Liquid Mass Spectrometry Research Articles

Dry crude pomegranate peel extract as a bio-input for medicinal pharmaceutical gel with healing activity.

The sustainable use of pomegranate peel, a by-product of the food industry, is gaining importance in developing pharmaceutical bio-inputs, aligning with circular economy practices and waste reduction. This study explores the application of dry crude pomegranate peel extract (PPE) as a bio-input for medicinal gels with wound healing properties. PPE was extracted via percolation in ethanol and freeze-dried. High-performance liquid chromatography (HPLC) and mass spectrometry identified key bioactive compounds: gallic acid, punicalagin, punicalin, ellagic acid, and citric acid, recognized for their antioxidant, antimicrobial, and healing properties. In vitro assays revealed low hemolytic activity, non-cytotoxicity, significant fibroblast proliferation, and robust antioxidant activity (95.99% DPPH scavenging at 100 µg/mL). A carbomer gel containing 2.5% w/w of the extract effectively healed wounds in rats, with performance comparable to silver sulfadiazine ointment, while demonstrating no microbial contamination during the process. These findings position PPE as a promising, sustainable, and effective alternative for wound care pharmacotherapy, addressing both medical needs and environmental sustainability.

Revealing New Analytical Insights into RNA Complexes: Divalent siRNA Characterization by Liquid Chromatography and Mass Spectrometry.

Accurate characterization of therapeutic RNA, including purity and identity, is critical in drug discovery and development. Here, we utilize denaturing and non-denaturing chromatography for the analysis of ∼25 kDa divalent small interfering RNA (di-siRNA), which comprises a complex 2:1 triplex structure. Ion pair reversed-phase (IPRP) liquid chromatography (LC) experiments with UV absorbance and mass spectrometry (MS) showcase a single denaturing LC method for identity confirmation, impurity profiling, and sequencing with automated MS data interpretation. IPRP, size exclusion chromatography (SEC), and melting temperature (Tm) experiments showcase the need for consideration of chromatographic conditions in evaluating noncovalent siRNA structures─here, low-temperature IPRP experiments (generally considered "non-denaturing") indicate denaturation of the noncovalent complex for certain di-siRNA sequences, while SEC data indicate that di-siRNA aggregation can be vastly underestimated due to sample dilution prior to LC experiments. Furthermore, SEC data critically show the propensity of denatured di-siRNA samples to renature under SEC mobile phase conditions upon exposure to high ionic/salt solutions, corroborated by Tm experiments. This work highlights the need for consideration of the noncovalent nature of certain RNA therapeutics during sample preparation, method development, and analytical characterization and the often sequence-specific strength of complex formation, which may significantly affect analytical results obtained for such molecules. Attention to the highly dynamic nature of the duplex RNA structure, which is heavily influenced by its environment, must be considered during analytical method development.

Recent advances in electrochemical sensing and remediation technologies for ciprofloxacin.

Ciprofloxacin (CIP) is an extensively used broad-spectrum, fluoroquinolone antibiotic used for treating diverse bacterial infections. Effluent treatment plants (ETPs) worldwide lack technologies to detect or remediate antibiotics. CIP reaches the aquatic phase primarily due to inappropriate disposal practices, lack of point-of-use sensing, and preloaded activated charcoal filter at ETPs. The co-existence of bacteria and CIP in such aqueous pools has promoted fluoroquinolone resistance in bacteria and should be minimized. The worldwide accepted standard detection methodologies for the detection of CIP are high-performance liquid chromatography and mass spectrometry, which are lab-based, require state-of-the-art equipment, and are expensive. Hence, it is difficult to integrate them for on-site monitoring. Further, the current remediation technologies like conventional sludge-treatment techniques fail to remove antibiotics such as CIP. Several point-of-use technologies for the detection of CIP are being investigated. These typically involve the development of electrochemical sensors where substrates, modifiers, biorecognition elements, and their chemistries are designed and optimized to enable robust, point-of-use detection of CIP. Similarly, remediation techniques like adsorption, membrane filtration, ion exchange, photocatalysis, ozonation, oxidation by Fenton's reagent, and bioremediation are explored, but their onsite use is limited. The use of these sensing and remediation technologies in tandem is possibly the only way the issues related to antimicrobial resistance may be effectively tackled. This article provides a focused critical review on the recent advances in the development of such technologies, laying out the prospects and perspectives of their synergistic use to curb the menace of AMR and preserve antibiotics.

