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An appealing strategy for finding novel bioactive molecules in Nature consists in exploring underrepresented and -studied microorganisms. Here, we investigated the antimicrobial and tumoral anti-proliferative bioactivities of twenty-three marine and estuarine bacteria of the fascinating phylum Planctomycetota. This was achieved through extraction of compounds produced by the Planctomycetota cultured in oligotrophic medium followed by an antimicrobial screening against ten relevant human pathogens including Gram-positive and Gram-negative bacteria, and fungi. Cytotoxic effects of the extracts were also evaluated against five tumoral cell lines. Moderate to potent activities were obtained against Enterococcus faecalis, methicillin-sensitive and methicillin-resistant Staphylococcus aureus and vancomycin-sensitive and vancomycin-resistant Enterococcus faecium. Anti-fungal effects were observed against Trichophyton rubrum, Candida albicans and Aspergillus fumigatus. The highest cytotoxic effects were observed against human breast, pancreas and melanoma tumoral cell lines. Novipirellula caenicola and Rhodopirellula spp. strains displayed the widest spectrum of bioactivities while Rubinisphaera margarita ICM_H10T affected all Gram-positive bacteria tested. LC-HRMS analysis of the extracts did not reveal the presence of any known bioactive natural product, suggesting that the observed activities are most likely caused by novel molecules, that need identification. In summary, we expanded the scope of planctomycetal species investigated for bioactivities and demonstrated that various strains are promising sources of novel bioactive compounds, which reenforces the potential biotechnological prospects offered by Planctomycetota. [ABSTRACT FROM AUTHOR]

Cousinia birandiana is one of the widely spread species of the Asteraceae family. In this study, the total phenolic and flavonoid contents, the inhibitory effect on α-amylase, α-glucosidase, acetylcholinesterase and butyrylcholinesterase enzymes, the antioxidant capacity (by DPPH, ABTS, FRAP, CUPRAC tests) and the phytochemical content (by LC-HRMS) of the ethanol extract and fractions were studied. As a result, it was shown that ethanol extract has better enzyme inhibitory effects on α-amylase, AChE and BChE enzymes with 300.000, 380.605 and 133.567 µg/mL IC50 values. Besides that, the highest phenolic and flavonoid content was found in the ethyl acetate fraction (735.986 mgGAE/gextract and 952.317 mgCA/gextract, respectively). It is observed that the same fraction has better antioxidant activity in DPPH (IC50 =112.701 mg/mL), ABTS (9.494 µM Trolox/gextract) and CUPRAC (1120.454 mgGAE/gextract) tests and the n-hexane fraction has a higher FRAP (1351.199 mmol Fe2+/gextract) antioxidant effect. According to the phytochemical analyses quinic acid, chlorogenic acid and isoquercitrin were found as major compounds in the ethanol extract and fractions. In addition, the same assays were carried out with these major substances. So, this plant can be viewed as a promising bio-resource for pharmaceuticals and biomedical products. [ABSTRACT FROM AUTHOR]

BACKGROUND: This study aims to obtain systematic understanding of the way by which pesticides are metabolized in plants and the influence of this process on plants' metabolism as this process has a key impact on plant‐based food safety and quality. The research was conducted under field conditions, which enabled to capture metabolic processes taking place in plants grown under multihectare cultivation conditions. RESULTS: Research was conducted on three wheat varieties cultivated under field conditions and treated by commercially available preparations (fungicides, herbicides, insecticides, and growth regulator). Plant tissues with distinctions in roots, green parts, and ears were collected periodically during spring–summer vegetation period, harvested grains were also investigated. Sample extracts were examined by chromatographic techniques coupled with tandem mass spectrometry for: dissipation kinetics study, identification of pesticide metabolites, and fingerprint‐based assessment of metabolic changes. CONCLUSION: Tissue type and wheat varieties influenced pesticide dissipation kinetics and resulting metabolites. Metabolic changes of plants were influenced by type of applied pesticide and its concentration in plants tissues. Despite differences in plant metabolic response to pesticide stress during cultivation, grain metabolomes of all investigated wheat varieties were statistically similar. 4‐[cyclopropyl(hydroxy)methylidene]‐3,5‐dioxocyclo‐hexanecarboxylic acid and trans‐chrysantemic acid – metabolites of crop‐applied trinexapac‐ethyl and lambda‐cyhalothrin, respectively, were identified in cereal grains. These compounds were not considered to be present in cereal grains up to now. The research was conducted under field conditions, enabling the measurement of metabolic processes taking place in plants grown under large‐scale management conditions. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Therapeutic monoclonal antibodies (mAbs) are crucial in modern medicine due to their effectiveness in treating various diseases. However, the structural complexity of mAbs, particularly their glycosylation patterns, presents challenges for quality control and biosimilarity assessment. This study explores the use of upper-hinge middle-up (UHMU)-level ultra-high-performance liquid chromatography–high-resolution mass spectrometry (LC-HRMS) analysis to improve N-glycan profiling of mAbs. Two specific enzymes, known as IgG degradation enzymes (IGDEs), were used to selectively cleave therapeutic mAbs above the hinge region to separate antibody subunits for further Fc glycan analysis by means of the UHMU/LC-HRMS workflow. The complexity of the mass spectra of IGDEs-digested mAbs was significantly reduced compared to the intact MS level, enabling reliable assignment and relative quantitation of paired Fc glycoforms. The results of the UHMU/LC-HRMS analysis of nine approved therapeutics highlight the significance of this approach for in-depth glycoform profiling. [ABSTRACT FROM AUTHOR]

Achalypha indica is a weed that has medicinal properties. This study aims to determine the metabolite profile and evaluate the antioxidant activity of A. indica from areas with different altitudes, Tuban (20 to 50 masl; TBN), Banyuwangi (60 to 90 masl; BB), Magetan (300 to 600 masl; MGT), and Malang (500 to 800 masl; MLG). The ethanol extract of A. indica was analyzed for total phenolics and antioxidant activity. Separation and identification of samples were carried out using liquid chromatography-high-resolution mass spectrometry (LC HRMS), then classified by Principal Component Analysis (PCA). The total phenol value ranged from 6.83 to 10.67 mg GAE /g of dry powder, with an antioxidant IC 50 of 86.09 to 118.96 µg/mL. LC HRMS successfully identified 83 metabolites. PCA analysis was able to clearly distinguish each sample based on the altitude of the growing location with a total variance value reaching 82 % (PC-1: 57 %, and PC-2: 25 %). Heat map analysis that observes variations in metabolite composition in each sample shows an increase in the content of flavonoid compounds in the lowlands under certain geographical conditions. Therefore, the height of the growing location has a significant influence on the metabolite profile and antioxidant activity. These findings are very important for assessing the quality, safety, and efficacy of medicinal plant raw materials. [Display omitted] • Different altitude can affect the metabolite profile and biological activity in Acalypha indica. • Separation and identification of metabolite using LC HRMS. • Putative metabolite identification by MS2 spectra. • Determination of antioxidant using DPPH methode. • Using chemometrics for PCA and heatmap analysis. [ABSTRACT FROM AUTHOR]

Sesquimustard (Q) is a powerful blistering agent that contains additional sulfur atoms. Sulfur mustard causes covalent bonding by alkylating nucleophilic groups of biologically important macromolecules such as lipids, proteins, DNA, or RNA. Most cells maintain relatively high amounts of a unique tripeptide called glutathione (GSH) (γ-glutamyl-cysteinyl glycine), which possesses a free thiol group, to prevent unwanted reactions caused by reactive chemical entities. Moreover, these thiol groups on cysteines (Cys) are the main target for alkylation. Although Q is the most potent vesicant among sulfur mustards, research studies identifying biomarkers of Q are very limited. Therefore, here in this study, we aimed to identify the GSH and Cys conjugates of Q using mass spectrometric methods and to observe the formation of these conjugates in HaCat cell culture following exposure to different doses. We identified four different conjugates of Q, which are bis-glutathionyl ethylthioethylthioethyl conjugate (GSH-ETETE-GSH), hydroxyethylthioethylthioethyl glutathione conjugate (HETETE-GSH), bis-cysteinyl ethylthioethylthioethyl conjugate (Cys-ETETE-Cys), and hydroxyethylthioethylthioethyl cysteine conjugate (HETETE-Cys). The identity of the conjugates was elucidated using liquid chromatography–high-resolution mass spectrometry (LC-HRMS). We also investigated changes in conjugate formation with exposure concentration and time elapsed after exposure in the cell culture. After exposure, GSH conjugates decreased until 1st hour, while Cys conjugates increased until 6th hour. We also observed that conjugate formation depended on the concentration of Q. This is the first study to elucidate the conjugates of Q dependent on GSH conjugation. As biomarkers are essential tools for evaluating exposure to Q, this study contributes to the limited number of studies identifying biomarkers for Q. [ABSTRACT FROM AUTHOR]

