This study investigated the fermentability and prebiotic potential of ulvan, a sulfated polysaccharide from Ulva lactuca L., using an in vitro human colonic fermentation model. Targeted metabolomics quantified SCFA and tryptophan-derived metabolites to assess saccharolytic activity and fiber-driven modulation of amino acid metabolism.Ulvan supplementation significantly stimulated SCFA production, particularly acetic acid, achieving levels comparable to the well-established prebiotic inulin. In parallel, ulvan selectively modulated tryptophan metabolism, enhancing the production of health-associated metabolites such as indole-3-propionic acid and indole-3-acetic acid, suggesting cross-feeding interactions among saccharolytic and indole-producing taxa. Untargeted metabolomics revealed distinct metabolites associated with ulvan treatment, including cyclic proline dipeptides, which points to a selective stimulation of beneficial microbial species.Collectively, these findings demonstrate that ulvan is a soluble and fermentable dietary fiber, supporting its potential as a functional food ingredient or dietary supplement for promoting gut health.

Virgin olive oil supply chain generates large amounts of pruning residues such as olive leaves, rich in bioactive phenols, but still largely underutilized. Phenols are susceptible to enzymatic degradation, thus proper post-harvest management, especially drying conditions, is crucial to maximizing phenolic and oleuropein content. This study assessed the impact of different drying methods on leaves' phenolic content, establishing a protocol to achieve oleuropein levels > 5% on dry weight, making them suitable for the foods and nutraceuticals industry. The effects of cultivar, harvest season, and storage time before drying were also evaluated by HPLC-DAD-MS. Oven drying at 105 degrees C was identified as optimal, providing oleuropein concentrations even greater than 10% DW. Other approaches like lyophilization, drying at 50 degrees C, and extraction from fresh leaves caused up to 93% oleuropein loss. Delayed drying after harvesting led to oleuropein degradation as well. The study contributed to clarify conflicting results in the literature regarding the impact of different drying methods and temperatures. It also provided suitable protocols for preserving oleuropein in olive leaves and obtaining dried material with oleuropein content greater than 5% w/w, summarized as follows: olive leaves of specific cultivars (e.g., Frantoio) harvested in late winter/spring must be oven-dried at 105 degrees C within three days after harvesting.

Pomegranate peel, accounting for 35-50% of the fruit's weight, is an underutilized by-product rich in ellagitannins and pectin. Although pomegranate juice has already been subjected to fermentation, this study aimed to demonstrate for the first time, that a simple fermentation process of pomegranate peel using Saccharomyces cerevisiae can yield tannin-rich extracts and enhance the potential prebiotic properties of pomegranate polysaccharides. Peels of ‘Wonderful’ and ‘G1’ cultivars, significantly differing in peel thickness, were used for fermentation applying three processes at room temperature monitored at 48 and 72 h. HPLC-DAD analysis showed that yeast-driven fermentation increased total tannin (up to 70% in dry extracts) compared with spontaneous fermentation (40-43%). A potential enhancement of prebiotic activity was associated with the reduction in the molecular weight of native polysaccharides (from a maximum of 705 kDa down to 26 kDa), as evaluated by DLS and DOSY-1H-NMR. Polysaccharides after fermentation were more effectively utilized by Bifidobacterium breve and Lactobacillus plantarum than those from control samples, as demonstrated by in vitro assays. These findings highlight a promising strategy for the valorization of pomegranate peel through the production of functional ingredients highly enriched in tannins and containing potentially prebiotic polysaccharides.