Simultaneous Profiling of Multiple Phosphorylated Metabolites in Typical Biological Matrices via Ion-Pair Reversed-Phase Ultrahigh-Performance Liquid Chromatography and Mass Spectrometry.

Simultaneous analysis of multiple phosphorylated metabolites (phosphorylated metabolome) in biological samples is vital to reveal their physiological and pathophysiological functions, which is extremely challenging due to their low abundance in some biological matrices, high hydrophilicity, and poor chromatographic behavior. Here, we developed a new method with ion-pair reversed-phase ultrahigh-performance liquid chromatography and mass spectrometry using BEH C18 columns modified with hybrid surface technology. This method demonstrated good performances for various phosphorylated metabolites, including phosphorylated sugars and amino acids, nucleotides, NAD-based cofactors, and acyl-CoAs in a single run using standard LC systems. Specifically, the method showed good retention (capacity factor > 2) and reproducibility (ΔtR < 0.09 min, n = 6), peak symmetry (tailing factor < 2), and sensitivity (limit-of-detection < 238 fmol-on-column with QTOFMS) for all tested analytes especially for the medium- and/or long-chain acyl-CoAs. The method demonstrated reproducible applicability across numerous biological matrices, including tissue (liver), human biofluids (urine, plasma), cells, and feces, and revealed significant molecular phenotypic differences in phosphorylated metabolite composition.

Unlocking Nutraceutical Potential: Metabolomic Analysis Reveals Therapeutic Compounds in Rice

Background Rice varieties enriched with potential bioactive compounds are poised to make a significant contribution to global health. It is anticipated that edible medications made from rice seeds will lead to novel therapeutics. Purpose High-yielding popular pigmented and nonpigmented rice varieties were selected for in-depth analysis of bioactive contents. Nutraceutical potential of high-yielding rice varieties often goes underexplored compared to low-yielding specialty rice varieties. This study addresses the gap by metabolomic profiling of popular rice varieties to uncover bioactive compounds with significant pharmaceutical and nutritional benefits. Methods Untargeted metabolomic profiling of the brown rice of pigmented and nonpigmented varieties was conducted by Orbitrap-high-resolution liquid chromatography and mass spectrometry, and primary and secondary metabolites were identified and annotated. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging (RS) assay was carried out to identify the variety with superior antioxidant activity. Correlation analysis by using Metaboanalyst software identified major metabolites with RS potential. Over-representation analysis was performed for the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and major pathways were identified for metabolites with nutraceutical potential in pigmented rice. Results Major metabolites with antioxidant, anti-inflammatory, and anticarcinogenic properties were identified from rice varieties. Bioactive compounds detected from brown rice included choline, didodecyl-3,3-thiodipropionate, conjugated linoleic acid, oleic acid, 13( S)-HOTrE, monoolein, nicotinic acid, pantothenic acid, isoquinoline, trigonelline, corymboside, ferulic acid, and 2,4-quinolinediol. The DPPH RS assay revealed superior antioxidant activity in the pigmented rice, with differential metabolites from the pathways of phenylpropanoid biosynthesis, arginine and proline metabolism, and biosynthesis of unsaturated fatty acids showing a significant correlation ( p &lt; .01) with RS potential. Conclusion Catechin, (–)-epicatechin, (±)12(13)-DiHOME, 4-guanidinobutyric acid, and 5-methoxysalicylic acid are key metabolites that emphasize the antioxidant potential of red-colored rice as a functional food with significant health benefits. The study highlights the therapeutic promise of pigmented whole-grain rice.

In vitro analysis of composition profiles of resins for 3D printing of dentures.