Meat authentication is very important to avoid adulteration, substitution, and mislabeling of meats and meat-based products to protect consumers by ensuring quality, safety, and halal status. This research aimed to employ metabolomics approach using liquid chromatography-high resolution mass spectrometry (LC-HRMS) to identify metabolites of beef (BM), chicken meat (CM), and wild boar meat (WBM) as well as to identify the discriminating metabolites of BM-WBM and CM-WBM. The chemometrics of principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) were used to differentiate BM, CM, and WBM. The orthogonal projection to latent structures-discriminant analysis (OPLS-DA) was used to discriminate and identify discriminating metabolites of BM-WBM and CM-WBM through the variable importance for projections (VIP) value analysis (VIP>1.50, p<0.05). The heatmap plot showed the distribution of discriminating metabolites in BM, CM, and WBM samples. The results of this study suggested that untargeted LC-HRMS successfully identified metabolites in meats. In addition, chemometrics could be used to discriminate between BM, CM, and WBM clearly. In summary, the combination of LC-HRMS and chemometrics is promising for the authentication of meats to ensure the quality as well as halal status of meats. [ABSTRACT FROM AUTHOR]

Bank filtration supports the growing global demand for drinking water amidst concerns over organic micropollutants (OMPs). Efforts to investigate, regulate and manage OMPs have intensified due to their documented impacts on ecosystems and human health. Non-targeted analysis (NTA) is critical for addressing the challenge of numerous OMPs. While identification in NTA typically prioritises compounds based on properties like toxicity, considering substance quantity, occurrence frequency and exposure duration is essential for comprehensive risk management. A prioritisation scheme, drawing from intensive sampling and NTA of bank filtrate, is presented and reveals significant variability in OMP occurrence. Quasi-omnipresent substances, though only 7% of compounds, accounted for 44% of cumulative detections. Moderately common substances, constituting 31% of compounds, accounted for 50% of cumulative detections. Rare compounds, comprising 61%, contributed only 6% to cumulative detections. The application of suspect screening for 31 substances to the dataset yielded results akin to NTA, underscoring NTA's value. Correlation between both methods demonstrates the efficacy of high-resolution mass spectrometry-based NTA in assessing temporal and quantitative OMP dynamics. [ABSTRACT FROM AUTHOR]

In this work we examined the contents of 14 polyphenols in white lupin (Lupinus albus L.) samples cultivated in Southern Italy by the optimisation and validation of a LC-HRMS method. The validation of the LC-HRMS method showed linearity results r2 > 0.989 and recovery values between 71 and 119% for a very wide range of concentrations. Ellagic acid was the most abundant polyphenol, with mean concentrations of 16271.86 ± 19798.53 μg/Kg, followed by apigenin (2749.51 ± 889.95 μg/Kg). A significant variability in ellagic acid contents was found between the areas of cultivation examined (p < 0.05). As far as we know, this work provides the first data on the polyphenols contents of white lupins cultivated in Italy. The comparison with other study confirms the role of the cultivation area for the determination of the polyphenol's contents. The study also confirms white lupins as a promising source of antioxidant and anti-inflammatory substances in a balanced diet. [ABSTRACT FROM AUTHOR]

The concept of extractables and leachable has introduced a new era for identifying potential impurities in drug products. Pharmaceutical packaging materials encompass a variety of polymers due to their appealing properties for storing the drug product. However, numerous chemical species may leach into the drug product from these polymers, posing significant health hazards in the public domain. Identifying such leachable is crucial for assessing safety and addressing toxicological concerns. Acrylic acids are commonly used materials for adhering needles to the barrel in pre-filled syringes. In this study, we identified acrylic acid leachable impurities in ganirelix drug products available in the market using mass spectrometry as an analytical technique. These impurities leached into the drug product during storage conditions. Characterization of the impurities was carried out through data obtained from HRMS and MS/MS analysis, revealing them as polyacrylic acid-ganirelix adduct impurities. This study offers valuable insights into identifying leachable and susceptible sites, providing a foundation for potential modifications in similar classes of drug products, thereby enhancing their safety and efficacy. [ABSTRACT FROM AUTHOR]

Abstract Background This study investigated the antidiabetic and antioxidant properties of hemp seed oil using various bioanalytical methods. Furthermore, this study determined the suppressive properties of hemp seed oil on α-amylase, acetylcholinesterase and carbonic anhydrase II that purified by the sepharose-4B-L-Tyrosine-sulfanilamide affinity chromatoghraphy, all of which are related to different metabolic diseases. Moreover, the phenolic concentration in the essential oil was quantified through LC–HRMS chromatography. Thirteen distinct phenolic compounds were detected in hemp seed oil. Additionally, both the chemical components and quantity of essential oils within hemp seed oil were assessed through GC–FID and GC/MS analyses. Results The predominant essential oils in hemp seed oil included linoleoyl chloride (34.62%), linoleic acid (33.21%), and 2-4-di-tert-butylphenol (5.79%). Hemp seed oil's ability to scavenge radicals was studied through the use of 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazil bioanalytical radical scavenging methods. The results unveiled its potent radical-scavenging properties, with an 46.20 μg/mL for 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals and IC50 of 9.76 μg/mL for 1,1-diphenyl-2-picrylhydrazil radicals. The investigation also extended to explore the reducing capabilities of Fe3+-2,4,6-tri(2-pyridyl)-S-triazine, copper (Cu2+), and iron (Fe3+). Hemp seed oil demonstrated notable inhibitory effect against α-amylase (IC50: 545.66 μg/mL), achethylcholinesterase (IC50: 28.00 μg/mL), and carbonic anhydrase II (IC50: 322.62 μg/mL). Conclusions This interdisciplinary research will prove valuable and set the stage for future investigations into the antioxidant characteristics and enzyme inhibition patterns of plants and plants oils that hold medical and industrial significance.

Abstract Background The high virulence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), responsible for coronavirus disease 2019 (COVID-19), has triggered global health and economic concerns. The absence of specific antiviral treatments and the side effects of repurposed drugs present persistent challenges. This study explored a promising antiviral herbal extract against SARS-CoV-2 from selected Thai medicinal plants based on in vitro efficacy and evaluated its antiviral lead compounds by molecular docking. Methods Twenty-two different ethanolic-aqueous crude extracts (CEs) were rapidly screened for their potential activity against porcine epidemic diarrhea virus (PEDV) as a surrogate using a plaque reduction assay. Extracts achieving ≥ 70% anti-PEDV efficacy proceeded to the anti-SARS-CoV-2 activity test using a 50% tissue culture infectious dose method in Vero E6 cells. Molnupiravir and extract-free media served as positive and negative controls, respectively. Potent CEs underwent water/ethyl acetate fractionation to enhance antiviral efficacy, and the fractions were tested for anti-SARS-CoV-2 performance. The fraction with the highest antiviral potency was identified using liquid chromatography–high-resolution mass spectrometry (LC–HRMS). Molecular docking analyses of these compounds against the main protease (Mpro) of SARS-CoV-2 (6LU7) were performed to identify antiviral lead molecules. The top three hits were further evaluated for their conformational stability in the docked complex using molecular dynamics (MD) simulation. Results The water fraction of mulberry (Morus alba Linn.) leaf CE (WF-MLCE) exhibited the most potent anti-SARS-CoV-2 efficacy with low cytotoxicity profile (CC50 of ~ 0.7 mg/mL), achieving 99.92% in pre-entry mode and 99.88% in postinfection treatment mode at 0.25 mg/mL. Flavonoids and conjugates were the predominant compounds identified in WF-MLCE. Molecular docking scores of several flavonoids against SARS-CoV-2 Mpro demonstrated their superior antiviral potency compared to molnupiravir. Remarkably, myricetin-3-O-β-D-galactopyranoside, maragrol B, and quercetin 3-O-robinobioside exhibited binding energies of ~ − 9 kcal/mol. The stability of each ligand–protein complex of these compounds with the Mpro system showed stability during MD simulation. These three molecules were pronounced as antiviral leads of WF-MLCE. Given the low cytotoxicity and high antiviral potency of WF-MLCE, it holds promise as a candidate for future therapeutic development for COVID-19 treatment, especially considering its economic and pharmacological advantages.

