Headspace Solid-phase Microextraction Research Articles

Study on volatile metabolites and microbial community in the fermentation process of black tea (Jiuqu hongmei tea)

Jiuqu hongmei tea (JQHM tea), unique to Zhejiang Province, China, is celebrated for its vibrant, delightful taste and distinctive flavor profile. This research explored the relationship between the tea's characteristic flavor during fermentation and the dynamics of microbial communities. Volatile organic compounds (VOCs) were analyzed using Headspace Solid-Phase Microextraction coupled with Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS) at three key stages: machine kneading (MK, 0.5 hours), primary fermentation (F1, 1.5 hours), and secondary fermentation (F2, 3 hours). Concurrently, microbial communities were characterized through high-throughput sequencing. The study identified 12 significant VOCs and pinpointed 10 core bacterial and 9 core fungal genera. These findings revealed the intricate changes in aroma compounds and microbial populations throughout JQHM tea's fermentation process, highlighting the interplay between key aroma compounds and principal microbial genera. This knowledge provided a crucial theoretical foundation for refining the tea's processing stages and enhancing its flavor quality.

Comparative volatiles profiling of two marjoram products via GC-MS analysis in relation to the antioxidant and antibacterial effects.

Marjoram (Origanum majorana L.), also known as "sweet marjoram" or "sweet oregano" is a Mediterranean herbaceous perennial herb cultivated in Egypt and widely consumed as an herbal supplement for treatment of several ailments. The main goal of this study was to assess volatiles' variation in marjoram samples collected from two different widely consumed commercial products using two different extraction techniques viz. head space solid phase microextraction (HS-SPME) and petroleum ether using gas chromatography mass spectrometry (GC-MS) analysis and multivariate data analysis. A total of 20 major aroma compounds were identified in samples extracted with HS-SPME found enriched in monoterpene hydrocarbons and oxygenated compounds. The major volatiles included β-phellandrene (20.1 and 14.2%), γ-terpinene (13.4 and 11.7%), 2-bornene (12.3 and 11.5%), p-cymene (9.8 and 4.6%) terpenen-4-ol (16.4 and 7.5%), sabinene hydrate (16.02 and 8.8%) and terpineol (4.2 and 3.2%) in MR and MI, respectively. Compared with HS-SPME, 51 aroma compounds were identified in marjoram samples extracted with petroleum ether, found more enriched in aliphatic hydrocarbons (42.8 and 73.8%) in MR and MI, respectively. While a higher identification score was observed in the case of solvent extraction, SPME appeared to be more selective in the recovery of oxygenated terpenes to account more for marjoram aroma. Multivariate data analysis using principal component analysis (PCA) revealed distinct discrimination between volatile composition of both marjoram samples. The total phenolic and flavonoid contents in marjoram samples were at (111.9, 109.1µg GA/mg) and (18.3, 19.5µg rutin eq/mg) in MR and MI, respectively. Stronger antioxidant effects were observed in MR and MI samples with IC50 at 45.5 and 56.8µg/mL respectively compared to IC50 6.57µg/mL for Trolox as assayed using DPPH assay. Moderate anti-bacterial effect was observed in MR and MI samples and expressed as a zone of inhibition mostly against Bacillus subtilis (16.03 and 15.9mm), B. cereus (12.9 and 13.7mm), Enterococcus faecalis (14.03 and 13.97mm), and Enterobacter cloacae (11.6 and 11.6mm) respectively.

Stimulation of Saliva Affects the Release of Aroma in Wine: A Study of Microbiota, Biochemistry, and Participant Origin.