This study compares three headspace-based extraction techniques-Regular solid-phase microextraction (HSSPME), vacuum-assisted SPME (Vac-HS-SPME), and probe-type sorptive extraction (HiSorb)-for the analysis of volatile organic compounds (VOCs) in virgin olive oil (VOO). Using a combined untargeted-targeted GC & times; GC-MS approach, 34 VOCs were selected. Vac-HS-SPME enhanced recovery of semi-volatile compounds linked to oxidative and microbiological defects, underrepresented by HS-SPME and HiSorb. Conversely, HSSPME and HiSorb preferentially extracted low-molecular-weight lipoxygenase (LOX)-derived volatiles associated with green and fruity notes, with HiSorb showing broader coverage and stronger alcohol and aldehyde responses. ANOVA and Tukey's post hoc test (p < 0.05) indicated significant differences among techniques for 24 analytes. These findings provide method-oriented insights into VOC-based workflows for olive oil profiling and suggest the potential applicability of targeted headspace applications in quality control, classification, and sensory characterization of VOO.

Pomegranate peel, accounting for 35–50% of the fruit weight, is an underutilized agri-food by-product. This study applied, for the first time, fermentation with Saccharomyces cerevisiae as a simple and sustainable strategy to simultaneously obtain tannin-rich extracts and polysaccharide fractions with potential prebiotic activity. Peels from two cultivars, Wonderful and G1, differing in peel thickness, were subjected to three fermentation protocols (air- and not air-exposed) and monitored at 25 °C over 48 and 72 h. HPLC-DAD analysis showed that yeast-inoculated fermentation increased total tannin concentration in dry extracts (up to 70%) without inducing chemical modifications to tannin profiles. As determined by Dynamic Light Scattering, fermentation promoted significant depolymerization of native polysaccharides, while DOSY-1H-NMR analyses revealed the presence of reduced molecular weight fractions down to 26 kDa. In vitro growth assays confirmed that fermented polysaccharides were more efficiently utilized as a carbon source by Bifidobacterium breve and Lactiplantibacillus plantarum compared to non-fermented controls, likely thanks to polysaccharide depolymerization induced by fermentation. The study demonstrated that air-exposed S. cerevisiae fermentation was an effective process alternative to chemical or enzymatic hydrolysis for modifying pomegranate peel pectin directly within a complex matrix, while simultaneously enhancing tannin recovery. This approach represents a possible sustainable strategy for pomegranate peel valorization into functional ingredients.

This study suggests original holistic approach based on comprehensive analysis of phenolic compounds (HPLC-DAD), volatile (including pentene dimers) and terpene compounds (HS-SPME-GC-MS) and sensory analysis by the Panel Test to propose practical tools for clustering and quality differentiation of monovarietal extra virgin olive oils (MEVOO). The sample-set consisted of 356 MEVOO samples from 13 major Italian cultivars analyzed across five production years. Hierarchical Cluster Analysis following genetic algorithm-linear discriminant analysis and random forest methods allowed identifying three robust cultivar clusters with similar sensory features and selecting nine significant sensory attributes. Models for samples classification in these clusters were set and externally validated. Terpenes exhibited the best relationships with sensory clustering (reaching approx. 92% accuracy), but combining terpene, volatile and phenolic compounds improved accuracy up to 98%. Finally, two simplified chemometric models based either on 8 terpenes or 12 combined terpene, volatile, and phenolic compounds were proposed for predicting MEVOOs classification in the three clusters.

This study presents a comprehensive chemical and volatilomic characterization of oils exhaustively extracted from blueberry (Vaccinium corymbosum L.) and raspberry (Rubus idaeus L.) pomace. Both oils are dominated by polyunsaturated fatty acids (PUFAs): linoleic acid (36.5 % in blueberry, 49.3 % in raspberry) and alpha-linolenic acid (32.7 % and 28.2 %, respectively), with favorable omega-6/omega-3 ratios of 1.75 (blueberry) and 1.12 (raspberry). Key unsaponifiable bioactives were quantified: raspberry pomace oil (RPO) contained 3620 mg/kg total tocopherols and 4244 mg/kg phytosterols; blueberry pomace oil (BPO) was notable for 509 mg/kg squalene. Polar phenolics (21-32 mg/kg) included p-coumaric and ferulic acids, vanillin, and flavonoids such as naringenin, enhancing antioxidant potential. Headspace SPME-GC-MS quantified 41 volatile organic compounds; BPO's aroma was dominated by terpenes and aldehydes, whereas RPO showed a richer carboxylic acid profile. The use of an indepth analytical approach enabled a thorough assessment of the oils' compositional potential, offering a valuable reference for future applications. These findings provide a foundation for the optimization and improvement of extraction processes aimed at producing sustainable and bioactive-rich ingredients, aligning with circular economy principles to reduce waste and develop high-value products with health and sensory benefits.