Objective The adulteration of raw beef (BMr) with dog meat (DMr) and pork (PMr) becomes a serious problem because it is associated with halal status, quality, and safety of meats. This research aimed to develop an effective authentication method to detect non-halal meats (dog meat and pork) in beef using metabolomics approach. Methods Liquid chromatography-high resolution mass spectrometry (LC-HRMS) using untargeted approach combined with chemometrics was applied for analysis non-halal meats in BMr. Results The untargeted metabolomics approach successfully identified various metabolites in BMr DMr, PMr, and their mixtures. The discrimination and classification between authentic BMr and those adulterated with DMr and PMr were successfully determined using partial least square-discriminant analysis (PLS-DA) with high accuracy. All BMr samples containing non-halal meats could be differentiated from authentic BMr. A number of discriminating metabolites with potential as biomarkers to discriminate BMr in the mixtures with DMr and PMr could be identified from the analysis of variable importance for projection value. Partial least square (PLS) and orthogonal PLS (OPLS) regression using discriminating metabolites showed high accuracy (R2>0.990) and high precision (both RMSEC and RMSEE

Background: The high virulence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), responsible for coronavirus disease 2019 (COVID-19), has triggered global health and economic concerns. The absence of specific antiviral treatments and the side effects of repurposed drugs present persistent challenges. This study explored a promising antiviral herbal extract against SARS-CoV-2 from selected Thai medicinal plants based on in vitro efficacy and evaluated its antiviral lead compounds by molecular docking. Methods: Twenty-two different ethanolic-aqueous crude extracts (CEs) were rapidly screened for their potential activity against porcine epidemic diarrhea virus (PEDV) as a surrogate using a plaque reduction assay. Extracts achieving ≥ 70% anti-PEDV efficacy proceeded to the anti-SARS-CoV-2 activity test using a 50% tissue culture infectious dose method in Vero E6 cells. Molnupiravir and extract-free media served as positive and negative controls, respectively. Potent CEs underwent water/ethyl acetate fractionation to enhance antiviral efficacy, and the fractions were tested for anti-SARS-CoV-2 performance. The fraction with the highest antiviral potency was identified using liquid chromatography–high-resolution mass spectrometry (LC–HRMS). Molecular docking analyses of these compounds against the main protease (Mpro) of SARS-CoV-2 (6LU7) were performed to identify antiviral lead molecules. The top three hits were further evaluated for their conformational stability in the docked complex using molecular dynamics (MD) simulation. Results: The water fraction of mulberry (Morus alba Linn.) leaf CE (WF-MLCE) exhibited the most potent anti-SARS-CoV-2 efficacy with low cytotoxicity profile (CC50 of ~ 0.7 mg/mL), achieving 99.92% in pre-entry mode and 99.88% in postinfection treatment mode at 0.25 mg/mL. Flavonoids and conjugates were the predominant compounds identified in WF-MLCE. Molecular docking scores of several flavonoids against SARS-CoV-2 Mpro demonstrated their superior antiviral potency compared to molnupiravir. Remarkably, myricetin-3-O-β-D-galactopyranoside, maragrol B, and quercetin 3-O-robinobioside exhibited binding energies of ~ − 9 kcal/mol. The stability of each ligand–protein complex of these compounds with the Mpro system showed stability during MD simulation. These three molecules were pronounced as antiviral leads of WF-MLCE. Given the low cytotoxicity and high antiviral potency of WF-MLCE, it holds promise as a candidate for future therapeutic development for COVID-19 treatment, especially considering its economic and pharmacological advantages. [ABSTRACT FROM AUTHOR]

The recent biological invasion of box tree moth Cydalima perspectalis on Buxus trees has a major impact on European boxwood stands through severe defoliation. This can hinder further regrowth and threaten survival of populations. In a mesocosm approach and controlled larval density over a 2‐month period, responses of B. sempervirens essential and specialized metabolites were characterized using metabolomics, combining 1H–NMR and LC–MS/MS approaches. This is the first metabolome depiction of major Buxus responses to boxwood moth invasion. Under severe predation, remaining green leaves accumulate free amino acids (with the noticeable exception of proline). The leaf trans‐4‐hydroxystachydrine and stachydrine reached 10–13% and 2–3% (DW), while root content was lower but also modulated by predation level. Larval predation promoted triterpenoid and (steroidal) alkaloid synthesis and diversification, while flavonoids did not seem to have a relevant role in Buxus resistance. Our results reveal the concomitant responses of central and specialized metabolism, in relation to severity of predation. They also confirm the potential of metabolic profiling using 1H–NMR and LC–MS to detect re‐orchestration of metabolism of native boxwood after severe herbivorous predation by the invasive box‐tree moth, and thus their relevance for plant–insect relationships and ecometabolomics. [ABSTRACT FROM AUTHOR]

Pinacolyl alcohol (PA), a key forensic marker for the nerve agent Soman (GD), is a particularly difficult analyte to detect by various analytical methods. In this work, we have explored the reaction between PA and 1,1′‐carbonyldiimidazole (CDI) to yield pinacolyl 1H‐imidazole‐1‐carboxylate (PIC), a product that can be conveniently detected by gas chromatography–mass spectrometry (GC–MS) and liquid chromatography‐high‐resolution mass spectrometry (LC‐HRMS). Regarding its GC–MS profile, this new carbamate derivative of PA possesses favorable chromatographic features such as a sharp peak and a longer retention time (RT = 16.62 min) relative to PA (broad peak and short retention time, RT = 4.1 min). The derivative can also be detected by LC‐HRMS, providing an avenue for the analysis of this chemical using this technique where PA is virtually undetectable unless present in large concentrations. From a forensic science standpoint, detection of this low molecular weight alcohol signals the past or latent presence of the nerve agent Soman (GD) in a given matrix (i.e., environmental or biological). The efficiency of the protocol was tested separately in the analysis and detection of PA by EI‐GC–MS and LC‐HRMS when present at a 10 μg/mL in a soil matrix featured in the 44th PT and in a glycerol‐rich liquid matrix featured in the 48th Official Organization for the Prohibition of Chemical Weapons (OPCW) Proficiency Test when present at a 5 μg/mL concentration. In both scenarios, PA was successfully transformed into PIC, establishing the protocol as an additional tool for the analysis of this unnatural and unique nerve agent marker by GC–MS and LC‐HRMS. [ABSTRACT FROM AUTHOR]

Carbendazim, a common pesticide, has shown signs of causing cancer, infertility and toxicity to organisms. Due to its intense use, this fungicide has become a persistent compound in the environment, raising the importance of better understanding its behaviour, metabolic pathways and effects on organisms. Zebrafish is considered an excellent animal model, being able to rapidly absorb compounds in water, mimicking what occurs in the aquatic environment. Therefore, the aim of this work was to evaluate carbendazim metabolites in zebrafish water tank using liquid chromatography coupled to a high-resolution mass spectrometer (LC-HRMS) to highlight analytical targets in order to monitor carbendazim exposure in aquatic environments. For this purpose, treatment, negative and stability groups were defined. In addition, water samples were collected from the tanks and analyses were carried out by LC-HRMS. The results allowed the putative annotation of 7 target metabolites. This study applied the analysis of zebrafish water tank for evaluation of target metabolites of carbendazim as a promising approach, since it is a much cleaner matrix than the usual biological matrices. These metabolites can ensure detectability and further strengthen confidence in monitoring carbendazim exposure in aquatic environments. To the best of our knowledge, there is no one study that has evaluated carbendazim metabolites produced by zebrafish, neither in the animal's body nor in the water tank. This is the first report on the generation of carbendazim metabolites by zebrafish. [ABSTRACT FROM AUTHOR]