Saliva influences the release of aroma in the oral cavity. The composition of human saliva varies depending on stimulation and host's origin; however, the compositional differences of saliva and their influences on aroma release have not been fully evaluated. In this study, we recruited 30 healthy adults (15 Australians and 15 Chinese) and collected saliva samples at three stages: before, during, and after stimulation. Salivary samples were characterized by the flow rate, total protein concentration, esterase activity, microbiome composition by full-length 16S rRNA gene sequencing, and the ability to release aroma from wine by headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS). Differences in salivary composition and specific wine volatiles were found between Australian and Chinese participants and among the three stimulation stages. Significant correlations between the relative abundance of 3 bacterial species and 10 wine volatiles were observed. Our results confirm the influence of participant's geographic origin and stimulation on the salivary composition, highlighting the role of salivary components, especially salivary bacteria, on the release of aroma from wine.

Characterization of the aroma and flavor profiles of guava fruit (Psidium guajava) during developing by HS-SPME-GC/MS and RNA sequencing

The flavor of guava, an important tropical fruit, is influenced by secondary metabolites. However, the mechanisms and processes underlying flavor formation in guava remain unclear. In this study, dynamic changes in volatile organic compounds (VOCs), sugars, and organic acids in guava peel and flesh across different developmental stages were investigated using headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC–MS) and high-performance liquid chromatography (HPLC). Here, we identified 90 VOCs, three sugars and eight organic acids. The dynamics of VOCs differ between the flesh and peel. The early developmental stages are more critical in influencing the variation of VOCs in the flesh, while VOC changes in peel occur more progressively across the developmental stages. By integrating transcriptomic and metabolomic analyses, we identified several key genes involved in VOC, sugar, and acid metabolism. This is the first study to describe the expression patterns of these genes throughout guava development, providing new insights into guava flavor development.

Influence of Terroir on the Grain Composition, and Volatile Profile of Irish Grain (Wheat) New Make Spirit

Terroir refers to the combination of environmental factors, such as climate, soil, and agricultural practices, that shape the characteristics of a crop, contributing to the unique qualities of the final product. The concept has been traditionally linked to wine, but some recent findings suggest that it also holds importance for distilled spirits. The expanding Irish distilling sector is shifting towards local raw materials such as wheat and rye, driven by regulatory changes, economic benefits, and consumer demand for sustainable local products. This research examines the effects of wheat variety, geographical location, and harvest year on grain composition and volatile composition of the new make spirit. For this study, twenty lab-scale wheat whiskey samples were produced from five different wheat varieties grown at two different locations in Ireland over two consecutive years. The wheat samples were analysed for grain composition and the volatile profiling of new make spirit samples by headspace solid-phase microextraction (HS-SPME) followed by gas chromatography–mass spectrometry (GC-MS). A total of fifty-one volatile compounds were detected, with ethanol, ethyl acetate, phenyl ethyl alcohol, and 3-methyl-1-butanol being predominant. Principal component analysis revealed that both the harvest year and geographical location moderately influenced the volatile compound distribution of the new make spirit, which is explained by a 43.25% variance. ANOVA analysis revealed that grain composition was significantly influenced by harvest year, location, and wheat variety. The 2020 samples showed higher protein and β-glucan content, whereas samples from the location Tipperary had higher starch content. This study indicates that terroir—specifically seasons (year) and geography (location)—affects the characteristics of wheat-based Irish whiskey, highlighting opportunities for distillers to differentiate their products by leveraging local environmental factors.

Comparative Investigation of Aroma-Active Volatiles in ("Ruixue", "Liangzhi", "Crystal Fuji," and "Guifei") Apples by Application of Gas Chromatography-Mass Spectrometry-Olfactometry (GC-MS-O) and Two-Dimensional Gas Chromatography-Quadrupole Mass Spectrometry (GC × GC-qMS) Coupled with Sensory Molecular Science.

