Search Results (2,212)

A novel polysaccharide from Cynanchum auriculatum Royle ex Wight was isolated, structurally characterized, and its antioxidant activity was evaluated. The crude extract was purified by ion exchange and size exclusion chromatography to obtain a homogeneous fraction, CAP2-1. CAP2-1 displayed a weight-average molecular mass of 184.17 kDa and is mainly composed of galactose, arabinose, and galacturonic acid. Structural analysis revealed that CAP2-1 is a highly branched acidic arabinogalactan-type polysaccharide with a backbone of →6)-β-D-Galp-(1→, →3,6)-β-D-Galp-(1→, and →4)-α-D-GalpA-(1→ units, and side chains enriched in α-L-arabino furanose residues. Ultrasonic degradation produced a lower-molecular-weight derivative, UCAP2-1, which exhibited significantly stronger free radical scavenging ability compared with CAP2-1 (p < 0.01). These findings suggest that molecular weight reduction enhances antioxidant properties by improving electron-donating capacity and accessibility to reactive sites. This study reveals the structure–antioxidant relationship of CAP2-1 and UCAP2-1 and highlights UCAP2-1 as a promising natural antioxidant. Full article

Protein phosphorylation is a crucial post-translational modification that regulates protein activity, cellular signaling, transcriptional regulation, and cell cycle control. However, the analysis of phosphoproteins in biological samples is often compromised by complex sample matrices and interference from high-abundance proteins. While the top-down phosphoproteomics strategy enables comprehensive analysis of post-translational modifications based on intact proteins, its requirement for higher protein purity due to low protein ionization efficiency poses stern challenges. Consequently, developing appropriate enrichment methods for phosphoproteins in practical samples becomes essential. Immobilized metal ion affinity chromatography (IMAC) represents a common strategy for phosphorylated protein separation and enrichment. Among metal ions, Ti⁴⁺ has gained widespread application as IMAC chelating ligands due to its capacity to form multiple coordination networks and its high selectivity for phosphorylated protein enrichment, leveraging the strong chelating ability of phosphate groups toward metal ions. This paper presents the design and preparation of a novel magnetic Ti-IMAC nanocomposite, MNP@MPTMS–VPA–Ti(IV). The material is modified with phosphate groups via facile thiol-ene click chemistry and then immobilizes Ti⁴⁺, enabling selective enrichment of intact phosphoproteins through IMAC affinity. The efficiency of enrichment was evaluated using subsequent matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for detection and analysis. This Ti-IMAC material-based magnetic solid-phase extraction (MSPE)-MALDI-TOF MS protocol has been successfully applied to enrich intact phosphoproteins in milk and eel mucus with high selectivity, sensitivity, and suitability. Full article

The use of lecithin as an emulsifier in food supplements has increased in recent years. However, successful formation of liposomes or micelles requires an appropriate mixture of phospholipids in lecithin. To evaluate the emulsification properties of lecithin for food supplements, a reliable analytical procedure for characterizing phospholipids is necessary. A liquid chromatography–mass spectrometry method was developed to identify phospholipids in lecithin without standard reference materials. For efficient separation of phospholipids before mass spectrometric analysis, a reverse-phase high-performance liquid chromatography method was optimized using a Waters XBridge Protein BEH C4 column. The optimized chromatographic method demonstrated good linearity and precision. Molecular ions were detected in full scan mode to determine accurate mass-to-charge ratios for individual peaks in the chromatogram. A custom Python program was then used to generate a list of possible phospholipid species for each peak based on the measured mass-to-charge ratios. Tandem mass spectrometry was performed to confirm the identity of specific phospholipids by comparing experimental fragmentation patterns with theoretical predictions. Identification of the phospholipids was also confirmed with four commercially available standard reference compounds, demonstrating the reliability of the proposed approach. The developed method offers a practical and cost-effective strategy for identifying phospholipids in complex matrices, especially when standard reference compounds are unavailable. Additionally, it enables targeted selection of standard compounds for future quantitative analyses, making it a valuable tool for comprehensive lipid profiling. Full article

