Saccharomyces cerevisiae is commonly used for the production of alcoholic beverages, including cider. In this study, we examined indigenous S. cerevisiae and S. uvarum strains, both species commonly found in cider from Hardanger (Norway), for their strain-specific abilities to produce volatile and non-volatile compounds. Small-scale fermentation of apple juice with 20 Saccharomyces strains was performed to evaluate their aroma-producing potential as a function of amino acids (AAs) and other physicochemical parameters under the same experimental conditions. After fermentation, sugars, organic acids, AAs, and biogenic amines (BAs) were quantified using the HPLC–UV/RI system. A new analytical method was developed for the simultaneous determination of nineteen AAs and four BAs in a single run using HPLC–UV with prior sample derivatization. Volatile compounds were determined using HS-SPME-GC-MS. Based on 54 parameters and after the removal of outliers, the nineteen strains were classified into four groups. In addition, we used PLS regression to establish a relationship between aroma compounds and predictor variables (AAs, BAs, organic acids, sugars, hydrogen sulfide (H₂S) production, CO₂ release) of all 19 strains tested. The results of the VIP show that the main predictor variables affecting the aroma compounds produced by the selected yeasts are 16, belonging mainly to AAs.

Our investigation delves into the previously uncharted territory of cider composition from Norway. This study aimed to obtain an overview of the qualitative and quantitative compositions of general chemical parameters, polyphenols (individual and total expressed as gallic acids equivalents), selected esters, and selected C6-alcohols in ciders with the PDO label Cider from Hardanger. In total, 45 juice and cider samples from the fermentation process were collected from 10 cider producers in Hardanger in 2019, 2020, and 2021. Individual sugars, acids, ethanol, and 13 individual phenols were quantified using HPLC-UV/RI. Seven ethyl esters of fatty acids, four ethyl esters of branched fatty acids, ten acetate esters, two ethyl esters of hydroxycinnamic acids, and four C6-alcohols were quantified using HS-SPME-GC-MS. For samples of single cultivars (‘Aroma’, ‘Discovery’, ‘Gravenstein’, and ‘Summerred’), the sum of the measured individual polyphenols in the samples ranges, on average, from 79 to 289 mg L⁻¹ (the lowest for ‘Summerred’ and highest for ‘Discovery’ and ‘Gravenstein’). Chlorogenic acid was the most abundant polyphenol in all samples. Ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, ethyl isobutyrate, ethyl 2-methylbutyrate, isoamyl acetate, and hexanol were present at concentrations above the odour threshold and contributed to the fruity flavour of the Cider from Hardanger.

This study investigated the chemical aromatic profiles of 60 volatile organic compounds (VOCs), including 21 terpenes, 6 norisoprenoids, 6 volatile phenols, 4 C-6 alcohols, and 23 esters, in Graševina wines originating from Slavonia and Podunavlje sub-regions (Croatia). Headspace solid-phase microextraction gas chromatography coupled with mass spectrometric detection (HS-SPME-GCMS) was used to assess 60 VOCs, and a novel HS-SPME-GCMS method for the determination of terpenes and norisoprenoids was developed and validated. Statistical analysis found no significant differences between VOCs present in Graševina wines and others wines (Chardonnay and Pinot gris); nonetheless, comparison of VOC fingerprints between different wines from the same producer showed that four terpenes (α-terpinene, γ-terpinene, 1,4-cineol, and 4-terpineol) were present in higher amounts in Graševina wines when compared to other wines. By exclusively examining VOCs within Graševina wines through the utilization of a heatmap and hierarchical clustering, a distinct visualization of samples and VOCs emerged, highlighting the vintage effect. Simultaneously, a partial least squares discriminant analysis model was developed with a confidence interval of 95%, demonstrating a noticeable distinction between samples originating from the western and eastern regions. Furthermore, by employing the heatmap using only VOCs from the terpene and norisoprenoidic groups, a clear separation of samples into two groups was observed based on geographical origin; namely, higher concentrations of norisoprenoids and terpenes were observed in the Slavonia sub-region. These results suggest that terpenes and norisoprenoids to a lower extent may be valuable aromatic markers for the regional typicality of Graševina wines.

Botrytis cinerea Pers., the causal agent of grey mould, causes major economic losses in viticulture by reducing grape and wine quality and yield. Antagonistic yeasts that release bioactive volatile organic compounds (VOCs) represent a sustainable alternative to synthetic fungicides. Here, VOCs produced by Pichia guilliermondii strain ZIM624 were identified and assessed for antifungal activity against B. cinerea. 65 VOCs—including higher alcohols, volatile phenols, esters, and terpenes—were detected using two newly developed and validated analytical methods combining automated headspace solid-phase microextraction with gas chromatography–mass spectrometry. A total of 13 VOCs were selected for the bioassays. Fumigation assays demonstrated that terpenes (citronellol, geraniol, nerol, α-terpineol, and linalool) were the most effective inhibitors of B. cinerea mycelial growth (EC₅₀ = 6.3–33.9 μL/L). Strong inhibition was also observed for 4-vinylphenol and isoamyl acetate. In vivo assays confirmed that exposing infected grape berries to P. guilliermondii VOCs significantly reduced grey mould incidence. These results highlight the potential of P. guilliermondii ZIM624 volatiles as natural biofumigants for the eco-friendly management of B. cinerea in grapes. Future research should focus on optimising VOC production, evaluating efficacy under field conditions, and developing formulations for practical application in vineyards and post-harvest storage. Additionally, investigating potential synergistic effects of VOC combinations could lead to more effective biocontrol strategies.

Background/Objectives: Adeno-associated viruses (AAVs) are widely used gene therapy vectors; yet their physicochemical stability and chromatographic behavior are highly sensitive to the solution conditions they are in. Effective separation of full (F), empty (E), and partially filled (P) capsids—most commonly achieved by anion exchange (AEX) chromatography—is essential for standard analytical characterization, process development, and product safety. However, conventional AEX methods rely on low-conductivity alkaline mobile phases with low salt, which promote capsid binding and therefore higher resolution, at the expense of structural stability. Conversely, formulations such as near-neutral buffers might preserve capsid integrity but often impair AEX retention and separation resolution. Methods: Here, we extend a mechanistic investigation using AAV8 capsids as a model system, focusing on detailed capsid interactions with strong AEX, and present novel AAV8 separation strategies on a weak AEX stationary phase. Results: By systematically varying buffer pH and ionic strength, we identify operational regimes that balance capsid stability with chromatographic separation efficiency. In parallel, we introduce an integrated two-dimensional (2D) in-line buffer exchange configuration that decouples AEX performance from sample formulation, enabling robust separation of stability-optimized, high-salt matrices without off-line desalting. Conclusions: By elucidating the roles of capsid charge modulation, ligand physicochemical properties, and local microenvironmental buffering, this study establishes practical design principles for stability-preserving chromatography. It lays a foundation for more reliable analytical and future preparative AAV workflows.