Aroma dramatically impacts the overall flavor profiles and consumer acceptance; therefore, it is necessary to conduct a comprehensive analysis of the aroma characteristics of apples. In this study, the aroma differences among four popular apple varieties ("Ruixue", "Liangzhi", "Crystal Fuji," and "Guifei") were compared using two extraction methods (headspace-solid phase microextraction, and solvent-assisted flavor evaporation) coupled with gas chromatography-mass spectrometry-olfactometry (GC-MS-O) and two-dimensional gas chromatography-quadrupole mass spectrometry (GC × GC-qMS). A total of 82 odorants were identified via GC-MS-O, and 143 volatiles were identified by GC× GC-qMS. Among them, 41 key aroma-active compounds (butanal, ethyl acetate, 3-methylbutanal, methyl butanoate, 2-methylpropyl acetate, ethyl 2-methylbutanoate, ethyl 3-methylbutanoate, butyl acetate, hexanal, 2-methylbutyl acetate, 1-butanol, 2-methylpropyl butanoate, 3-methylbutyl acetate, (E)-2-hexenal, butyl butanoate, butyl 2-methylbutanoate, hexyl acetate, hexyl butanoate, hexyl, 2-methylbutanoate, 1-octen-3-ol, 3-methylthiopropanol, 1,3-octanediol, linalyl acetate, and so on) with high odor activity values (OAVs) and AI value (odor activity values ≥1 or aroma intensity ≥3) were identified. Partial least-squares-discriminant analysis showed that Ruixue exhibited a high "fruity" note, Guifei and Crystal Fuji had the greatest "wood," "floral," and "sweet" notes, while Liangzhi presented a significant "green" note. This study provided flavor chemistry support for the apple quality control and production.

Study on Evaluation of Fruit Aroma of Plum Variety Resources Based on Headspace Solid-Phase Microextraction Combined with Gas Chromatography-Mass Spectrometry.

To explore the characteristics of and variations in aroma components across different plum varieties and maturity stages, this study employed headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). This method was used to systematically analyze the aroma components of 12 early-maturing, 15 medium-maturing, and 11 late-maturing plum varieties. The variations in volatile components among these three germplasm types were then compared using multivariate statistical methods. The examination revealed that 138 aromatic components were meticulously identified and quantified, such as 26 aldehydes, 63 esters, 13 ketones, 30 alcohols, and six other compounds. Thirteen main aroma compounds including acetic acid hexyl ester, (Z)-3-hexen-1-ol acetate, hexanal, 1-hexanol, 3-hexenal, butanoic acid butyl ester, (E)-2-hexen-1-ol, hexanoic acid butyl ester, propanoic acid butyl ester, (E)-2-hexenal, L-.alpha.-terpineol, (Z)-2-hexen-1-ol acetate, and 1-butanol were considered dominant. The orthogonal partial least squares discriminant analysis (OPLS-DA) combined with variable importance projection (VIP) results showed that 24 differential aroma compounds were screened out from 38 varieties of plum fruits based on their differences in aroma components, which can be used to distinguish plum fruits at different ripening times. Twenty-four aroma-contributing compounds were identified based on their odor activity values (OAVs). Among these, 14 key aroma components with OAVs ≥ 10 were highlighted. In summary, the aroma compounds of early- and late-maturing plum germplasm exhibited rich diversity, with significant differences in aroma components between plums of varying maturity and germplasm. These differences can serve as indicators for identifying different plum germplasm.

Comprehensive two dimensional gas chromatography – Time of flight mass spectrometry (GC×GC-TOFMS) for the investigation of botanical origin of raw spirits

Comprehensive two dimensional gas chromatography – time of flight mass spectrometry (GC × GC-TOFMS) with sample introduction using headspace solid phase microextraction (HS-SPME) was used for the botanical classification of raw spirits obtained from C3 (corn and sorghum) and C4 (rye, wheat and potato) plants. 45 spirit samples representing these raw materials (10 spirits produced from rye, corn, wheat and potato, and 5 from sorghum) were analyzed. Volatile compounds profiles were compared by PCA, and after removal of outliers samples were subjected to the classification model. OPLS-DA model was built (R2Y = 0.924 Q2Y = 0.895) that enabled clear separation of all tested spirits of different botanical origin. The model was validated with training and testing sets and 100 % correct assignment was achieved. GC × GC proved to be a method that not only can be used as a tool for botanic origin of raw spirits, but also provides detailed information of volatile fermentation by-products, characteristic for particular spirit.