This study aimed to investigate the resin compounds from CAD-CAM 3D-printed denture resins, focusing on the identification and classification of free monomers and other components. The primary objective was to determine the chemical profile of these 3D-prinding resin materials. Four 3D-printed denture resins, two base materials (1: DentaBASE, Asiga Ltd., Alexandria, NSW, Australia; 2: DENTURETEC, SAREMCO Dental AG), and two tooth-colored materials (1: DentaTOOTH, Asiga Ltd.; 2: CROWNTEC (SAREMCO Dental AG), were analyzed using high-performance liquid chromatography and mass spectrometry (HPLC-MS). Data analysis was processed including peak alignment and normalization to an ethanol blank, resulting in detailed identification of compounds in the resin. Out of 5,208 detected compounds in the resin, 63 were retained after applying filtering criteria for further analysis, categorized into methyl methacrylate (MMA) and derivatives (31), photo initiators (8), UV stabilizers (1), and other additives (23). Sample 2B exhibited the highest number of resin compounds (62), including the most MMA derivatives and additives. MMA accounted for over 69% in abundance across samples, with unique additives such as ethyl-4-dimethylaminobenzoate and bis(acryloyloxymethyl) tricyclo [5.2.1.02,6] decane (TCD-DI-HEA) identified. Variations in the resin composition distribution and abundance highlighted differences in resin formulations. Within the limits of this in vitro, exploratory study, unpolymerized 3D-printed denture resins were analyzed, and various resin compounds in 3D-printed denture resins, including MMA, photo initiators, UV stabilizers, and additives, were identified along with significant variability between samples. While 3D-printing enhances efficiency and customization, further investigation is needed to better understand and assess the long-term intraoral effects and their effectiveness in complete dentures. This study provides insights into the chemical composition of 3D-printed denture resins, highlighting variability in resin compound profiles. Understanding these materials is essential for ensuring biocompatibility and performance. The findings support the need for further research and standardized testing to guide clinicians in the safe and effective use of 3D-printed dentures.

Toward Resolving Heterogeneous Mixtures of Nanocarriers in Drug Delivery Systems through Light Scattering and Machine Learning.

Nanocarriers (NCs) have emerged as a revolutionary approach in targeted drug delivery, promising to enhance drug efficacy and reduce toxicity through precise targeting and controlled release mechanisms. Despite their potential, the clinical adoption of NCs is hindered by challenges in their physicochemical characterization, essential for ensuring drug safety, efficacy, and quality control. Traditional characterization methods, such as dynamic light scattering and nanoparticle tracking analysis, offer limited insights, primarily focusing on particle size and concentration, while techniques like high-performance liquid chromatography and mass spectrometry are hampered by extensive sample preparation, high costs, and potential sample degradation. Addressing these limitations, this work presents a cost-effective methodology leveraging light scattering and optical forces, combined with machine learning algorithms, to characterize polydisperse nanoparticle mixtures, including lipid-based NCs. We prove that our approach provides quantification of the relative concentration of complex nanoparticle suspensions by detecting changes in refractive index and polydispersity without extensive sample preparation or destruction, offering a high-throughput solution for NC characterization in drug delivery systems. Experimental validation demonstrates the method's efficacy in characterizing commercially available synthetic nanoparticles and Doxoves, a liposomal formulation of Doxorubicin used in cancer treatment, marking a significant advancement toward reliable, noninvasive characterization techniques that can accelerate the clinical translation of nanocarrier-based therapeutics.

Optimization of liquid chromatography and mass spectrometry parameters based on LC-QQQ: A case study on lysinoalanine.