This study presents a comprehensive assessment of the performance of popular pitcher-type point-of-use (POU) water filters to remove PFAS contaminants from tap waters. The evaluated filters, Brita (Elite and Standard), Zero Water, Aquagear, and Clearly Filtered, were tested for their efficacy in removing 75 targeted PFAS, total organic carbon (TOC), total dissolved solids (TDS), chloride, and sulfate from two Canadian tap waters with background Σ75 PFAS concentrations of 13 and 56 ng/L. Overall, the performances of the filters varied depending on the structure of the filter media, the water source, and the specific contaminants present. ZeroWater was the top performer in the case of total PFAS removal. The volume-weighted average removal of total PFAS after 160 L of filtration using Saint-Donat tap water was 99% for Zero Water, 99% for Clearly Filtered, 77% for Aquagear, and 20% for Brita (Elite). In the case of Montreal tap water, which had different water characteristics and lower total PFAS levels, the volume-weighted average removal for PFAS was ≈100% for Zero Water, 96% for Clearly Filtered, 60% for Aquagear, 48% for Brita (Elite), and 38% for Brita (Standard). Both laboratory and home tests involving Zero Water filters yielded similar high-performance results using Montreal tap water. Although Zero Water exhibited high PFAS removal (99%) in Saint-Donat water, TDS and TOC desorption and a significant drop in pH were observed after 80 L, a phenomenon which was explained by the higher total concentration of anions in this water. In contrast, no desorption was observed in Montreal tap water for TDS and TOC due to the lower concentrations of anions. The Aquagear filter demonstrated an unusual increase in concentrations of sulfate after the initial 20 L, which needs further evaluation. This study discusses individual filter performance, the influence of tap water characteristics, and the potential to meet the new NSF guidelines, which provides valuable insights for consumers seeking to choose an appropriate easy-to-use water filtration system to ensure safe and clean drinking water in different regions. [ABSTRACT FROM AUTHOR]

Introduction: Untargeted metabolomics studies are expected to cover a wide range of compound classes with high chemical diversity and complexity. Thus, optimizing (pre-)analytical parameters such as the analytical liquid chromatography (LC) column is crucial and the selection of the column depends primarily on the study purpose. Objectives: The current investigation aimed to compare six different analytical columns. First, by comparing the chromatographic resolution of selected compounds. Second, on the outcome of an untargeted toxicometabolomics study using pooled human liver microsomes (pHLM), rat plasma, and rat urine as matrices. Methods: Separation and analysis were performed using three different reversed-phase (Phenyl-Hexyl, BEH C18, and Gold C18), two hydrophilic interaction chromatography (HILIC) (ammonium-sulfonic acid and sulfobetaine), and one porous graphitic carbon (PGC) columns coupled to high-resolution mass spectrometry (HRMS). Their impact was evaluated based on the column performance and the size of feature count, amongst others. Results: All three reversed-phase columns showed a similar performance, whereas the PGC column was superior to both HILIC columns at least for polar compounds. Comparing the size of feature count across all datasets, most features were detected using the Phenyl-Hexyl or sulfobetaine column. Considering the matrices, most significant features were detected in urine and pHLM after using the sulfobetaine and in plasma after using the ammonium-sulfonic acid column. Conclusion: The results underline that the outcome of this untargeted toxicometabolomic study LC-HRMS metabolomic study was highly influenced by the analytical column, with the Phenyl-Hexyl or sulfobetaine column being the most suitable. However, column selection may also depend on the investigated compounds as well as on the investigated matrix. [ABSTRACT FROM AUTHOR]

The possibility of cyanoacetohydrazide usage as a novel derivatizing agent is demonstrated in the presented article, and a comparison with hydroxylamine as the most commonly used reagent is provided. Optimal conditions for steroid derivatization with cyanoacetohydrazide are provided. According to the collected data, the maximum yield of derivatives was observed at pH 2.8 within 70 min at 40 °C with 5 ng/mL limit of detection for all investigated analytes. It was shown that cyanoacetohydrazide derivatives produces both syn- and anti-forms as well as hydroxylamine, and their ratios were evaluated and shown in presented work. An efficiency enchantment from two to up to five times was achieved with a novel derivatization reagent. Its applicability for qualitative analysis of steroids in urine was presented at real samples. Additionally, the reproducible fragmentation of the derivatizing agent in collision-induced dissociation offers opportunities for simplified non-targeted steroidomic screening. Furthermore, cyanoacetohydrazide increases ionization efficiency in positive mode, which can eliminate the need for redundant high-resolution instrument runs required for both positive and negative mode analyses. [ABSTRACT FROM AUTHOR]

The leaves of Araucaria cunninghamii are known to be nonedible and toxic. Previous studies have identified biflavones in various Araucaria species. This study aimed to investigate the in vitro cytotoxicity of the isolated compounds from Araucaria cunninghamii after metabolomics and network pharmacological analysis. Methanol extract of Araucaria cunninghamii leaves was subjected to bioassay‐guided fractionation. The active fraction was analyzed using LC‐HRMS, through strategic database mining, by comparing the data to the Dictionary of Natural Products to identify 12 biflavones, along with abietic acid, beta‐sitosterol, and phthalate. Eight compounds were screened for network pharmacology study, where in silico ADME analysis, prediction of gene targets, compound‐gene‐pathway network and hierarchical network analysis, protein–protein interaction, KEGG pathway, and Gene Ontology analyses were done, that showed PI3KR1, EGFR, GSK3B, and ABCB1 as the common targets for all the compounds that may act in the gastric cancer pathway. Simultaneously, four biflavones were isolated via chromatography and identified through NMR as dimeric apigenin with varying methoxy substitutions. Cytotoxicity study against the AGS cell line for gastric cancer showed that AC1 biflavone (IC50 90.58 μM) exhibits the highest cytotoxicity and monomeric apigenin (IC50 174.5 μM) the lowest. Besides, the biflavones were docked to the previously identified targets to analyze their binding affinities, and all the ligands were found to bind with energy ≤−7 Kcal/mol. [ABSTRACT FROM AUTHOR]

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Mayotte Island, a French department located in the Mozambique Channel, has for several years been faced with the consumption of "La Chimique" (LC), reputed (but extremely poorly documented) to be a mixture of tobacco and synthetic cannabinoid receptor agonists (SCRAs). One of the objectives of the CHASSE‐MAREE protocol is to assess the composition and heterogeneity of LC products through successive LC sample collection campaigns among users. Currently underway, we present here the first analytical results (samples collected in 2022). Between September and December 2022, 80 samples were collected throughout the island over three periods: 70 in the usual form of LC (small folded papers containing a plant‐like sample, mostly tobacco), 6 powders, and 4 cigarettes. Analysis was performed using liquid chromatography with high‐resolution mass spectrometry. The detected substances (number of detections) included SCRAs (MDMB‐4en‐PINACA [35], ADB‐FUBIATA [25], MDMB‐INACA [16], ADB‐BUTINACA [15], AFUBIATA [11], 4F‐MDMD‐BICA [7], CH‐PIATA [14], 5C‐APINACA [3], BZO‐HEXOXIZID [2], and 4F‐ABINACA [1]), nicotine (68), tetrahydrocannabinol (THC), cannabinol (CBN), and cannabidiol (CBD) (2), medications (amantadine [11], cyamemazine [6], and acetaminophen [3]), and a designer benzodiazepine (bromazolam [4]). The SCRAs currently in use are varied, and the market for "cooks" (those who prepare LC) is dispersed according to where and when samples are collected. These preliminary results will be supplemented by analysis of samples collected in the first half of 2023 and by an improved description of the current panorama of consumption of LC in Mayotte (mapping, effects felt and dependence, etc.). [ABSTRACT FROM AUTHOR]

Oral endocrine therapies (OET) for breast cancer treatment need to be taken over a long period of time and are associated with considerable side effects. Therefore, adherence to OET is an important issue and of high clinical significance for breast cancer patients' caregivers. We hypothesized that a new bioanalytical strategy based on liquid chromatography and high-resolution mass spectrometry might be suitable for unbiased adherence monitoring (AM) of OET. Four different biomatrices (plasma, urine, finger prick blood by volumetric absorptive microsampling (VAMS), oral fluid (OF)) were evaluated regarding their suitability for AM of the OET abemaciclib, anastrozole, exemestane, letrozole, palbociclib, ribociclib, tamoxifen, and endoxifen. An analytical method was developed and validated according to international recommendations. The analytical procedures were successfully validated in all sample matrices for most analytes, even meeting requirements for therapeutic drug monitoring. Chromatographic separation of analytes was achieved in less than 10 min and limits of quantification ranged from 1 to 1000 ng/mL. The analysis of 25 matching patient samples showed that AM of OET is possible using all four matrices with the exception of, e.g., letrozole and exemestane in OF. We were able to show that unbiased bioanalytical AM of OET was possible using different biomatrices with distinct restrictions. Sample collection of VAMS was difficult in most cases due to circulatory restraints and peripheral neuropathy in fingers and OF sampling was hampered by dry mouth syndrome in some cases. Although parent compounds could be detected in most of the urine samples, metabolites should be included when analyzing urine or OF. Plasma is currently the most suitable matrix due to available reference concentrations. [ABSTRACT FROM AUTHOR]