Improving Quality Profiles of Summer-Autumn Tea Through the Synergistic Fermentation with Three Fungal Strains

The co-fermentation of summer-autumn tea with Aspergillus niger and Eurotium cristatum effectively reduces bitterness and astringency, enhances flavor and quality, and increases market appeal and economic value. This study investigated the dynamic changes in metabolites, flavor characteristics and quality indicators of summer-autumn tea during fermentation. Using headspace solid-phase microextraction gas chromatography (HS-SPME-GC-MS), 96 key metabolites were identified, significantly enhancing the tea's aroma and flavor. On the fourth day of fermentation, the content of ester catechins (EGCG, ECG) decreased by 79.44% and 79.57%, respectively, while non-ester catechins (EGC, C, EC) increased by 175.47%, 228.18%, and 138.73%, respectively. Additionally, the gallic acid content increased by 755.58%, along with rises in free amino acids and soluble sugars. Sensory evaluation focused on taste, soup color, and aroma, resulting in a total score of 80.67. The intensity of astringency, bitterness, and sourness in the tea infusion gradually decreased, while sweetness, umami, and aroma intensified. These findings underscore the significant role of the synergistic action of the three strains in developing the unique flavor profile of summer-autumn tea during fermentation. This research lays the foundation for quality control and resource development in the processing of summer-autumn tea.

Effects of compound fermentation of lactic acid bacteria IMAUJBP3 and IMAUJBR3 on the characteristic flavors and metabolites of mutton fermented sausages

Lactobacillus plantarum IMAUJBP3 and Limosilactobacillus reuteri IMAUJBR3 were used to prepare starter cultures. Through the electronic nose (E-nose), it was found that the compound fermentation can enhance the aromatic components in the product. Further analyses using Headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPMEGC–MS) and metabolomics revealed that the complex fermentation affected the oxidative breakdown of amino acids and lipids. HS-SPMEGC-MS identified 72 flavour compounds and found that the Mix group had the highest amount of ester compounds relative to the ZR group. In addition, of the 1402 metabolites identified by metabolomics, dihydroxyacetone phosphate was upregulated and most abundance in the Mix group. Dihydroxyacetone phosphate promotes the metabolism of carbohydrates and lipids, and affects the oxidative decomposition of amino acids and lipids, resulting in differences in flavor among meat products inoculated with different starter cultures. At the same time, it was also found that compound fermentation could down-regulate the compounds affecting product quality such as histamine, putrescine and phenethylamine. Overall, this study found that JBP3 and JBR3 in combination could improve the flavour quality of fermented lamb sausages. These findings may provide a good strain resource for improving the quality of lamb sausage.

Phytochemical Characterization Utilizing HS-SPME/GC-MS: Exploration of the Antioxidant and Enzyme Inhibition Properties of Essential Oil from Saudi Artemisia absinthium L.

Background/Objectives: This study aimed to analyze the chemical composition and biological activities of Artemisia absinthium L. essential oil, focusing on its antioxidant and enzyme inhibition (α-amylase and urease) properties. Additionally, in vitro pharmacokinetic and pharmacodynamic evaluations were conducted through in silico molecular docking and BOILED-Egg models to assess its therapeutic potential and its potency in treating oxidative-stress-related diseases. Methods: The essential oil was isolated by the hydrodistillation (HD) of fresh plant material, and volatiles released from dried plant material were sampled via headspace solid-phase microextraction (HS-SPME), followed by a phytochemical profiling analysis through the GC-MS tool. Antioxidant capacity was assessed using DPPH, ABTS, FRAP, and nitric oxide scavenging assays, while enzyme inhibition activities were tested against α-amylase and urease. Molecular docking and BOILED-Egg models were used to evaluate compound interactions with NADPH oxidase and predict pharmacokinetic behavior, respectively. Results: HS-SPME and HD yielded 46 and 25 compounds, respectively, primarily terpenoids represented by camphor (26.4%) and cis-davanone (18.0%) in HS-SPME, while in the HD essential oil, cis-davanone (60.2%) and chamazulene (10.8%) were most prevalent. The antioxidant assays showed a strong activity, with a total antioxidant capacity of 821.8 mg ascorbic acid Eq/gm. The essential oil inhibited urease by 86.7% and α-amylase by 81.8%. Molecular docking showed strong binding affinities with NADPH oxidase, supporting the antioxidant results. Conclusions:A. absinthium EO demonstrated potent antioxidant and enzyme inhibitory activities, suggesting its therapeutic potential for treating enzyme-related disorders like diabetes mellitus and its possible use as a cure for many oxidative-stress-related diseases, thus validating the folkloric use of this plant.