Background/Objectives: Essential amino acids’ (EAAs) biological effects depend on both gastrointestinal stability and intestinal bioavailability. A commercially available EAA blend has previously shown to be highly bioaccessible and able to inhibit the DPP-IV enzyme both directly and at a cellular level following simulated digestion in vitro. In light with this consideration, the present study aimed to evaluate the intestinal in vitro bioavailability of GAF subjected to INFOGEST digestion (iGAF) and to investigate the metabolic effects of its bioavailable fraction on muscle cells using an integrated Caco-2/C2C12 co-culture model. Methods: Differentiated Caco-2 cell lines were treated with iGAF, and amino acid transport was quantified by ion-exchange chromatography. The basolateral fraction containing bioavailable EAAs was used to treat differentiated C2C12 myotubes for 24 h. Western blot analyses were performed to assess the activation of anabolic and metabolic pathways, including mTOR, Akt, GSK3, AMPK and GLUT-4. Results: More than 50% of each EAA present in iGAF crossed the Caco-2 monolayer, with BCAAs and phenylalanine particularly enriched in the basolateral fraction. Exposure of C2C12 myotubes to the bioavailable iGAF stimulated mTORC1 activation and increased the phosphorylation of Akt and GSK3, indicating an enhanced anabolic response. At a cellular level, iGAF also elevated the p-AMPK/AMPK ratio, suggesting activation of energy-sensing pathways. Moreover, GLUT4 protein levels and glucose uptake were significantly increased. Conclusions: The study focuses exclusively on a cellular model, and results suggested that iGAF is highly bioavailable in vitro and that its absorbed fraction activates key anabolic and metabolic pathways of skeletal muscle cells, enhancing both protein synthesis signaling and glucose utilization in vitro. Full article

Hawk tea (Litsea coreana Levl. var. lanuginosa), a naturally caffeine-free herbal beverage widely consumed in Southwest China, is characterized by a pronounced camphoraceous note that often deters first-time consumers. In this study, hawk tea leaves were subjected to solid-state fermentation with four microbial strains—Monascus purpureus, Aspergillus cristatus, Bacillus subtilis, and Blastobotrys adeninivorans. The volatile compounds of unfermented and fermented hawk teas were identified by ultra-fast gas chromatography electronic nose (ultra-fast GC e-nose), gas chromatography–mass spectrometry (GC-MS) and gas chromatography–ion mobility spectrometry (GC-IMS) analyses, respectively. Furthermore, the calculation of odor activity values (OAVs) and relative odor activity value (ROAV) revealed that 6 and 25 volatile chemicals, including perillaldehyde (OAV 3.692) and linalool (ROAV 100), were the main contributors to the floral, fruity, and woody aroma of fermented hawk tea. Sensory evaluation confirmed that fermentation generally enhanced woody notes while significantly reducing the characteristic camphoraceous and oil oxidation odors. Notably, the Blastobotrys adeninivorans-fermented sample exhibited the most pronounced floral and fruity nuances, accompanied by significantly elevated aroma complexity and acceptability. Consequently, Blastobotrys adeninivorans represents a promising starter culture for the improvement of hawk tea flavor. Full article

A huge amount of waste is produced annually by the food processing industry which must be valorized into high-value products. Therefore, the aim of this work involves the use of such wastes for production of β-glucans from medicinal basidiomycete strains which are powerful biological response modifiers in several clinical disorders. The production of β-glucans from basidiomycete strains in submerged fermentation was monitored by using monoclonal antibodies of the IgG and IgM classes as well as by Congo red assay in the presence of several agro-industrial waste products such as milk permeate, waste coffee grounds, orange peels and rice husks. Subsequently, these β-glucans were purified by using gel filtration and ion-exchange chromatography. FTIR analysis of several β-glucans was carried out to investigate their structural properties. The adsorption of β-glucans on microtiter plates was dependent on the temperature as well as on the time period of immobilization for ELISA. These mAbs can be used in a competitive ELISA for detection and quantification of β-glucans from basidiomycete mushrooms. Full article