Objective: The adulteration of raw beef (BMr) with dog meat (DMr) and pork (PMr) becomes a serious problem because it is associated with halal status, quality, and safety of meats. This research aimed to develop an effective authentication method to detect non-halal meats (dog meat and pork) in beef using metabolomics approach. Methods: Liquid chromatography-high resolution mass spectrometry (LC-HRMS) using untargeted approach combined with chemometrics was applied for analysis non-halal meats in BMr. Results: The untargeted metabolomics approach successfully identified various metabolites in BMr DMr, PMr, and their mixtures. The discrimination and classification between authentic BMr and those adulterated with DMr and PMr were successfully determined using partial least square-discriminant analysis (PLS-DA) with high accuracy. All BMr samples containing non-halal meats could be differentiated from authentic BMr. A number of discriminating metabolites with potential as biomarkers to discriminate BMr in the mixtures with DMr and PMr could be identified from the analysis of variable importance for projection value. Partial least square (PLS) and orthogonal PLS (OPLS) regression using discriminating metabolites showed high accuracy (R2>0.990) and high precision (both RMSEC and RMSEE <5%) in predicting the concentration of DMr and PMr present in beef indicating that the discriminating metabolites were good predictors. The developed untargeted LC-HRMS metabolomics and chemometrics successfully identified non-halal meats adulteration (DMr and PMr) in beef with high sensitivity up to 0.1% (w/w). Conclusion: A combination of LC-HRMS untargeted metabolomic and chemometrics promises to be an effective analytical technique for halal authenticity testing of meats. This method could be further standardized and proposed as a method for halal authentication of meats. [ABSTRACT FROM AUTHOR]

Evidence of an insulin overdose is very complicated in the medico-legal field. The analysis and subsequent interpretation of results is complex, especially when treating postmortem blood samples. The instability of insulin, the special pre-analytical conditions and the absence of specific analytical methods has led most laboratories not to analyze insulin in their routine with a consequent underestimation of cases. This paper aims to assess the difficulties associated with the analytical characterization of insulin by describing a case that typically represents most of the inconveniences encountered following a suspected insulin overdose. The case concerns a man found dead at home by his brother. After an external examination, which did not reveal a specific cause of death, toxicological analysis was requested which did not reveal any substance of toxicological interest. Only 9 months later, it was reported to the toxicologist that the subject was diabetic, on insulin lispro treatment and that three empty syringes were found next to his body. Following analysis by LC-high-resolution mass spectrometry, the presence of insulin lispro at a concentration of 1.1 ng/mL, a therapeutic concentration, was evidenced. Despite the low concentration found, overdose cannot be excluded and this paper will describe the criteria evaluated to reach this conclusion. This case highlights that the interpretation of a postmortem insulin concentration is very complex and requires the evaluation of various elements including the circumstances of death, the subject's medical history, the interval between death and sampling and the sample storage. [ABSTRACT FROM AUTHOR]

Helichrysum armenium subsp. armenium plant was extracted with the Soxhlet apparatus. The obtained extract was fractionated and, respectively, AHB (petroleum ether-60% ethanol-chloroform), AHC (ethanol-toluene), AHD (ethanol-chloroform), and AHE (ethanol-ethylacetate) extracts were obtained. Then, antimicrobial, antioxidant, and acetylcholinesterase activity studies were performed on these extracts. In this study, with the using the LC-HRMS method from the aerial parts of Helichrysum armenium subsp. armenium, the flavonoids: apigenin, apigenin 7-glucoside, apigenin 7-methylate, luteolin 7-glucoside, kaempferol, quercetin, quercitrin, (+)-trans taxifolin, (-)-epicatechin gallate, sinensetin, rutin, 3'-o-methyl quercetin, hispidulin, chrysin, acacetin, genkwanin, chrysoeriol; the phenolic acids: chlorogenic acid, ascorbic acid, fumaric acid, caffeic acid, salicylic acid, caffeic acid phenethyl ester, emodin, vanilic acid; the alkaloid: caffeine and the saponoside; glycyrrhizic acid were obtained. [ABSTRACT FROM AUTHOR]

As first‐line antimalarials used in the artemisinin combination therapy, artemisinin drugs exert their action inside red blood cells. However, the blood pharmacokinetic characteristics of artemisinin drugs have not been fully revealed owing to their built‐in chemical instability initiated by Fe2+ released from hemoglobin, with limited information on their metabolites. In this study, liquid chromatography tandem high‐resolution mass spectrometric (LC–HRMS) methods were developed for the quantification of two representative artemisinin drugs (artemisinin, ART; dihydroartemisinin, DHA) and their respective metabolite (deoxyartemisinin, D‐ART; dihydroartemisinin glucuronide, DHA‐Glu) in rat blood/plasma. The blood samples were pretreated with the stabilizer (0.4 m potassium dichromate and 3% EDTA‐2Na). The methods displayed excellent specificity, linearity, accuracy and precision for ART (17.7–709.2 nm) and its metabolite D‐ART (18.8–751.9 nm), and the linear range was 40.0–4,000.0 nm for both DHA and DHA‐Glu. The methods were successfully applied to the pharmacokinetic studies of ART and DHA in rats. The blood‐to‐plasma ratio was 0.8–1.5 for ART, 1.0–1.5 for D‐ART, 1.2–2.2 for DHA and 0.9–1.3 for DHA‐Glu, which was time dependent. The results indicated that artemisinin drugs and their metabolites showed a high but different blood‐to‐plasma ratio, which should be considered when optimizating their dosing regimens or evaluating their clinical outcomes. [ABSTRACT FROM AUTHOR]

Dibenzoazecines are a new class of potential neuroleptics with a high potential for the treatment of schizophrenia. Initial stress tests indicated that the lead compound, LE404, decomposes when exposed to oxygen and sunlight. In this follow-up study, the influence of oxidative stress and photosensitivity was investigated in accordance with the ICH guidelines. These studies are of great importance for new APIs, as phototoxic and photoallergic reactions pose a significant risk to patients, especially with long-term medication, as it is not possible to completely avoid exposure to sunlight. Furthermore, the identification and prediction of the chemical lability of LE404 can be used to determine suitable storage conditions to prevent compound degradation. The effects of exposure of LE404 to a light source similar in spectrum and intensity to natural sunlight were investigated according to ICH-Q1B. Furthermore, the influence of oxidizing agents was investigated under exclusion of light. Two degradation products were identified. The extent and rate of degradation were continuously monitored using RP-HPLC–UV. Chromatographic separations were performed with a phenomenex™ Gemini 5 µm C18 (250 × 4.60 mm) column and acetonitrile/KH2PO4 buffer (4 mmol L−1, pH 2.5) as mobile phase at 220 nm. The photodegradation product was isolated using semi-preparative RP-HPLC. The oxidation product was obtained by quantitative conversion of LE404 in hydrogen peroxide and subsequent purification by preparative TLC. The structures of both degradation products were elucidated using HR-MS/MS, 1D- and 2D-NMR as well as FT-IR spectroscopy. The characterization of the degradation products serves as the basis for subsequent investigations into their toxicity. [ABSTRACT FROM AUTHOR]

Two untargeted metabolomics approaches (LC-HRMS and 1H NMR) were combined to classify Amarone wines based on grape withering time and yeast strain. The study employed a multi-omics data integration approach, combining unsupervised data exploration (MCIA) and supervised statistical analysis (sPLS-DA).The results revealed that the multi-omics pseudo-eigenvalue space highlighted a limited correlation between the datasets (RV-score = 16.4%), suggesting the complementarity of the assays. Furthermore, the sPLS-DA models correctly classified wine samples according to both withering time and yeast strains, providing a much broader characterization of wine metabolome with respect to what was obtained from the individual techniques. Significant variations were notably observed in the accumulation of amino acids, monosaccharides, and polyphenolic compounds throughout the withering process, with a lower error rate in sample classification (7.52%).In conclusion, this strategy demonstrated a high capability to integrate large omics datasets and identify key metabolites able to discriminate wine samples based on their characteristics.