Defective porous urchin-like ZnO/NiO microspheres-coated solid-phase microextraction fiber for analysis of trace polychlorinated biphenyls in milk.

Owing to the high lipophilicity of polychlorinated biphenyls (PCB), they easily accumulate in dairy products. Although usually present at very low levels, they pose a serious threat to human health. Therefore, developing a sensitive and reliable method for detecting PCB in dairy products is crucial. Herein, Herein, a metal-organic framework (MOF) material named with bimetallic nodes and double ligands was prepared as a precursor using a one-pot hydrothermal method. Defective porous urchin-like ZnO/NiO, derived from these MOF-based precursors (ZnNi-MOF-NH2) as a sacrificial template, was synthesized via pyrolysis to remove heat-sensitive ligands. To the best of our knowledge, this urchin-like nanostructured ZnO/NiO hybrid was utilized as a solid-phase microextraction (SPME) coating for the first time. Headspace SPME (HS-SPME) was developed for non-contact extraction of PCB in milk prior to gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis. Under optimal conditions, the HS-SPME-GC-MS/MS method exhibited a wide linear range (0.01-1000ng·L-1), low limits of detection (0.003-0.025ng·L-1), and high enrichment factors (5714-9906). Additionally, the performance of the ZnO/NiO SPME fiber coating showed no noticeable decrease after 175 uses. The method was applied to trace PCB analysis in milk samples, yielding recoveries of 70.3-114.1%. The ZnO/NiO derived from MOF-based material provides a promising candidate for SPME coatings to extract PCB and other analogs.

Combined Analysis of Grade Differences in Lapsang Souchong Black Tea Using Sensory Evaluation, Electronic Nose, and HS-SPME-GC-MS, Based on Chinese National Standards.

Tea standard samples are the benchmark for tea product quality control. Understanding the inherent differences in Chinese national standards for Lapsang Souchong black tea of different grades is crucial for the scientific development of tea standardization work. In this study, Lapsang Souchong black tea of different grades that meet Chinese national standards was selected as the research object. The aroma characteristics were comprehensively analyzed through sensory evaluation, electronic nose, and HS-SPME-GC-MS (headspace solid-phase microextraction gas chromatography-mass spectrometry). The findings indicate that the higher-grade Lapsang Souchong has a higher evaluation score. The results of electronic nose analysis indicate that the volatiles with differences in tea of different grades were mainly terpenoids and nitrogen oxides. The results of HS-SPME-GC-MS analysis show that the odor characteristics of the super-grade samples are mainly floral and fruity, and these substances mainly include D-Limonene, 3,7-dimethyl-1,6-octadien-3-ol and 3-Hydroxymandelic acid, and ethyl ester. The primary aroma characteristics of the first-grade samples are floral, sweet, woody, and green, with key contributing compounds including 2-Furanmethanol, 1-Octen-3-ol, and 5-ethenyltetrahydro-α,α,5-trimethyl-cis-, 4,5-di-epi-aristolochene. The main aroma characteristics of the second-grade samples are green, herbal scent, and fruity, and the main substances include 3,7-dimethyl-1,6-octadien-3-ol, 2,3-dimethylthiophene, Dihydroactinidiolide, and Naphthalene-1-methyl-7-(1-methylethyl)-. It is worth noting that the second-grade samples contain a large amount of phenolic substances, which are related to the smoking process during processing. This study lays a solid foundation for the preparation of tea standard samples and the construction of the tea standard system.