Background: Autism Spectrum Disorders (ASD) are neurodevelopmental disorders characterized by repetitive behaviors and social interaction deficits. While the severity of ASD is classified into levels (1–3) by the DSM-5, reliable circulating biomarkers to differentiate these levels are lacking. This retrospective pilot study examines plasma amino acid levels in children with ASD to identify the potential biomarkers of disease severity. Methods: Plasma samples from 30 children diagnosed with ASD (24 males, 6 females, aged 3–12 years) were analyzed. Participants were stratified into two groups based on the Autism Diagnostic Observation Schedule Calibrated Severity Score (ADOS CSS): Group 1, presenting with mild symptoms (Level 1, n = 11), and Group 2, characterized by moderate-to-severe symptoms (Levels 2–3, n = 19). This was further confirmed by the identification of electroencephalogram (EEG) anomalies (21.1%) and magnetic resonance imaging (MRI) abnormalities (5.3%), which were detected exclusively in Group 2 and absent in Group 1. Amino acid levels were measured by ion-exchange chromatography. Statistical analyses (Mann–Whitney U test and chi-square test) were used to compare AA levels between groups. Results: Statistically significant differences were observed in the levels of phosphoethanolamine, aspartic acid, and glutamic acid between the two groups. These amino acids (AA) were significantly higher in the moderate-to-severe symptoms group (Levels 2–3) compared to the mild symptoms group (Level 1) (p < 0.05). All AA values remained within age-appropriate reference ranges. Conclusions: Plasma levels of phosphoethanolamine, aspartic acid, and glutamic acid may serve as potential biomarkers for ASD severity in children. Results from this exploratory analysis suggest that AA profiling could differentiate ASD severity and identify specific metabolic pathways, such as excitatory neurotransmission and phospholipid turnover. Further studies with larger cohorts are necessary to validate these findings and explore the role of AAs in ASD pathophysiology. Full article

Background: Uncontrolled diabetes is characterised by a loss of blood glucose control and increased oxidation of fatty acids to produce ATP. Use of metabolic inhibitors to blunt fatty acid oxidation and restore glucose metabolism is a poorly studied intervention for diabetes. Methods: Steptozotocin-induced diabetes was developed in Wistar male rats. A subset was supplemented with mildronate (100 mg/kg—14 days). Exploiting liquid chromatography-mass spectrometry for workflows including ion exchange-, C18-reverse phase- and HILIC-based chromatography methods, metabolite levels were quantified in plasma liver and brain tissue. Using both untargeted and targeted metabolomic analysis changes to the global tissue metabolome and individual metabolic pathways were estimated. Results: We document that an inhibitor of carnitine synthesis, mildronate, decreased plasma (50% p < 0.01) carnitine abundance and decreased plasma glucose concentration by one-third compared to streptozotocin (STZ)-treated rats (p < 0.001). Targeted metabolomic analysis of the liver showed decreased alpha-ketoglutarate abundance (35% p < 0.05) by STZ diabetes that was further decreased following mildronate treatment (50% p < 0.05). For both beta-hydroxybutyrate and succinate levels, STZ diabetes increased hepatic abundance by 50% (p < 0.05 for both), which was restored to control levels by mildronate (p < 0.05 for both). In contrast, brain TCA intermediate abundances were unaffected by either STZ diabetes or mildronate (NS for all). STZ diabetes also decreased abundance of pentose phosphate pathway (PPP) metabolites in the liver (glucose-6-phosphate, 6-phosphogluconolactone, 6-phosphogluconate 50% for all; p < 0.05), which was not restored by mildronate treatment. However, brain PPP metabolite abundance was unchanged by STZ diabetes or mildronate (NS for all). However, mildronate treatment did not affect the increased abundance of brain sorbitol, sorbitol-6-phosphate and glucose-6-phosphate as a result of STZ diabetes. Conclusions: Together, these observations highlight the potential role that metabolic inhibitors, like mildronate, may play in restoring blood glucose for diabetic patients, without a direct effect of tissues that represent obligate consumers of glucose (e.g., brain) whilst manipulating fat oxidation in tissues such as the liver. Full article