Lysinoalanine (LAL), commonly formed in high-protein foods, raises concerns due to its nephrotoxicity and potential reduction in nutritional properties, making its accurate detection crucial for food safety. Liquid chromatography-tandem quadrupole mass spectrometry (LC-QQQ) plays a pivotal role in the quantification of compounds, and its accuracy and sensitivity are significantly influenced by specific liquid chromatography (LC) and mass spectrometry (MS) parameters. However, the procedure and considerations for LC and MS parameters optimization have often not been discussed in depth in existing literature. Therefore, this study used LAL as a model compound to systematically optimize the key LC and MS parameters using LC-QQQ. The optimized MS parameters were as follows: parent ion m/z-234.2, capillary voltage-3.5kV, cone voltage-30V, desolvation temperature-500°C, daughter ion m/z-84.2, and collision voltage-20V. The optimized LC parameters were as follows: buffer-0.1% formic acid (v/v), column, Polaris 3 Amide-C18 (150×3 mm, 3 μm). Under these optimized conditions, the limit of detection (LOD) for LAL was detected as 13ng/mL in multiple reaction monitoring mode, which is considerably lower than the 125ng/mL detected by LC-QQQ and marginally lower than the 15.23ng/mL detected by LC-quadrupole Exactive Orbitrap MS reported in previous studies. Additionally, this study elucidates the critical factors to be considered when selecting LC and MS parameters, providing valuable insights into the detection of other compounds using LC-QQQ.

Enzymatic hydrolysis-assisted magnetic beads adsorption for precise localization of IgE-linear epitopes in ovalbumin and the mechanism of reduced allergenicity after enzymatic hydrolysis

We present a method for precisely localizing IgE-linear epitopes by adsorption of peptides containing IgE-linear epitopes onto magnetic beads. Optimal enzymatic hydrolysis conditions (55 °C, 40 min) were determined to disrupt the OVA structure, facilitating specific adsorption of peptides containing IgE-linear epitopes onto magnetic beads. Using Liquid-mass spectrometry, we identified fourteen peptides with IgE-linear epitopes, among which epitopes 3 (T91-F99), 4 (L124-F134), 6 (V160-D167), 9 (V243-E248), and 13 (S317-K322) were found to be high-frequency IgE-linear epitopes. Additionally, epitopes 1 (V41-Q52), 3 (T91-F99), 8 (Y212-A220), 11 (N292-V296), and 13 (S317-K322) were identified as previously unreported IgE-linear epitopes. Through solid-phase peptide synthesis, indirect competitive ELISA and molecular docking. The results showed that the above five peptides were mainly bound to MHC-II through hydrogen bonding. Among them, epitope 3 has the strongest affinity for MHC-II. In conclusion, magnetic beads adsorption of peptides containing IgE-linear epitopes was a novel method for the precise localization of IgE-linear epitopes. This technique was facile to execute and enables rapid and accurate identification of IgE-antigen binding sites.

Separation, Identification, and Characterization of Transformation Products Related to Relugolix Upon Subjecting to In Vitro Gastrointestinal Fluids Utilizing Liquid Chromatography, Mass Spectrometry, and Nuclear Magnetic Resonance Spectroscopy

ABSTRACTThis study investigated the gastrointestinal stability of relugolix, a biopharmaceutical classification system Class IV drug. Relugolix was found to be unstable in simulated gastric fluid but stable in simulated intestinal fluid and fed‐state–simulated gastric fluid. The high‐performance liquid chromatography method was developed and validated for separating the drug and its transformation products. The optimized method comprises mobile phase ammonium acetate (pH 4.75) as Solvent A and acetonitrile as Solvent B. A Phenomenex Gemini C18 (250 × 4.6 mm, 5 µm) column was used as the stationary phase. The transformation product, desmethyl relugolix, was identified and characterized using high‐resolution mass spectrometric and nuclear magnetic spectroscopic studies by enriching the compound using forced degradation studies under acidic stress hydrolytic conditions. Sodium alginate microspheres were prepared and evaluated to improve relugolix stability in the stomach. Additionally, gastrointestinal stability studies demonstrated that the microspheres protected relugolix from degradation in simulated gastric fluid. In silico analysis predicted minimal toxicity risks for both relugolix and desmethyl relugolix. These findings suggest that relugolix sodium alginate microspheres could be a promising strategy to mitigate the formation of desmethyl relugolix by protecting it from degradation in the stomach's acidic environment.

In vitro simulated digestion and Caco-2 cell absorption to explore potential of soy protein isolates as whey protein substitutes in canine and feline food.