Impact of Mild Field Drought on the Aroma Profile and Metabolic Pathways of Fresh Tea (Camellia sinensis) Leaves Using HS-GC-IMS and HS-SPME-GC-MS.

Aroma plays a pivotal role in defining tea quality and distinctiveness, and tea producers have often observed that specific drought conditions are closely associated with the formation and accumulation of characteristic aroma compounds in tea leaves. However, there is still limited understanding of the differential strategies employed by various tea cultivars in response to drought stress for the accumulation of key volatile aroma compounds in fresh tea leaves, as well as the associated metabolic pathways involved in aroma formation. In this study, two widely cultivated tea cultivars in China, Fuding Dabai (FD) and Wuniuzao (WNZ), were examined to assess the impact of mild field drought stress on the composition and accumulation of key volatile aroma compounds in fresh leaves using headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS) and headspace solid phase micro-extraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) technologies. Results revealed that drought stress led to a substantial increase in the diversity of volatile compounds (VOCs) in FD, while WNZ exhibited a notable rise in low-threshold VOC concentrations, amplifying sweet, floral, fruity, and earthy aroma profiles in post-drought fresh leaves. Through partial least squares discriminant analysis (PLS-DA) of HS-GC-IMS and HS-SPME-GC-MS data, integrating variable importance projection (VIP) scores and odor activity values (OAVs) above 1, 9, and 13, key odor-active compounds were identified as potential markers distinguishing the drought responses in the two cultivars. These compounds serve as crucial indicators of the aromatic profile shifts induced by drought, providing insights into the differential metabolic strategies of the cultivars. Additionally, KEGG enrichment analysis revealed 12 metabolic pathways, such as terpenoid biosynthesis, fatty acid synthesis, cutin, suberine, and wax biosynthesis, and phenylalanine metabolism, which may play crucial roles in the formation and accumulation of VOCs in tea leaves under drought stress. These findings provide a comprehensive framework for understanding the cultivar-specific mechanisms of aroma formation and accumulation in tea leaves under mild drought conditions.

Determination of Residual Dimethyl Sulfate in Cephalosporin Using HS-SPME/GC-MS.

Dimethyl sulfate (DMS) is widely used in manufacturing process but because of its genotoxicity nature, it should be monitored at trace levels (1μg/mL). It is complicated and difficult to quantify DMS in cephalosporin with traditional method. Herein, a method for quantifying residual DMS in cephalosporin was developed, without complex sample preprocessing, no need for a large amount of solvent, employing headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). Compared with polydimethylsiloxane (PDMS)/divinylbenzene and polyacrylate fibers, PDMS was more suitable for absorbing DMS. The research showed that the PDMS fiber should be changed after 50 adsorption-desorption cycles. Linear regression analysis of the calibration curve demonstrated a robust linear relationship, with R2 of 0.999, across a concentration range of 0.25 to 4.0μg/mL. The method underwent rigorous validation for specificity, linearity, precision and accuracy. This method was proven effective in measuring DMS in complex matrices. The limits of detection and quantification for this method is 0.05 and 0.25μg/mL, respectively, which has room for improvement.

Identification of potential volatile markers for characterizing Argentine wine vinegars based on their production process

In Argentina, the production of quality wine vinegars has been barely exploited. Currently, most of the marketed vinegars are produced using rapid industrial fermentation systems obtaining vinegars with a high production rate, but with low quality and few organoleptic nuances. This work aims to study the volatile profile by headspace solid phase microextraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC–MS) and chemometrics to differentiate Argentinian wine vinegars according to their production process (industrial or traditional). A total of 92 volatile compounds were identified on the samples by using a strategy based on volatile profile processing using PARADISe® software. The complete volatile profile of all the samples was submitted to partial least squares discriminant analysis (PLS-DA) for the selection of variables with importance in the projection (VIPs). Thus, 37 volatile compounds with the potential to be markers of the manufacturing process were selected. The results obtained revealed, for the first time, the volatile profile of Argentine wine vinegars showing that the compound groups that, on average, exhibited higher relative areas were esters, acids, and alcohols. Thus, certain acids, aldehydes and terpenes were noticeably present in industrial wine vinegar. Conversely, the alcohols were strongly associated with traditional ones. In the case of esters, they were connected to the different types of wine vinegars. These compounds could be considered as potential volatile markers of quality and authenticity of these types of vinegars.