Human exposure to pesticides in the general population occurs mainly through food consumption. However, specific dietary habits or food products that contribute to pesticide exposure are often unknown. In this study, we propose a combined screening for polyphenols and pesticide residues by liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) to assess the diet and the associated pesticide exposure. We measured 587 urine samples from women around the 34th week of pregnancy of a prospective mother–child cohort. A non-targeted screening for flavonoid-like compounds related to fruit and vegetable consumption was performed, prioritizing 164 features and identifying a total of 46 features by spectral library search. Based on a subset of markers, k-means clustering was performed, leading to four clusters with presumably similar dietary habits. The clusters were compared against food questionnaire data collected within the period of sample collection. Suspect screening of more than 500 pesticide residues including metabolites was performed, with a total of 40 residues being reported for 27 different pesticides. The detection of pesticide residues was compared across the different clusters of dietary habits. Indications were found that pyrimethanil metabolites might be associated with the consumption of citrus fruits or derivate products. We demonstrate that the method used has the potential to reveal patterns of pesticide intake from specific food commodities.

Meat authentication is very important to avoid adulteration, substitution, and mislabeling of meats and meat-based products to protect consumers by ensuring quality, safety, and halal status. This research aimed to employ metabolomics approach using liquid chromatography-high resolution mass spectrometry (LC-HRMS) to identify metabolites of beef (BM), chicken meat (CM), and wild boar meat (WBM) as well as to identify the discriminating metabolites of BM-WBM and CM-WBM. The chemometrics of principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) were used to differentiate BM, CM, and WBM. The orthogonal projection to latent structures-discriminant analysis (OPLS-DA) was used to discriminate and identify discriminating metabolites of BM-WBM and CM-WBM through the variable importance for projections (VIP) value analysis (VIP>1.50, p

IntroductionThe use of illicitly manufactured synthetic opioids, specifically fentanyl and its analogs, has escalated exponentially in the United States over the last decade. Due to the targeted nature of drug detection methods in clinical laboratories and the ever-evolving list of synthetic opioids of concern, alternative analytical approaches are needed.MethodsUsing the fentanyl analog screening (FAS) kit produced by the Centers for Disease Control and Prevention (CDC), we developed a liquid chromatography-high resolution mass spectrometry (LC-HRMS) synthetic opioid spectral library and data acquisition method using information dependent acquisition of product ion spectra. Chromatographic retention times, limits of detection and matrix effects, in urine and serum, for the synthetic opioids in the FAS kit (n = 150) were established. All urine and serum specimens sent to a clinical toxicology laboratory for comprehensive drug testing in 2019 (n = 856) and 2021 (n = 878) were analyzed with the FAS LC-HRMS library to determine the prevalence of fentanyl analogs and other synthetic opioids, retrospectively (2019) and prospectively (2021).ResultsThe limit of detection (LOD) of each opioid ranged from 1 to 10 ng/mL (median, 2.5 ng/mL) in urine and 0.25-2.5 ng/mL (median, 0.5 ng/mL) in serum. Matrix effects ranged from -79 % to 86 % (median, -37 %) for urine, following dilution and direct analysis, and -80 % to 400 % (median, 0 %) for serum, following protein precipitation. The prevalence of fentanyl/fentanyl analogs in serum samples increased slightly from 2019 to 2021 while it remained the same in urine. There were only 2 samples identified that contained a fentanyl analog without the co-occurrence of fentanyl or fentanyl metabolites. Analysis of the established MS/MS spectral library revealed characteristic fragmentation patterns in most fentanyl analogs, which can be used for structure elucidation and drug identification of future analogs.ConclusionsThe LC-HRMS method was capable of detecting fentanyl analogs in routine samples sent for comprehensive drug testing. The method can be adapted to accommodate testing needs for the evolving opioid epidemic.

Background and Aims: Scorzonera ketzkhowelii Sosn. ex Grossh. (Asteraceae) was reported as a new species in the Flora of Turkey in 2010. However, there have been no prior biological or chemical investigations conducted. Methods: This study aims to investigate the antimicrobial activities of the petroleum ether, dichloromethane, ethyl acetate, and n-butanol fractions derived from S. ketzkhowelii, and determination of secondary metabolites by Liquid Chromatography with High Resolution Mass Spectrometry (LC-HRMS) analysis on the most active antimicrobial fractions. Results: Notably, all of the fractions demonstrated antimicrobial activity against selected microorganisms, with the dichloromethane fraction of the aerial part exhibiting a higher inhibition of microbial growth compared to the other extracts. The findings of our study revealed the presence of several phenolic compounds in the dichloromethane fractions from both the aerial and subaerial parts, including 3,4-dihydroxybenzaldehyde, salicylic acid, dihydrocaffeic acid, caffeic acid, caffeic acid phenethyl ester, chlorogenic acid, quercetin, hyperoside, naringenin, apigenin, hispidulin, hispidulin 7-glucoside, chrycin, emodin, and carnosic acid. Furthermore, dichloromethane fraction of the aerial parts contained additional phenolic compounds such as homogentisic acid, verbascoside, (+)-trans taxifolin, apigenin 7-glucoside, luteolin, orientin, and chrysoeriol. Conclusion: The plant's demonstrated antimicrobial attributes and the diverse range of phenolic compounds it contains present it as a promising subject for continued research and potential applications within the realms of medicine and pharmaceuticals. Further exploration of its bioactivity and potential health benefits may reveal novel avenues for its practical utilization. [ABSTRACT FROM AUTHOR]

Pregnant women must be wary of using traditional medicines due to the possibility of their having oxytoxic effects. Indonesia is rich in plants containing antioxidants. One of these plants is Phyllanthus emblica L. This study aims to determine the phytochemical constituents of Phyllanthus emblica L. fruit nanoherbals by LC-HRMS analysis and their antimutagenic activity and teratogenic effects. The study commenced by producing nanoherbal extracts from P. emblica fruit. The phytochemical composition of these extracts was then analyzed using LC-HRMS. The nanoherbal extracts were also tested for their ability to prevent mutations, as indicated by a reduction in micronuclei observed in mouse femur bone marrow smear preparations. The teratogenicity test involved administering the P. emblica fruit nanoherbal at 100, 500, and 1000 mg/kg BW doses. The data were analyzed using SPSS. The phytochemical constituents of the P. emblica fruit nanoherbal include flavonoids, phenols, vitamins, and alkaloids. The P. emblica fruit nanoherbal exhibits antimutagenic activity, as evidenced by a statistical analysis that indicated a significant decrease in the quantity of micronuclei per 200 PCE compared to the negative control (p < 0.05). The administration of the P. emblica fruit nanoherbal at a dosage of 1000 mg/kg BW resulted in a teratogenic impact during the organogenesis stage, as shown by hemorrhage and anomalies in the sternum. [ABSTRACT FROM AUTHOR]

BACKGROUND: Crataegus orientalis Pall. ex M. Bieb fruit (COPMB) is extensively used as a source of various products in the medicinal‐aromatic field and holds the potential for erosion control, ornamental purposes, food source, and economic benefits for forest villagers from its fruits. This study aims to determine the chemical components and biological activities of extracts prepared from COPMB using different solvents. RESULTS: The present work was designed to define the antioxidant activity [phosphomolybdenum (total antioxidant capacity), 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), cupric ion‐reducing antioxidant capacity (CUPRAC) and metal chelating activity (MCA)], phytochemical screening analysis, enzyme inhibitor (α‐amylase, α‐glucosidase and tyrosinase) potential, and liquid chromatography‐high resolution mass spectrometry (LC‐HRMS) secondary metabolite profiling in different extracts of COPMB. The results of LC‐HRMS revealed that fumaric acid was the main phenolic compound in all extracts. Among the extracts, ethyl acetate extract has the highest phytochemical and antioxidant properties [total phenolic content (TPC): 32.5 mg GAE/g, total flavonoid content (TFC): 12.2 mg QE/g, ABTS: 213.0 mg TE/g; CUPRAC: 126.0 mg TE/g, MCA: 145.0 mg EDTA/g; FRAP: 122.8 mg TE/g; TAC: 2.8 mmol TE/g]. Ethyl acetate and methanol extracts are more effective in α‐amylase (0.27 ± 0.01 mg/mL; 0.12 ± 0.00 mg/mL), α‐glucosidase (0.63 ± 0.02 mg/mL; 0.77 ± 0.02 mg/mL) and tyrosinase (0.03 ± 0.00 mg/mL; 0.03 ± 0.00 mg/mL) enzyme inhibition potentials compared to standard acarbose (0.75 ± 0.02 mg/mL for α‐amylase; 1.11 ± 0.03 mg/mL for α‐glucosidase) and kojic acid (0.04 ± 0.00 mg/mL). CONCLUSION: The findings from this study suggest that COPMB could serve as a valuable source of natural agents for the food and pharmaceutical industry. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Marine fish oils (MFOs) have been known for their nutritional compounds, which benefit human health. MFOs contain various lipids which play an essential role in human nutrition. The objective of this research was to apply untargeted lipidomics analysis using liquid chromatography–high-resolution mass spectrometry (LC–HRMS) for the comprehensive identification of lipid compositions extracted from five different marine fishes, namely red snapper (Lutjanus campechanus (A1)), giant grouper (Epinephelus lanceolatus (A2)), baronang crochet (Siganus canaliculatus (B2)), white snapper (Lates calcarifer (C3)), skipjack tuna (Katsuwonus pelamis (D2)). More than 1000 lipid compounds from both positive and negative ionization modes could be detected in each MFO using LC–HRMS untargeted lipidomics analysis. Most lipids were dominated by triglycerides (TG) and diglycerides (DG) classes. Principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) could classify MFO samples in positive and negative ionization modes. Based on variable importance for projections (VIP) value, 15 differentiating lipid compounds were found to have an essential role as potential biomarkers in positive ionization mode. In contrast, ten differentiating lipid compounds were found in negative ionization mode. It can be concluded that untargeted lipidomics using LC–HRMS could be used as a powerful analytical technique for comprehensively identifying lipid compositions in MFO samples. It can be further used to identify the lipid compositions of other fish oil samples. [ABSTRACT FROM AUTHOR]