Digestive properties and bioavailability of soy protein isolate and whey protein in canine and feline were evaluated using in vitro simulated digestion and cell absorption experiments. The amino acids and peptides in soybean protein isolate and whey protein after digestion were detected by high-performance liquid chromatography and liquid mass spectrometry. Gastrointestinal digestion rates of soy protein isolate and whey protein were analyzed by fitting digestive kinetics curves. The degree of absorption and utilization of amino acids and peptides from the digestive products was evaluated by establishing a Caco-2 cell model. The results demonstrated that soy protein isolate, like whey protein, could produce large amounts of free amino acids and peptides distributed 100-1500 Da. The gastrointestinal digestion speed of soy protein isolate was similar with whey protein. It is worth noting from the intestinal absorption of Caco-2 cell that more than 15 amino acids produced by soy protein isolate and whey protein could be absorbed, and a large number of peptides were also utilized by Caco-2 cell. The bioavailability of soy protein isolates and whey protein for dogs respectively reached 6.30% and 9.00%, and it reached 7.40% and 16.40% in cats, respectively. Soy isolate proteins can be digested and absorbed by pets like whey proteins. These findings may provide significant strategies and support for the application of plant-based proteins in pet foods.

The Association Between Serum Trimethylamine N-Oxide and Arterial Stiffness in Chronic Peritoneal Dialysis Patients: A Cross-Sectional Study.

Trimethylamine N-oxide (TMAO), a gut microbiome-derived metabolite, participates in the atherogenesis and vascular stiffening that is closely linked with cardiovascular (CV) complications and related deaths in individuals with kidney failure undergoing peritoneal dialysis (PD) therapy. In these patients, arterial stiffness (AS) is also an indicator of adverse CV outcomes. This study assessed the correlation between serum TMAO concentration quantified with high-performance liquid chromatography and mass spectrometry and central AS measured by carotid-femoral pulse wave velocity (cfPWV) in patients with chronic PD. Of the 160 participants included, 23.8% had a cfPWV of ≥10 m/s, which fulfilled the AS criteria. Multivariable logistic regression analysis revealed that TMAO, age, and waist circumference were positively associated with AS. Multivariable stepwise linear regression showed that underlying diabetes, advanced age, waist circumference, systolic blood pressure, and logarithmic-transformed TMAO were independently correlated with cfPWV. The area under the receiver operating characteristic curve for TMAO in differentiating AS from non-AS was 0.737. In conclusion, serum TMAO level was significantly independently correlated with central AS among participants undergoing PD for end-stage kidney failure.

Pre- and postpartum metabolizable protein supply: II. Effects on plasma amino acids and markers of tissue mobilization in transition Holstein dairy cows.

The influence of diet composition on the degree of adipose and lean muscle mobilization and concentrations of circulating AA has been demonstrated during the transition period. Altering the MP supply might offer a strategy to control tissue mobilization and increase circulating AA availability, but the optimum supply of MP fed pre- and postpartum remains unknown. We investigated the effect of increasing the MP supply in the prepartum, postpartum, or both diets on plasma AA concentrations and ultrasound and circulating indicators of tissue mobilization. Multiparous Holstein cows (n = 96) were assigned to 1 of 4 treatment groups at 28 d before expected calving following a randomized block design. Prepartum diets were formulated to contain either a control (C; 85 g of MP/kg DM; 1,175 g of MP/d) or high (H; 113 g of MP/kg DM; 1,603 g of MP/d) level of estimated MP. From calving to 21 DIM, fresh diets were formulated to contain either a control (C; 104 g of MP/kg DM; 2,044 g of MP/d) or high (H; 131 g of MP/kg DM; 2,685 g of MP/d) level of estimated MP. To control the potential confounding effect of Met and Lys supply, diets were formulated to supply an equal amount at 1.24 and 3.84 g/Mcal of ME in both prepartum diets and 1.15 and 3.16 g/Mcal of ME in both postpartum diets, respectively. The combination of a pre- and postpartum diet resulted in 4 treatment groups: 1) CC (n = 23), 2) CH (n = 24), 3) HC (n = 22), and 4) HH (n = 23). A common lactation diet (113 g of MP/kg DM; 2,956 g of MP/d) was fed from 22 DIM to the end of the observation period at 42 DIM. Transcutaneous ultrasonography was used to determine the longissimus dorsi muscle diameter and backfat thickness. Concentrations of plasma AA, 3-methylhistidine (3MH), and creatinine were determined on a subset of cows (n = 60) using ultra-high-performance liquid chromatography and mass spectrometry. Treatment did not affect the longissimus dorsi muscle diameter from -14 to 21 d relative to calving, but the diameter was greater in CH compared with HH at 40 DIM. Backfat thickness and the ratio of 3MH to creatinine did not differ by treatment. Concentrations of EAA were greater at -13 d relative to calving in HH compared with CC and CH and at -6 d relative to calving EAA concentrations were higher in HC compared with CC. Cows fed the H diet postpartum had elevated EAA concentrations at 6 and 20 DIM compared with cows fed the C postpartum diet but EAA concentration did not differ at 40 DIM. Total NEAA concentrations were higher in CH compared with HC and HH at -6 d relative to calving, but NEAA concentration did not differ by treatment at -13, 6, 20, or 40 d relative to calving. In conclusion, increasing the supply of MP fed prepartum, postpartum, or both had minimal effects on tissue mobilization but influenced concentrations of plasma AA.