Date palm (Phoenix dactylifera L.) by-products from bioethanol production represent an underutilized resource rich in bioactive molecules. This study aims to their valorization through characterization of polysaccharides and phenolic compounds from the Medjool variety, both before and after yeast fermentation for bioethanol production. Three sequential types of by-products were analyzed-Ext1 (post hot-extraction), Ext2 (post fermentation), and Ext3 (post distillation)-and compared with Dat-Me. High Performance Liquid Chromatograp-Diode Array Detector-Mass Spectrometry (HPLC-DAD-MS) analysis allowed identifying 22 phenolic compounds, primarily cinnamic acid derivatives and glycosylated flavones such as luteolin and chrysoeriol. Fermentation increased total phenolic content from dry weight, while leading to an improved polysaccharide recovery (i.e., from 14.2% to 42.1% dry weight). Two polysaccharide fractions (F1 and F2) were isolated and characterized by H-1-NMR and Dynamic Light Scattering (DLS). F1 is a pectic polysaccharide, with a galacturonic acid content ranging from 24.2% (Ext3) to 52.2% (Dat-Me), a degree of methylation (DM) between 34.4 and 50.6%, and a degree of acetylation (DA) of 23.6-42.2%. F2 consists of a non-pectic polysaccharide, characterized by a low galacturonic acid content (5.6-6.8%) and a DM of 12.6-47.1%, but it is highly acetylated, with a DA ranging from 90.1 to 93.3%. DLS analysis confirmed fermentation-induced depolymerization, with molecular weights ranging from 6.6 & times; 10(4) to 8.5 & times; 10(5) KDa for both the fractions. Overall, Medjool date by-products obtained after bioethanol production represent a sustainable source of high-value phenolic antioxidants and polysaccharides with different structures suitable for future applications in food, pharmaceutical, and cosmetic formulations.

Several approaches have been proposed to support the panel test in virgin olive oil classification, but none of them is currently applied in olive oil companies. Aim of this study was the robust application of a chemometric model in a big olive oil company. The application on 244 samples of the PCA-LDA model developed in 2019, based on volatile profile by HS-SPME-GC-MS, gave unsatisfactory results, pointing out critical issues relating to the training-set, variable selection and validation. Therefore, a new t-test-FwS-LDA model was developed; it was based on a very wide dataset (approx. 1800 samples from 6 different production years) and on an algorithm for a stepwise selection of variables. The crucial role of the production year has been proven and included in the model. Ten volatile molecules were thus selected coming from both the lipoxygenase pathway and several virgin olive oil sensory defects. The new model was two-fold validated with 53 and 273 samples coming from production years belonging and not belonging to the training-set, respectively, with very satisfactory results (>90 % and 80 % correct classification, respectively). Finally, the study indicated that for routinary application of the model, year-by-year updating of training-set and variable selection is required.

Olive tree leaf is a phenol-rich, high-potential-value biomass that can be used to formulate food additives and supplements. Leaf phenolic content varies depending on numerous factors, like cultivar, geographical origin, year, and season of harvest. The aim of this research was to evaluate the variations in phenolic profile of four major Tuscan cultivars (Frantoio, Leccio del Corno, Leccino, and Moraiolo) over four different phenological phases and across two years. All 96 olive leaf samples were harvested from trees grown in the same orchard located in Florence. After drying, their phenolic profile was characterized using HPLC-DAD-MS, and the obtained data were processed by ANOVA, GA-LDA, and RF methods. A total of 25 phenolic derivatives were analyzed, with total contents ranging 16,674.0–50,594.3 mg/kg and oleuropein (4570.0–27,547.7 mg/kg) being the predominant compound regardless of cultivar, year, and season of harvest. Oleuropein and hydroxytyrosol glucoside showed inverse proportionality and similar behavior across years in all cultivars, and therefore were highlighted as main phenolic compounds correlated with the seasonal variability in studied cultivars. Interesting behavior was also pointed out for apigenin rutinoside. Application of GA-LDA and RF methods allowed pointing out the excellent performance of leaf phenols in discriminating samples based on cultivar, harvest year, and harvesting season.