FAF1 (FAS-associated factor 1) is involved in the activation of Fas cell surface death receptors and plays a role in apoptosis and necrosis. In patients with Parkinson's disease, FAF1 is overexpressed in dopaminergic neurons in the substantia nigra. KM-819, an FAF1 inhibitor, has shown potential for preventing dopaminergic neuronal cell death, promoting the degradation of α-synuclein and preventing its accumulation. This study aimed to develop and validate a quantitative analytical method for determining KM-819 levels in rat plasma using liquid chromatography–tandem mass spectrometry. This method was then applied to pharmacokinetic (PK) studies in rats. The metabolic stability of KM-819 was assessed in rat, dog, and human hepatocytes. In vitro metabolite identification and metabolic pathways were investigated in rat, dog, and human hepatocytes. The structural analog of KM-819, namely N-[1-(4-bromobenzyl)-3,5-dimethyl-1H-pyrazol-4-yl]-2-(phenylsulfanyl) acetamide, served as the internal standard (IS). Proteins were precipitated from plasma samples using acetonitrile. Analysis was carried out using a reverse-phase C18 column with a mobile phase consisting of 0.1% formic acid in distilled water and 0.1% formic acid in acetonitrile. The analytical method developed for KM-819 exhibited linearity within the concentration range of 0.002–10 μg/mL in rat plasma. The precision and accuracy of the intra- and inter-day measurements were <15% for the lower limit of quantification (LLOQ) and all quality control samples. KM-819 demonstrated stability under various sample storage conditions (6 h at room temperature (25 °C), four weeks at −20 °C, three freeze-thaw cycles, and pretreated samples in the autosampler). The matrix effect and dilution integrity met the criteria set by the Food and Drug Administration and the European Medicines Agency. This sensitive, rapid, and reliable analytical method was successfully applied in pharmacokinetic studies in rats. Pharmacokinetic analysis revealed the dose-independent kinetics of KM-819 at 0.5–5 mg/kg, with a moderate oral bioavailability of ~20% in rats. The metabolic stability of KM-819 was also found to be moderate in rat, dog, and human hepatocytes. Metabolite identification in rat, dog, and human hepatocytes resulted in the discovery of six, six, and eight metabolites, respectively. Glucuronidation and mono-oxidation have been proposed as the major metabolic pathways. Overall, these findings contribute to a better understanding of the pharmacokinetic characteristics of KM-819, thereby aiding future clinical studies. [ABSTRACT FROM AUTHOR]

The morphological plasticity of cyanobacteria and their widespread ecological dominance in a wide range of habitats highlights the need for in-depth taxonomic studies. This work focused on the taxonomical revision of Leptolyngbya (Cyanophyceae) strains deposited in the ESSACC culture collection and their metabolomic characterization. Although the studied ESSACC strains were morphologically identified as Leptolyngbya sp., the 16S rRNA gene and 16S–23S rRNA ITS analysis revealed that two strains (LMECYA 079 and LMECYA 173) belong to Haloleptolyngbya and represent a new taxonomical unit, genetically unique, ecologically plastic and adapted to both freshwater and thermal habitats, here described as Haloleptolyngbya lusitanica sp. nov. To perform a suspect screening of cyanometabolites in these strains, we used a non-targeted liquid chromatography-high resolution mass spectrometry (LC-HRMS) metabolomic approach. Several metabolites were identified in Haloleptolyngbya lusitanica: micropeptin MM978, spumigin 640, oscillatoxin A and anabaenopeptin D. Strains were maintained and grown under the same conditions, revealing the common production of oscillatoxin A by both H. lusitanica strains. Other identified metabolites, however, were strain-specific, such as anabaenoptin D, which was only detected in LMECYA 173. The different cyanometabolite profiles reinforce the notion that cyanobacteria have the ability to adapt to different habitats, which is maintained under long-term culturing conditions. [ABSTRACT FROM AUTHOR]

Broccoli (Brassica oleracea L. var. italica Plenck) is a widely consumed vegetable, very popular due to its various nutritional and bioactive components. Since studies on the lipid components of broccoli have been limited so far, the aim of the present work was the study of free fatty acids (FFAs) present in different broccoli parts, aerial and underground. The direct determination of twenty-four FFAs in broccoli tissues (roots, leaves, and florets) was carried out, using a liquid chromatography–high-resolution mass spectrometry (LC-HRMS) method in a 10 min single run. Linolenic acid was found to be the most abundant FFA in all different broccoli parts in quantities ranging from 0.76 to 1.46 mg/g, followed by palmitic acid (0.17–0.22 mg/g) and linoleic acid (0.06–0.08 mg/g). To extend our knowledge on broccoli's bioactive components, for the first time, the existence of bioactive oxidized fatty acids, namely hydroxy and oxo fatty acids, was explored in broccoli tissues adopting an HRMS-based lipidomics approach. 16- and 2-hydroxypalmitic acids were detected in all parts of broccoli studied, while ricinoleic acid was detected for the first time as a component of broccoli. [ABSTRACT FROM AUTHOR]

The European Union requires environmental monitoring of the antidepressant drug venlafaxine. Advanced oxidation processes provide a remedy against the spread of micropollutants. In this study, the photoinduced and electrochemical decompositions of venlafaxine were investigated in terms of mechanism and efficacy using high-performance liquid chromatography coupled to high-resolution multifragmentation mass spectrometry. Kinetic analysis, structure elucidation, matrix variation, and radical scavenging indicated the dominance of a hydroxyl-mediated indirect mechanism during photodegradation and hydroxyl and direct electrochemical oxidation for electrochemical degradation. Oxidants, sulfate, and chloride ions acted as accelerants, which reduced venlafaxine half-lives from 62 to 25 min. Humic acid decelerated degradation during ultra-violet irradiation up to 50%, but accelerated during electrochemical oxidation up to 56%. In silico quantitative structure activity relationship analysis predicted decreased environmental hazard after advanced oxidation process treatment. In general, photoirradiation proved more efficient due to faster decomposition and slightly less toxic transformation products. Yet, matrix effects would have to be carefully evaluated when potential applications as a fourth purification stage were to be considered. [ABSTRACT FROM AUTHOR]

The present study was conducted to compare leaf extracts of Y. gloriosa, Y. treculeana, Y. filamentosa and Y. aloifolia. Plant extracts were obtained by sequential extraction with 50% and 100% methanol using the pressurized liquid extraction method. General comparative analysis of Yucca extracts was performed by 2 D-NMR. The presence of various biologically active components in the extracts was shown, among which steroid saponins predominate. The use of LC-MS and LC-HRMS methods with electrospray ionization made it possible to conduct a comparative analysis of Yucca steroidal saponins. The 33 steroidal saponins of various structures were found. The presence of three characteristic saponins in all Yucca species was shown: yuccaloeside C, yuccaloeside E and yuccaloeside B. The results obtained can be used to control the quality of plant materials and products. [ABSTRACT FROM AUTHOR]