Open-source Electrophilic Fragment Screening Platform to Identify Chemical Starting Points for UCHL1 Covalent Inhibitors

Target-based screening of covalent fragment libraries with mass spectrometry has emerged as a powerful strategy to identify chemical starting points for small molecule inhibitors or find new binding pockets on proteins of interest. These libraries span diverse chemical space with a modest number of compounds. Screening covalent fragments against purified protein targets reduces the demands on the mass spectrometer with respect to absolute throughput, detection limit, and dynamic range. Given these relaxed analytical requirements, we sought to develop an open-source, medium-throughput mass spectrometry system for target-based covalent fragment screening. Our platform comprises automated, dual LC desalting columns integrated with electrospray ionization for rapid sample introduction and mass spectrometry detection. The system is operated through a simple python graphical user interface running on commodity microcontroller boards which allow integration with diverse liquid chromatography and mass spectrometry instruments. We provide scripts for fragment pooling, construction of sample batches, along with routines for data processing and visualization. The system enables primary screening of ∼10,000 covalent fragments per day in pooled format. In a proof-of-concept study we executed primary and secondary screens to identify 27 hit fragments against UCHL1, a deubiquitinating enzyme that is emerging as a drug target of interest across multiple clinical indications. We validated and triaged these covalent compounds through a series of orthogonal biochemical and chemoproteomic assays. The most promising chloroacetamide covalent fragment inhibited UCHL1 activity in vitro (IC50 <5 µM) and exhibited dose-dependent binding along with good selectivity against 57 cellular DUBs as quantified by activity-based protein profiling.

Construction of Lamellar CoFe-LDHs@MoS2 to Promote Permonosulfate Properties Leading to Effective Photocatalytic Degradation of Norfloxacin

The utilization of the photo catalytic activation of permonosulfate (PMS) for the combined breakdown of pollutants has become a focal point in research. Layered double hydroxides (LDHs) have a unique layered structure which is conducive to the adsorption and diffusion of reactants, and can provide more active sites for photocatalytic reactions. The anions between the layers can be exchanged with a variety of substances so that specific catalytically active species can be introduced as needed. LDHs themselves have certain catalytic activity, which can produce synergistic catalysis between LDHs and the supported photocatalytic active substances, and further improve the degradation effect of antibiotics. In actual wastewater treatment, LDHs as a catalyst carrier have a good application prospect. However, the poor activation effect is attributed to the low separation efficiency of catalyst carriers and insufficient active sites. In this study, a dual active site system consisting of Co and Fe, known as CoFe-LDHs@MoS2, was developed as a catalyst to facilitate the synergistic degradation of norfloxacin (NOF) by PMS under visible light. The findings demonstrate that the material possesses an effective capacity for the synergistic degradation of NOF. A comprehensive investigation was conducted to assess the impact of different catalysts, PMS dosage, degradation systems (Vis, PMS, or Vis PMS), catalyst dosage, NOF concentration, pH, and cycle times on the degradation performance. The active free radicals, degradation pathways, and intermediate toxicity were elucidated through capture experiments, Electron Paramagnetic Resonance Spectrometer (ESR) analysis, a liquid mass spectrometry (LC-MS) toxicity assessment, and theoretical calculations. This research offers a novel approach for designing catalysts with exposed high activity sites for the effective removal of NOF from environmental water.

