authentic pomegranate juice or adulterated with an explained total variance of 87.4%. The O2PLS-DA prediction model revealed characteristic volatile markers that could be used to detect pomegranate juice fraud.