This study investigated the effect of superheated steam (SS) assisted glycosylation modification on the flavor profile of oyster peptides (OP), and explored the correlation between key flavor compounds and glycosylation degree using Gas Chromatography–Ion Mobility Spectrometry (GC-IMS) and nano-scale Liquid Chromatography coupled with High-Resolution Mass Spectrometry (nano-LC/HRMS). The results indicated that SS treatment accelerated the glycosylation process, reduced free amino groups level, and distinguished their unique flavor through E-nose. GC-IMS analysis detected 64 signal peaks including 13 aldehydes, 6 ketones, 7 esters, 6 alcohols, 2 acids, 2 furans and 5 other substances. And it was revealed that SS-mediated glycosylation treatment reduced the levels of fishy odorants like Heptanal and Nonanal, while promoting the pleasant-smelling alcohols and esters. In addition, Pearson correlation showed a positive correlation between excessive glycation and the increase in aldehydes, which might cause the recurrence of undesirable fishy notes. Further nano-LC/HRMS analysis revealed that arginine and lysine acted as the main sites for glycosylation modification. Notably, glycosylated peptides such as KAFGHENEALVRK, DSRAATSPGELGVTIEGPKE, generated by mild SS treatment could convert into ketones and pyrazines in subsequent reactions, thereby contributing to overall sensory enhancement. In conclusion, SS treatment at 110 °C for 1 min significantly improved the flavor quality of OP and sustains improvement in subsequent stages, providing theoretical support for flavor optimization of oyster peptides. Full article

Volatile fingerprint analysis using Gas Chromatography–Ion Mobility Spectrometry (GC-IMS) was applied to differentiate cowhide (22 samples), sheepskin (6 samples), and pigskin (6 samples). A total of 126 signal peaks were detected from the whole GC-IMS dataset, with 96 volatile compounds identified. Principal Component Analysis (PCA) revealed distinct clustering: cowhide exhibited unique volatile profiles, separating itself clearly from sheepskin and pigskin, which showed significant similarity. This was confirmed by Hierarchical clustering, K-means clustering (optimal k = 2), and Partial Least Squares Discriminant Analysis (PLS-DA) (R² = 0.9836, Q² = 0.9040). Cowhide was characterized by exclusive compounds (2-Hexanone, alpha-Thujene, Butyl acetate, 3-Methyl-2-butanol, 2-Heptanone, Hexyl methyl ether-monomer, Diethyl disulfide). Sheepskin and pigskin shared exclusive compounds (2-Methyl propanol, Isobutyl acetate, 2-Pentyl acetate, 3-Penten-2-one, 2,5-Dimethylfuran). Orthogonal PLS-DA (OPLS-DA) further differentiated sheepskin (Ethyl isobutanoate-dimer, Pentyl acetate-dimer, 3-Methyl-2-butanol, 2-Pentanone, 2-Methylbutanol-dimer, 3-Methyl-1-butanol, 2,5-Dimethylfuran, Propan-2-ol, Ethanol-dimer, and alpha-Thujene) and pigskin (Butan-2-one, Pentanal-dimer, 1-Pentanal-monomer, Ethyl vinyl ether, Z-2-Heptene, and Butyronitrile), identifying alpha-Thujene, 3-Methyl-2-butanol, and 2,5-Dimethylfuran as universal discriminatory markers. GC-IMS coupled with chemometric analysis provides a robust approach for leather authentication. Full article

