Thorough investigation of the phenolic profile of reputable Greek honey varieties:varietal discrimination and floral markers identification using liquid chromatography–high-resolution mass spectrometry

Honey is a highly consumed commodity due to its potential health benefits upon certain consumption, resulting in a high market price. This fact indicates the need to protect honey from fraudulent acts by delivering comprehensive analytical methodologies. In this study, targeted, suspect and non-targeted metabolomic workflows were applied to identify botanical origin markers of Greek honey. Blossom honey samples (n = 62) and the unifloral fir (n = 10), oak (n = 24), pine (n = 39) and thyme (n = 34) honeys were analyzed using an ultra-high-performance liquid chromatography hybrid quadrupole time-of-flight mass spectrometry (UHPLC-q-TOF-MS) system. Several potential authenticity markers were revealed from the application of different metabolomic workflows. In detail, based on quantitative targeted analysis, three blossom honey markers were found, namely, galangin, pinocembrin and chrysin, while gallic acid concentration was found to be significantly higher in oak honey. Using suspect screening workflow, 12 additional bioactive compounds were identified and semi-quantified, achieving comprehensive metabolomic honey characterization. Lastly, by combining non-targeted screening with advanced chemometrics, it was possible to discriminate thyme from blossom honey and develop binary discriminatory models with high predictive power. In conclusion, a holistic approach to assessing the botanical origin of Greek honey is presented, highlighting the complementarity of the three applied metabolomic approaches

Honey Phenolic Compound Profiling and Authenticity Assessment Using HRMS Targeted and Untargeted Metabolomics

Honey consumption is attributed to potentially advantageous effects on human health due to its antioxidant capacity as well as anti-inflammatory and antimicrobial activity, which are mainly related to phenolic compound content. Phenolic compounds are secondary metabolites of plants, and their content in honey is primarily affected by the botanical and geographical origin. In this study, a high-resolution mass spectrometry (HRMS) method was applied to determine the phenolic profile of various honey matrices and investigate authenticity markers. A fruitful sample set was collected, including honey from 10 different botanical sources (n = 51) originating from Greece and Poland. Generic liquid–liquid extraction using ethyl acetate as the extractant was used to apply targeted and non-targeted workflows simultaneously. The method was fully validated according to the Eurachem guidelines, and it demonstrated high accuracy, precision, and sensitivity resulting in the detection of 11 target analytes in the samples. Suspect screening identified 16 bioactive compounds in at least one sample, with abscisic acid isomers being the most abundant in arbutus honey. Importantly, 10 markers related to honey geographical origin were revealed through non-targeted screening and the application of advanced chemometric tools. In conclusion, authenticity markers and discrimination patterns were emerged using targeted and non-targeted workflows, indicating the impact of this study on food authenticity and metabolomic fields

ESR Dating Ungulate Teeth and Molluscs from the Paleolithic Site Marathousa 1, Megalopolis Basin, Greece

At 37°24′ N 22°8′ E, the Megalopolis Basin lies in the central Peloponnese Peninsula, southwestern Greece. In the Megalopolis Basin at ~350 m amsl, the Paleolithic site, Marathousa 1, sits within a palustrine/lacustrine clastic package between Lignite Seams III and II, that both likely correlate with interglacial periods. At Marathousa 1, immediately below Lignite Seam III, lies a clayey-silty sand layer with a horizon rich in molluscs ranging from ~20–40 cm thick. About 0.8–1.3 m below the shell-rich horizon (SRH), lacustrine silty to muddy sands rich in organic matter yielded Paleolithic lithic artefacts associated with Middle Pleistocene fauna, some with cut marks and possible bone knapping, found within palustrine/lacustrine clastic deposits. Since ESR (electron spin resonance) can date teeth and molluscs aged >2 Ma, two bivalve samples, AM66 and AM65, five subsamples from a cervid molar, AT39, and one subsample from another cervid molar, AT68, were independently dated by ESR from Marathousa 1. To calculate the ages, time-averaged cosmic and time- and volumetrically-averaged sedimentary dose rates were calculated using past water depths and sedimentation rates as determined from paleontological and geological criteria. Found in the SRH in Layer UA2, AM66 and AM65 averaged 488 ± 37 ka, which correlates with MIS 13a. Because the bivalves sat stratigraphically above the artefacts and mammalian fossils, their ages constrain the ESR ages for the teeth deposited below. Lying on the unconformity at the base of Layer UA3c with UA4, and its correlative unconformity at the Layer UB4c/UB5 boundary, sat the dated teeth from large mammals. Because the bones in the Palaeoloxodon antiquus skeleton lay in quasi-anatomical association, the likelihood for fossil reworking on the Layer UB3c/UB4 surface is low. Isochron analysis suggests that using a U uptake model with p = 2 provides the most accurate ages for AT39. With p = 2, AT39 dates to 503 ± 13 ka, while AT68 dates to 512 ± 34 ka. Nonetheless, two to three more teeth and molluscs should be dated to confirm these ages, when more samples suitable for ESR dating are found. Both tooth ages correlate well with early MIS 13, an interglacial period with cooler mean global temperatures compared to MIS 11 or 9. Assuming that the archaeological site formed in one event, rather than as a palimpsest, the data suggest that hominins processed elephant and other faunal carcasses along the shores of a shallow lake or marsh in the Megalopolis Basin at 503 ± 12 ka. Between the two horizons dated here, their sedimentation rate averaged 4.8 ± 1.8 to 7.8 ± 2.9 cm/ka