Pomegranate peel is a by-product rich in ellagitannins, characterized by high molecular weight and low bioavailability. Working on several peel samples, we aimed to propose a single-step bicarbonate-assisted extraction to produce dry extracts with new phenol profiles rich in hydrolyzed tannins with potentially higher bioavailability. Compared to decoction, 0.6% sodium bicarbonate (NaHCO3) for 1 h extraction allowed 3.5-6.5-fold and 5.6-11.3-fold increase in ellagic acid and gallic acid, respectively, with total phenols ranging from 28.3% to 35.1% w/w. Simple pomegranate peel extraction with bicarbonate can be applied to prepare botanical extracts suitable for the food supplement market, thus increasing the value of this by-product.

This study provides a comparative analysis of the bioactive and volatile components in cold-pressed apricot (Prunus armeniaca L.) and peach (Prunus persica L.) kernel oils, emphasizing their potential applications in the food, nutraceutical and cosmetic industries. Both non-commercial cold-pressed oils and commercial samples were evaluated for their fatty acid profiles, volatile organic compounds, phenolic content, tocopherols, and other bioactive molecules. GC-FID analysis confirmed that both oils are rich in oleic and linoleic acids essential for their nutritional and skin-care properties, with varying levels of antioxidants, including tocopherols and poly-phenols, which contribute to their health-promoting properties. The analysis of volatile organic compounds, using HS-SPME-GC-MS, revealed distinct aroma profiles, with benzaldehyde as a key marker with a concentration in the range of 0.010-16.370 mg/kg, a wide range, that can be influenced by different factors including the botanical source as well as the processing and storage conditions. Furthermore, acrolein, a marker of lipid spoilage, was only detected in commercial oil samples, ranging from 0.005 to 0.385 mg/kg for apricot and 0.017-0.097 mg/kg for peach, respectively. Finally, thermogravimetric assessments showed non-significant differences between the samples, demonstrating significant stability of cold-pressed oils, while HPLC-ESI-MS identified 9 polar phenolic compounds. Cyanogenic glycoside analysis verified the safety of the oils, with undetectable or trace amounts in all samples. Thanks to these analytical methods, it was possible to identify the qualities of the oil and to assess whether non-commercial cold-pressed oils were more suitable for cosmetic or food use. Therefore, the results suggest that non-commercial apricot kernel oil, with its favorable PUFA/SFA ratio, is ideal for food applications, and both non-commercial oils can be used in cosmetic formulations.

Ulvan oligosaccharides, which exhibit greater solubility than high molecular weight ulvan, hold significant potential for the valorization of Ulva biomass. They have been shown to promote the growth of probiotic bacteria in fermentation models and have demonstrated protective and anti-inflammatory effects in animal models. The enzymatic hydrolysis of ulvan using the commercial ulvan lyase PLSV_3875 bypasses the challenge posed by the resistance of ulvanobiuronic acids to conventional acidic hydrolysis. This study aimed to develop a rapid, scalable, and efficient method for a complete ulvan hydrolysis under reduced salt concentrations, yielding oligosaccharides suitable for biological applications. The enzyme's robustness and kinetic parameters were evaluated. Enzyme activity was assessed at pH 6-10.5 with 0-100 mM Tris-HCl, carbonate or phosphate buffer, and 0-200 mM NaCl/KCl. Optimal conditions were identified as 3 mg/mL ulvan, 25 mM KCl, and 100 mM phosphate buffer at pH 7.5. The resulting oligosaccharides were characterized using 2D-NMR and UHPLC-MS/MS.