Uterine cancer is the most prevalent gynecologic malignancy in women worldwide. Endometrial cancer (EC) has an 81% five-year survival rate, depending on disease stage and time of diagnosis. While endometrial cancer is largely treatable when detected early, no established screening techniques are available in clinical practice. As a result, one of the most significant issues in the medical field is the development of novel ways for early cancer identification, which could boost treatment success rates. Liquid chromatography–high-resolution mass spectrometry (LC-HRMS)-based metabolomics was employed to explore the metabolomic markers and pathways unique to this cancer type and link them to the benign endometrial hyperplasia that may progress to cancer in 5% to 25% of patients. The study involved 59 postmenopausal participants, 20 with EC type 1, 20 with benign hyperplasia, and 19 healthy participants. Metabolite distribution changes were analyzed, and 338 of these features were dysregulated and significant. The first two main components, PC1 and PC2, were responsible for 11.5% and 12.2% of the total metabolites, respectively. Compared with the control group (CO), EC samples had 203 differentially expressed metabolites (180 upregulated and 23 downregulated); in hyperplasia (HP), 157 metabolites were dysregulated (127 upregulated and 30 downregulated) compared to the CO group while 21 metabolites exhibited differential regulation (16 upregulated and 5 downregulated) in EC plasma samples compared to the HP group. Hyperplasia samples exhibited similar metabolic changes to those reported in cancer, except for alterations in triglyceride levels, 7a,12 b-dihydroxy-5b-Cholan-24-oic acid, and Hept-2-enedioyl carnitine levels. The metabolites N-heptanoyl glycine and -(Methylthio)-2,3-isopentyl phosphate and formimino glutamic acid can be specific markers for hyperplasia conditions and dimethyl phosphatidyl ethanolamine and 8-isoprostaglandin E2 can be specific markers for EC conditions. Metabolic activities rely on mitochondrial oxidative phosphorylation for energy generation. The changes in metabolites identified in our study indicate that endometrial cancer cells adopt alternative strategies to increase energy production to meet the energy demand, thereby supporting proliferation. [ABSTRACT FROM AUTHOR]

Therapeutic monoclonal antibodies (mAbs) are crucial in modern medicine due to their effectiveness in treating various diseases. However, the structural complexity of mAbs, particularly their glycosylation patterns, presents challenges for quality control and biosimilarity assessment. This study explores the use of upper-hinge middle-up (UHMU)-level ultra-high-performance liquid chromatography–high-resolution mass spectrometry (LC-HRMS) analysis to improve N-glycan profiling of mAbs. Two specific enzymes, known as IgG degradation enzymes (IGDEs), were used to selectively cleave therapeutic mAbs above the hinge region to separate antibody subunits for further Fc glycan analysis by means of the UHMU/LC-HRMS workflow. The complexity of the mass spectra of IGDEs-digested mAbs was significantly reduced compared to the intact MS level, enabling reliable assignment and relative quantitation of paired Fc glycoforms. The results of the UHMU/LC-HRMS analysis of nine approved therapeutics highlight the significance of this approach for in-depth glycoform profiling.

High-resolution mass spectrometry (HRMS) has become an important tool for monitoring chemical residues in food, but the time and effort required to evaluate the large amount of data generated by HRMS can be a limiting factor in the widespread application of this tool. Suspect screening, i.e., searching HRMS data against large compound databases or mass lists, represents a practical compromise between using HRMS data to only look for target compounds and performing full non-target analysis. Several different approaches for suspect screening using HRMS data were tested using data from shrimp and eel spiked with veterinary drugs and pesticides as well as from imported aquaculture samples. Most of the analytes (>70%) in the spiked samples were detected and identified by searching against compound databases. To query larger databases and on-line resources such as mzCloud, it was necessary to use software capable of differential analysis and selective filtering, such as Compound Discoverer. Using selective filtering, the number of compounds detected in fish sample extracts could be reduced from tens of thousands to a few hundred by subtracting method blanks and comparing to matrix blank extracts. This smaller list of potential compounds could be further evaluated and compared to available databases and libraries. Analysis of imported aquaculture samples resulted in detection of unexpected contaminants including the dewormer levamisole, the insecticide buprofezin, and potentially the plant alkaloid ricinine. [ABSTRACT FROM AUTHOR]

Eutylone is a cathinone-derived synthetic amphetamine scheduled by the World Health Organization and European Monitoring Centre for Drugs and Drug Addiction since 2022 due to its growing consumption. We report here an eutylone intoxication involving a 38-year-old man and a 29-year-old woman in a chemsex context. A bag containing a white crystalline powder labelled as a research product was found in their vehicle. Nuclear magnetic resonance and liquid chromatography-high-resolution mass spectrometry (LC-HRMS) analyses identified the powder as eutylone and confirmed purity superior to 99%. LC-HRMS data analysis using molecular networking allowed to propose new eutylone metabolites in blood samples in a graphical manner. We described 16 phase I (e.g. hydroxylated or demethylated) and phase II metabolites (glucuroconjugates and sulfoconjugates). The same metabolites were found both in male and female blood samples. Toxicological analyses measured eutylone concentration in blood samples at 1374 ng/mL and 1536 ng/mL for the man and the woman, respectively. A keto-reduced metabolite (m/z 238.144) was synthesized to permit its quantification at 67 ng/mL and 54 ng/mL in male and female blood samples, respectively. Overall, the identification of these metabolites will increase the knowledge of potential drug consumption markers and allow to implement mass spectrometry databases to better monitor future drug abuse or consumption. [ABSTRACT FROM AUTHOR]

Inspiratory muscle training (IMT) is known to promote physiological benefits and improve physical performance in endurance sports activities. However, the metabolic adaptations promoted by different IMT prescribing strategies remain unclear. In this work, a longitudinal, randomized, double-blind, sham-controlled, parallel trial was performed to investigate the effects of 11 weeks (3 days·week−1) of IMT at different exercise intensities on the serum metabolomics profile and its main regulated metabolic pathways. Twenty-eight healthy male recreational cyclists (30.4 ± 6.5 years) were randomized into three groups: sham (6 cm·H2O of inspiratory pressure, n = 7), moderate-intensity (MI group, 60% maximal inspiratory pressure (MIP), n = 11) and high-intensity (HI group, 85–90% MIP, n = 10). Blood serum samples were collected before and after 11 weeks of IMT and analyzed by 1H NMR and UHPLC-HRMS/MS. Data were analyzed using linear mixed models and metabolite set enrichment analysis. The 1H NMR and UHPLC-HRMS/MS techniques resulted in 46 and 200 compounds, respectively. These results showed that ketone body metabolism, fatty acid biosynthesis, and aminoacyl-tRNA biosynthesis were upregulated after IMT, while alpha linolenic acid and linoleic acid metabolism as well as biosynthesis of unsaturated fatty acids were downregulated. The MI group presented higher MIP, Tryptophan, and Valine levels but decreased 2-Hydroxybutyrate levels when compared to the other two studied groups. These results suggest an increase in the oxidative metabolic processes after IMT at different intensities with additional evidence for the upregulation of essential amino acid metabolism in the MI group accompanied by greater improvement in respiratory muscle strength. [ABSTRACT FROM AUTHOR]

The study aimed to evaluate the drug–drug, drug–polyvinyl alcohol, and drug–process incompatibilities for the manufacturing of metformin/sitagliptin fixed dose combinations. The melt manufacturing process is a solvent-free green process such as melt granulation, melt extrusion, and 3D printing by a fusion deposition method. The pre-formulation study was designed to explore compatibility using analytical techniques such as differential scanning calorimeter, high-performance liquid chromatography, and liquid chromatography coupled with high-resolution mass spectra. The drug substances and their physical mixture were subjected to different thermal cycles at 80 °C, 180 °C, and 250 °C. All the samples obtained from the thermal cycles were analyzed for assay by high-performance liquid chromatography. The identification of impurities was carried out by liquid chromatography coupled with high resolution mass spectrometry. The result of differential calorimetry studies reveals that there was no interaction between metformin and sitagliptin in the presence of polyvinyl alcohol up to 180 °C and a potential possibility for the interaction above 180 °C. However, a significant drop in the assay of all the sitagliptin samples at 180 °C was observed in high-performance liquid chromatography, which was further confirmed by mass spectra. The stability of metformin samples at 180 °C was confirmed by all the analytical techniques. The results obtained by this investigation reveal the possibility of manufacturing metformin formulations using the melt manufacturing process, but not for sitagliptin and its combinations. [ABSTRACT FROM AUTHOR]