Composition of Higher Alcohols in Different Alcoholic Beverages and Their Metabolic Dynamics in Bama Pigs.

The unique flavour contribution of higher alcohols in alcoholic beverages has received growing attention; however, there is a dearth of information on their in vivo metabolic kinetics. In this study, the composition and content of higher alcohols in different alcoholic beverages from Chinese Baijiu and Lujiu were studied via in vivo analysis using Bama pigs to elucidate the mechanisms for intoxication of alcohol in vitro and in drinkers. Direct injection combined with gas chromatography-mass spectrometry (GC-MS) were used to accurately quantify a total of 14 higher alcohols in five alcoholic beverages. Based on the external standard method, a total content of 289.37-938.33 mg/L was detected, mainly 1-butanol, 3-methyl-1-butanol, 1-hexanol, 2-methyl-1-propanol and 2-butanol. Then, headspace solid-phase microextraction (HS-SPME) and solid-phase extraction (SPE) combined with GC-MS analysis strategy, respectively, were adopted to continuously monitor the changes in the concentrations of ethanol and 11 higher alcohols in the blood within 24 h after gavage of different alcoholic beverages, and the key pharmacokinetic parameters were analysed. The peak concentration (Cmax) and area under curve (AUC) of blood higher alcohols were significantly lower than those of ethanol (p < 0.05), accompanied by a later peak time (Tmax) and a larger apparent clearance rate (CL_F), and there were certain differences between the same higher alcohols in different alcoholic beverages and between different higher alcohols in the same alcoholic beverage. This work provides valuable insights into the metabolism of alcoholic beverages.

The effect of indigenous non-Saccharomyces yeasts on the volatile profile of Maraština wine: Monoculture versus sequential fermentation

Non-Saccharomyces yeasts are presented as a new promising tool in winemaking to enhance aroma complexity in fermentation with Saccharomyces yeasts. Indigenous yeasts are recognized for their better adaptation to environmental conditions and for highlighting the unique terroir impact on wine aroma characteristics. To study the individual impact of ten indigenous non-Saccharomyces yeasts on the primary metabolites and volatile compounds of wine from autochthonous Croatian Maraština variety, Hyphopichia pseudoburtonii, Metschnikowia chrysoperlae, Metschnikowia sinensis/shanxiensis, Metschnikowia pulcherrima, Lachancea thermotolerans, Hanseniaspora uvarum, Hanseniaspora guilliermondii, Hanseniaspora pseudoguilliermondii, Pichia kluyveri, and Starmerella apicola were inoculated in sterile grape juice in monoculture fermentations. Additionally, seven of them were also studied in sequential fermentation of sterile grape juice with commercial Saccharomyces cerevisiae. A targeted approach was used for the identification of volatile compounds via headspace solid-phase microextraction coupled with gas chromatography and a mass spectrometer. P. kluyveri and H. uvarum produced higher total concentrations of terpenes in sequential versus monoculture fermentation. Sequential fermentation of M. chrysoperlae, L. thermotolerans, and P. kluyveri with S. cerevisiae resulted in higher production of C13-norisoprenoids. The esters concentration was higher in monocultures for L. thermotolerans, H. uvarum, and H. guilliermondii, whereas other isolates showed higher concentrations in sequential fermentations. The results highlighted different indigenous yeast metabolisms and provided promising insights into potential new non-Saccharomyces starter cultures as a first step in their selection, with several species characterized in terms of their potential effect on the aroma profile.