Consumers are increasingly willing to pay a premium for high-quality extra virgin olive oils (HQ-EVOOs) with specific sensory or nutraceutical properties, and originating from particular botanical or geographical origins. Regarding geographic origin, Italy is one of the main producers, with many local production areas, each characterized by its own distinctive typicity. The aim of this study is the chemical, sensory, and nutraceutical profiling of HQ-EVOO produced over two production years in Montespertoli (province of Florence) by 12 producers involved in the “MontEspertOlio” project, funded by the Tuscan Region. Oils were produced based on a production process previously defined and specifically applied to this territory. The shelf-life of the oil was also evaluated over a 12-month period. Legal quality parameters were analyzed according to EU regulation. Phenolic compounds, tocopherols, fatty acid composition, and volatile compounds were analyzed using HPLC-DAD, HPLC-FLD, HS-SPME-GC-MS, and GC-FID, respectively. Finally, sensory analysis was conducted using the Panel Test method. Results showed that Montespertoli HQ-EVOO is characterized by distinctive sensory and chemical traits that fully match consumer preferences, even across two production years characterized by different growing conditions. The shelf-life performance was excellent over 12 months, also showing a protective effect of greater bottle sizes against oxidation.

Background: Chemotherapy-induced neuropathic pain is a major side effect of antineoplastic treatment. This study investigated the neuroprotective potential of Acmella oleracea L. extracts containing the N-alkylamide spilanthol, phenolic acids, and glycosylated flavonoids. Methods: Hydroalcoholic extracts of aerial parts (AP) and roots (R) of in vitro seedlings were quali-quantitatively characterized by HPLC-DAD-MS and by 1H-NMR. Different concentrations (15–150 µg/mL) of AP and R were tested in SH-SY5Y cells differentiated into neurons exposed to oxaliplatin (10 µM), assessing cell viability (MTT), cytotoxicity (LDH), SOD activity, and expression of ATF-3, Ire1α, and Nf-H genes. To evaluate the impact on neuropathic pain, CD-1 mice were treated intraperitoneally with oxaliplatin (2.4 mg/kg), the effect of AP and R extracts (200–1200 mg/kg) were measured by the cold plate test. Results: AP extract was rich in phenols and alkylamides, whereas R extract showed higher phenolic levels but lower alkylamides content. Both extracts significantly reduced mortality and cytotoxicity and counteracted oxidative imbalance by enhancing SOD activity. Gene expression analysis confirmed their neuroprotective effects. In vivo, oxaliplatin induced a 50% reduction in pain threshold, while acute treatment with AP and R extracts dose-dependently alleviated neuropathic pain. Despite the lower spilanthol content in R extract, its efficacy was comparable to AP, suggesting an important role of phenolic compounds. Conclusions: Extracts from both aerial parts and roots of A. oleracea show promise in alleviating chemotherapy-induced neuropathy through mechanisms not solely related to spilanthol. Further studies to elucidate the contribution of phenolic components are desirable.

The polysaccharide ulvan has great potential for various industrial applications due to its unique composition and biochemical properties. The study aimed to optimize the extraction conditions (pH, extraction time, and Extractant/Solid (E/S) ratio) of ulvan employing Ulva lactuca samples collected in the Orbetello lagoon, Tuscany (Italy), using a Design of Experiments (DoE) approach. The extraction process was optimized through a screening phase followed by Response Surface Methodology (RSM) to maximize ulvan yield and selectivity of the extraction. The purified extracts were characterized by quantitative NMR for rhamnose (Rha) content, turbidimetric assay for sulfate content, and HPAEC-PAD for monosaccharide composition. The results indicated that a low pH value of 2.0, an extraction time of 105 min, and a high E/S ratio (60:1 mL/g) favored higher purity and yield of ulvan. Ulvan extracted under optimized conditions (9.27 % yield, 27.8 % Rha, and 20 % sulfate content) was further characterized by 2D NMR experiments and size exclusion chromatography. This optimized extraction method provides a basis for further exploration of ulvan's potential in food, pharmaceutical, and biotechnological applications.