Extraction and Identification of Flavonoids from the Leaves of Pilocarpus microphyllus: Focus on Antioxidant Activity and Neuroprotective Profile

This work is based on research aiming to extract and identify flavonoids from jaborandi (Pilocarpus microphyllus) leaves and investigate their antioxidant and acute antinociceptive capacity. Characterization of the constituents of the ethyl acetate fraction (EtOAcF) obtained from the methanolic extract (ME) was performed by UV-Vis spectrophotometry, infrared spectroscopy, high-performance liquid chromatography (HPLC), mass spectrometry (MS), and cyclic voltammetry, demonstrating the possible majority component of this fraction, the flavone chrysin. Its solubility properties in HPLC are very close to those of the flavonol quercetin, revealing the characteristic presence of this group. An MS spectrum of the fraction revealed a major protonated molecule of m/z 254.9 [M+H]+. The EtOAcF fraction showed three oxidation processes at 0.32 V, 0.54 V, and 0.73 V vs. Ag/AgCl. Three reduction processes at the respective potentials: 0.60 V, −0.03 V, and -0.24 V vs. Ag/AgCl, indicating potential antioxidant activity. At DPPH and ABTS antioxidant radical capture assay, The IC50 obtained was 0.5 mg/mL and 0.81 mg/mL, respectively. In vivo test to determine the mechanical nociceptive threshold in the von Frey test, the dose of 100 mg/kg of the EtOAcF was able to cause inhibition of behavioral changes in neuropathy. The results obtained in this study demonstrate the biological potential of an EtOAcF derived from jaborandi leaves.

The Presence of Some Minor Aspergillus and Penicillium Unregulated Mycotoxins in Main Cereals Cultivated in Albania.

(1) Background: Food and feed safety legislation does not concern all the mycotoxins generated by Penicillium and Aspergillus spp. Certain mycotoxins, including mycophenolic acid (MPA), cyclopiazonic acid (CPA), penicillic acid (PA), roquefortine C (ROQ C), and gliotoxin (GLI), are regarded as having lower toxicity levels, and hence are not included in food and feed legislation. It is obvious that xenobiotics, including mycotoxins, exert synergistic harmful health effects on human and animal when exposed through food and feed. (2) Methods: The presence of these substances in maize and wheat grown in Albania across two consecutive harvesting seasons was investigated by liquid chromatography and mass spectrometry (LC-MS/MS). (3) Results: The findings indicated the presence of these mycotoxins in maize grain but not in wheat grain. In the 2014 season, they exhibited a higher contamination incidence than in the 2015 season. The most commonly detected mycotoxin was MPA, followed by CPA and ROQ C toxin, while PA and GLI were not detected. The MPA revealed a concentration range of 72.9-3447 μg/kg, with a mean value of 1064 μg/kg. Mycophenolic acid was detected in the maize samples collected during the 2015 season. (4) Conclusions: These findings suggest that focusing the investigation only on "controlled" mycotoxins will not produce a proper risk assessment and may not adequately address the possible harmful impacts of mycotoxins on human and animal health due to mycotoxins' co-occurrence.

Using sensory and instrumental analysis to assess the impact of grape smoke exposure on different red wine varietals in California