Unrevealing the Halyomorpha halys Damage Fingerprint on Hazelnut Metabolome by Multiomic Platforms and AI-Aided Strategies.

The brown marmorated stink bug (Halyomorpha halys) poses a significant threat to hazelnut crops by affecting kernel development and causing quality defects, reducing the market value. While previous studies have identified bitter-tasting compounds in affected kernels, the impact of stink bug feeding on the hazelnut metabolome, particularly concerning aroma precursors, remains underexplored. This study aims to map the nonvolatile metabolome and volatilome of hazelnut samples obtained by caging H. halys on different cultivars in two locations to identify markers for diagnosing stink bug damage. Using a multiomic approach involving headspace solid-phase microextraction (HS-SPME), comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOF MS), and liquid chromatography-high-resolution mass spectrometry (LC-HRMS), both raw and roasted hazelnuts are analyzed, with artificial intelligence (AI) and machine learning tools employed to explore data correlations. The study finds that the hazelnut metabolome and volatilome exhibit high chemical complexity with significant classes of compounds such as aldehydes, ketones, alcohols, and terpenes identified in both raw and roasted hazelnuts. Multivariate analysis indicates that the orchard location significantly impacts the metabolome, followed by damage type, with cultivar differences being less pronounced. Partial least-squares discriminant analysis (PLS-DA) models achieve high predictive accuracy for orchard location (99%) and damage type (≈80%), with the roasted volatilome showing the highest predictive accuracy. Correlation matrices reveal significant relationships between raw hazelnut metabolites and aroma compounds in roasted samples, suggesting potential markers for stink bug damage that could guide the quality assessment and mitigation strategies. Data fusion techniques further enhance classification performance, particularly in predicting damage type, underscoring the potential of integrating multiple data sets for comprehensive quality assessment.

Characterization of Changes in Ripening Process of Volatile Apple Compounds Based on HS-SPME-GC-MS Analysis

The aim of this study was to identify the aromatic compounds present in different apple varieties and to gain insights into the changes in the aromatic compounds during ripening. Three apple varieties (“Red Astrachan”, “Ning Qiu”, and “Golden Delicious”) at different stages of ripening were selected for this study; their peel and pulp were analyzed via headspace solid-phase microextraction (HS-SPME)–gas chromatography–mass spectrometry (HS-SPME-GC-MS), and 30 volatile compounds were identified. The samples’ differences were analyzed using heat map cluster analysis, principal component analysis (PCA), and an independent samples t-test. The results showed that the content of aromatic compounds in the peel was higher than that in the pulp. The relative content of esters in the aromatic compounds of the three apple varieties followed the order of pulp &gt; peel and “Ning Qiu” &gt; “Golden Delicious” &gt; “Red Astrachan”. This suggests that “Ning Qiu” combines the advantages of its parents in terms of its aroma content. The highest concentrations of aroma compounds in “Red Astrachan” and “Ning Qiu” accumulate before the ripening stage, and care should be taken to choose an appropriate harvesting time according to the different needs during production. The main compounds of “Red Astrachan” are aldehydes and C8 esters, while those of “Ning Qiu” and “Golden Delicious” are alkenes and esters. After analyzing the relative odor activity values (rOAVs) of key volatile compounds and their aroma descriptors during the harvest period, acetic acid pentyl ester, butanoic acid hexyl ester, hexanoic acid hexyl ester, and 2-ethyl-1-Hexanol were found to contribute the most to the overall flavor of the peel and pulp. “Ning Qiu” combines the parental advantages of the concentrated peel of “Red Astrachan” and the astringent pulp of “Golden Delicious”, with compounds in its composition that give a pleasing aroma. Mature “Ning Qiu” fruits have a more intense aroma and fruity flavor. The development of flavor-specific varieties has provided the theoretical basis for future research in molecular hybridization, molecular-assisted breeding, and the molecular biology of apple flavor synthesis and metabolism.