Pomegranate peel is a byproduct of juice production rich in distinctive ellagitannins characterized by the gallagyl group. The resulting extract may be used as an oenological tannin, provided that it meets the minimum polyphenol concentration of 65% w/w set by the International Organization of Vine and Wine (OIV) and contains low levels of pectin. The aim was to improve conventional heat-assisted extraction (HAE) to obtain extracts enriched in polyphenols and lowered in pectic polysaccharides. Therefore, HAE, Low-Temperature Hydroalcoholic Extraction (LTHE) and Solid-State Fermentation with Saccharomyces cerevisiae sp. combined with LTHE (SSF + LTHE) were compared. LTHE yielded the lowest polysaccharide content (0.2-1% w/w), whereas SSF + LTHE allowed for an average 40% increase of polyphenols and a 70% reduction in polysaccharides compared to HAE. Only SSF + LTHE allowed the recovery of total polyphenol percentages above the OIV threshold, highlighting the promising potential of solid-state fermentation-assisted extraction to produce polyphenol-rich extracts from pomegranate peel.

An innovative, non-destructive, and rapid method was introduced to quantify hydroxytyrosol content in olive leaves by means of advanced photonic technologies and chemometrics. At the heart of this approach is a custommade, pocket-sized fluorometer with LED excitation at 375 nm and a miniaturized spectrometer for emission detection in the visible range, designed to measure intact olive leaves with ease and precision. Four Italian olive cultivars, namely Frantoio, Leccino, Leccio del Corno, and Moraiolo, were studied, with nine leaf samplings conducted over the 2022-2024 period. Simultaneously, hydroxytyrosol concentration was measured using HPLC-DAD-MS on the same samples to establish the reference data. Chemometric analysis was applied to the spectroscopic and HPLC-DAD-MS results. The Orthogonal Partial Least Squares (OPLS) regression method was successfully used to build a predictive model to quantify hydroxytyrosol concentrations ranging from 200 to 7000 mg/kg. The model achieved very good regression coefficients of 0.9 for calibration and 0.83 for cross-validation, with root mean square errors of 600 mg/kg and 720 mg/kg, respectively. This non-destructive method, combined with the portability of the pocket-sized fluorometer, is an innovation for high-throughput screening of olive leaves. It offers significant potential for evaluating olive leaves before their use in tea production or as raw material for dietary supplements, making it highly appealing for both the agriculture and health industries.

Cocoa bean shells (CBS), a major chocolate industry by-product, rich in dietary fibres and bioactive compound, represent a promising target for upcycling in sustainable food systems. This study evaluates the CBS lipidic fraction extracted by Soxhlet using four apolar solvents: dichloromethane (DE), petroleum ether (PE), diethyl ether (EE), and hexane (HE). The extracts were analysed for yield and chemical composition, including phenolics, methylxanthines, phytosterols, tocopherols, and volatile organic compounds (VOCs). Results showed a significant variation in extraction yields, with DE providing the highest yield (8.05 g/100 g of CBS), compared to others (5.13-5.92 g/100 g of CBS). The phenolic content (3.57-38.2 mg/kg of lipid extract) varied depending on the solvent used, with protocatechuic acid being the most abundant compound (up to 32.2 mg/kg of lipid extract obtained with EE). Brassicasterol was detected in all the extracts (337-520 mg/kg of lipid extract), confirming the presence of valuable phytosterols. Additionally, 115 VOCs were quantified across all samples, with great variation between solvents (23.4-116.5 mg/kg of lipid extract). This study highlights CBS as a valuable source of bioactive lipids and supports their valorisation for the recovery of high-value compounds. These results pave the way for the development of natural ingredients applicable in functional foods, nutraceuticals, and cosmetics, contributing to agro-industrial waste reduction and promoting circular economy strategies.