This study is an investigation of the impact of volatile phenols (VPs) released from burning wood during wildfires on grape composition and the resulting wines. Baseline levels of VPs in grapes and sensory differences between smoke-impacted wines and non-smoke-impacted wines were determined. The differences were related to different levels of smoke taint marker compounds in different wine matrices, using modified descriptive analysis (DA), multivariate statistics, gas chromatography mass spectrometry (GC-MS) and liquid chromatography-triple quadrupole tandem mass spectrometry (LC-QqQ-MS) of the free and total VPs, and individual bound glycosides, respectively. Across two DA panels, Cabernet Sauvignon, Cabernet Franc, Petite Verdot, Merlot, Syrah, Malbec, and Zinfandel wines made from grape originating from different areas in California were evaluated. The results show sensory differences between highly smoke-impacted and non-impacted wines with wines made from highly smoke-impacted grapes characterized as smoky, barbeque, medicinal, and having a retro-nasal ashtray character. Low smoke-impact wines based on free and total VP concentrations were not significantly different from the non-impacted wines when rated through descriptive analysis. The amount of smoke exposure was the largest contributor to smoke impact determined by sensory evaluation, but the different wine matrices from different locations and varietals also played an important role in determining the level of perceived smoke impact. The results of this study will contribute to our understanding of smoke impact and how it influences wine characteristics by relating smoke marker indicator compounds to wine sensory attributes.

Integrated plasma proteomics and myocardial tissue transcriptomics reveal elevated fibrosis markers in arrhythmogenic cardiomyopathy patients

Abstract Background Arrhythmogenic cardiomyopathy (ACM) is characterized by myocardial fibrofatty replacement and ventricular arrhythmias, with potential progression to heart failure. This study aims to identify ACM-specific biomarkers for improved prognosis and possible treatment strategies. Methods RNA sequencing and transcriptome analyses were performed in cardiomyocytes of 10 ACM patients, 10 dilated cardiomyopathy patients and 10 healthy controls. Analyses were performed using liquid chromatography and mass spectrometry. Additionally, plasma protein expression was assessed in 38 ACM patients and 24 healthy controls using the Proximity Extension Assay (Olink®). A standardized bicycle exercise test was performed in 11 ACM patients and controls and blood was obtained before and after exercise. Results Myocardial tissue transcriptomics identified several upregulated molecules associated with extracellular matrix formation and immune response in ACM. Plasma protein sequencing revealed upregulation of proteins involved in metabolic pathways, inflammation, and fibrosis. Integration of these analyses identified key soluble profibrotic markers such as Fibulin-3, Endostatin and C-X-C motif chemokine 10 (CXCL10) in plasma of ACM patients. After exercise, Fibulin-3 levels increased in ACM patients compared to controls. These findings were further validated using enzyme-linked immunosorbent assay (ELISA). Conclusion This comprehensive analysis revealed distinct proteomic and transcriptomic signatures in ACM patients' plasma and cardiomyocytes. The integrative analysis identified several upregulated profibrotic markers in ACM. Our Findings may advance diagnostic and prognostic strategies and offer potential therapeutic targets.

Antioxidant and Anti-Inflammatory Properties of Conceivable Compounds from Glehnia littoralis Leaf Extract on RAW264.7 Cells.

Glehnia littoralis is a medicinal plant, but the scientific basis is still unclear. This study thoroughly investigated phenols from Glehnia littoralis extract (GLE) to determine their potential as anti-inflammatory and antioxidant agents. High-performance liquid chromatography (HPLC) and mass spectrometry (MS) were used to analyze the compounds in GLE. In addition, we performed GLE in vitro in macrophages after lipopolysaccharide (LPS)-induced inflammation. The extract contained eight peaks representing phenolic compounds and one peak representing riboflavin, with the corresponding mass spectrometry data documented. These biologically active compounds were purified by ultrafiltration using LC to determine their ability to target cyclooxygenase-2 (COX-2) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). The results showed that significant compounds were identified, demonstrating a binding affinity for both COX-2 and DPPH. This suggests that the compounds showing excellent binding affinity for COX-2 and DPPH may be the main active ingredients. Vital inflammatory cytokines, including COX-2, inducible nitric oxide synthase (iNOS), mitogen-activated protein kinase (MAPK), and nuclear factor kappa B (NF-κB), were found to be down-regulated during the treatment. In addition, we revealed that the selected drugs exhibited potent binding capacity to inflammatory factors through molecular docking studies. In addition, we confirmed the presence of phenolic components in GLE extract and verified their possible anti-inflammatory and antioxidant properties. This study provided evidence for an efficient strategy to identify critical active ingredients from various medicinal plants. These data may serve as a baseline for further investigations of applying GLE in the pharmaceutical industry.