This study aimed to analyze the volatile organic compounds (VOCs) for different rice aroma types using sensory evaluation, headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS), and gas chromatography-ion mobility spectrometry (GC-IMS) techniques, and to explore the material basis for the flavor differences. Based on the sensory evaluation results, rice aroma was categorized into three types, distinguished by their unique aroma compounds. Type A was characterized by a prominent sweet, popcorn aroma, type B by a more prominent cereal and starchy flavor, and type C by a more complex aroma. Untargeted metabolomics analysis using HS-SPME-GC-MS identified and characterized 74 volatile compounds. A comparison of A versus B versus C revealed 8 key aroma compounds, primarily alkanes, aldehydes, ketones, alcohols, and heterocyclic compounds. (E)-2-Octenal, 6-Undecanone, 2-Acetyl-1h-pyrrole, and P-menthan-1-ol in type A gave it a better sweet aroma, Dodecane, 2,6,10-trimethyl-, 1-Octen-3-one, and 2-Methyldecane in type B gave it a better starchy and cereal flavor. 2-Acetyl-1h-pyrrole, Heptacosane, and 1-Propanol in type C contributed to a complex aroma. GC-IMS analysis showed that the fingerprints of rice with different aroma types were significantly different. The VOCs of aroma type A contained (+)-limonene, 2-methylpyrazine, 2-pentanone, ethyl butanoate, n-pentanal, styrene, 1-butanol, 3-methyl-, acetate, 1-hexanal, 1-pentanol, and 2-heptanone, which gave it a better sweet aroma. The VOCs of aroma type C contained 1-octen-3-ol, 2,6-dimethyl pyrazine, 2-acetylpyridine, and ethyl hexanoate, which gave it a better complex aroma. Full article

This study investigated the site-specific ionic composition and wet deposition loads of rainwater collected from eight actively cultivated agricultural regions across South Korea, with the aim of quantifying spatial and seasonal variability and interpreting how regional agricultural characteristics and surrounding site conditions influence major ion concentrations and deposition patterns. Rainfall samples were obtained using automated samplers and analyzed via high-performance ion chromatography for major cations (Na⁺, $\mathrm{N}{\mathrm{H}}_4^{+}$, K⁺, Ca²⁺, Mg²⁺) and anions (Cl⁻, $\mathrm{N}{\mathrm{O}}_3^{-}$, $\mathrm{S}{\mathrm{O}}_4^{2-}$, $\mathrm{N}{\mathrm{O}}_2^{-}$). The results revealed significant seasonal fluctuations in ion loads, with $\mathrm{N}{\mathrm{H}}_4^{+}$ (peak 1.13 kg/ha) and K⁺ (peak 0.25 kg/ha) reaching their highest levels during summer due to increased fertilizer use and crop activity. Conversely, Cl⁻ peaked in winter (2.11 kg/ha in December), particularly in coastal regions, likely influenced by de-icing salts and sea-salt aerosols. Correlation analysis showed a strong positive association among $\mathrm{N}{\mathrm{H}}_4^{+}$, $\mathrm{N}{\mathrm{O}}_3^{-}$, and $\mathrm{S}{\mathrm{O}}_4^{2-}$ (r = 0.89 and r = 0.84, respectively), indicating shared atmospheric transformation pathways from agricultural emissions. Ternary diagram analysis further revealed regional distinctions: coastal regions such as Gimhae and Muan exhibited Na⁺ and Cl⁻ dominance, while inland areas like Danyang and Hongcheon showed higher proportions of Ca²⁺ and Mg²⁺, reflecting differences in aerosol sources, land use, and local meteorological conditions. These findings underscore the complex interactions between agricultural practices, atmospheric processes, and local geography in shaping rainwater chemistry. The study provides quantitative baseline data for evaluating non-point source pollution and developing region-specific nutrient and soil management strategies in agricultural ecosystems. Full article

The flavor and aroma of kiwifruit are largely influenced by the concentration of Volatile Organic Compounds (VOCs). To analyze the volatile profiles and identify characteristic aroma compounds, this study utilized Gas Chromatography-Ion Mobility Spectrometry (GC-IMS) to analyze the aromatic compounds sourced from seven major production regions in China and New Zealand, covering red-, green-, and yellow-fleshed varieties. A total of 77 VOCs were identified, with esters, aldehydes, and ketones as the dominant classes. Significant regional and varietal differences were observed: red-fleshed kiwifruits from Yunnan exhibited high levels of 2-Vinyl-5-methylfuran, Ethyl formate, and 1-Penten-3-one; green-fleshed fruits from Shaanxi were rich in Limonene and Methyl hexanoate, and those from Yunnan were rich in 1-Propanol and 1-Hexanol; and yellow-fleshed fruits from Henan were characterized by Methyl salicylate and 3-Hydroxy-2-butanone. Orthogonal partial least squares discriminant analysis (OPLS-DA) successfully classified kiwifruits by origin and variety, confirming the stability and predictive power of the model (Q2Y > 0.97). This study also elucidated the key metabolic pathways—including lipid oxidation, amino acid degradation, and terpenoid metabolism—underlying the formation of these characteristic VOCs. These findings provide a theoretical foundation for the biochemical regulation of kiwifruit flavor and support the development of origin-tracing and quality-assessment tools based on VOC fingerprints. Full article

This study aimed to characterize the physicochemical, structural, and volatile compound changes in commercially sterilized pre-cooked braised chicken (PBC) during storage at 25 °C, using analyses conducted every 30 days from 30 to 180 days. Assessed parameters included microstructure, color, texture, pH, malondialdehyde (MDA) content, Ca²⁺-ATPase activity, and volatile organic compounds (VOCs). Significant quality changes occurred during storage. Specifically, the L* value decreased, and the a* value increased, while hardness, springiness, chewiness, and Ca²⁺-ATPase activity declined. pH increased from 6.01 to 6.59, and MDA content rose from 10.16 to 23.42 nmol/g. 91 VOCs were identified by gas chromatography-ion mobility spectrometry (GC-IMS), comprising 13 alcohols, 18 aldehydes, 18 ketones, 3 acids, 9 esters, 12 hydrocarbons, 6 aromatics, and 12 others. VOC profiles shifted dynamically: key aldehydes and ketones decreased initially, then increased, whereas alcohols, esters, hydrocarbons, and sulfur-containing compounds increased, then decreased. Prolonged storage significantly deteriorated the quality and altered the flavor profile, providing insights for PBC storage. Full article

Background/Objectives: Given the clinical limitations of azathioprine (AZA) in treating inflammatory bowel disease, this study developed an AZA-loaded microbiota-modulating and colon-targeted nanoparticle constructed from pectin, Zein, and Eudragit^®S100 (APZE), which was hypothesized to enhance efficacy while reducing toxicity. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established to simultaneously quantify AZA and its metabolites, enabling the investigation of the pharmacokinetic and microbial metabolism differences between APZE and AZA suspension (AZAS). Methods: APZE was characterized, and an LC-MS/MS method was developed for quantifying AZA and its metabolites in multiple matrices. Given the potential of APZE for colon targeting and modulation of the microbiota, which may affect drug absorption, distribution, and microbiota-mediated metabolism, we determined analyte concentrations in rat plasma, tissues, and microbial cultures at different time points following administration of APZE or AZAS. Results: AZA, 6-mercaptopurine (6-MP), 6-methylmercaptopurine (6-MMP), and 6-thioguanine (6-TG) were quantified in positive ion mode, and 6-thiouric acid (6-TU) in negative ion mode. The assay demonstrated excellent accuracy, precision, and stability over the concentration range of 5–1000 ng/mL. Orally administered APZE exhibited higher bioavailability, improved intestinal absorption, and reduced formation of the inactive metabolite 6-TU compared to AZAS. In microbial cultures, AZA was metabolized primarily to 6-MP, and APZE underwent more extensive metabolism to 6-MP than AZAS. Conclusions: This method provides accurate and precise quantification of physiologically relevant concentrations of AZA and its metabolites (6-MP, 6-MMP, 6-TG, and 6-TU), offering a bioanalytical tool for the pharmacokinetic and gut microbiota metabolism studies of AZA formulations. These findings suggest that APZE is a promising drug delivery formulation. Full article