Using Ultra-high Performance Liquid Chromatography Research Articles

Human metabolism of the semi-synthetic cannabinoids hexahydrocannabinol, hexahydrocannabiphorol and their acetates using hepatocytes and urine samples.

Hexahydrocannabinol (HHC), hexahydrocannabiphorol (HHCP) and their acetates, HHC-O and HHCP-O, respectively, are emerging in Europe as alternatives to tetrahydrocannabinol (THC). This study aimed to elucidate the metabolic pathways of the semi-synthetic cannabinoids HHC, HHCP, HHC-O and HHCP-O from incubation with human hepatocytes. The metabolites of HHC were also identified in authentic urine samples. HHC, HHCP, HHC-O and HHCP-O were incubated with primary human hepatocytes for 1, 3 and 5 h. Authentic urine samples from cases screened positive for cannabis in blood using ELISA but confirmed negative were analysed both non-hydrolysed and hydrolysed for HHC metabolites. Potential metabolites were identified using ultra-high performance liquid chromatography (UHPLC) coupled to a quadrupole time-of-flight mass spectrometer (QToF-MS). HHC and HHCP were primarily metabolised through monohydroxylation (monoOH), followed by oxidation to a carboxylic acid metabolite. HHC-O and HHCP-O were rapidly metabolised to HHC and HHCP, respectively. In authentic urine samples, 18 different metabolites were identified, and 99.3% of hydroxylated metabolites were glucuronidated. 11-OH-HHC, 5'OH-HHC and another metabolite with a monoOH on the side chain were the only metabolites present in all 16 urine samples. The metabolism of HHC and HHCP were similar, although the longer alkyl side chain of HHCP (heptyl) led to greater hydroxylation on the side chain than HHC (pentyl). The use of HHC and HHCP can be differentiated from the use of THC and other phytocannabinoids, but the use of the acetate analogues may not be differentiable from their non-acetate analogues.

Distribution of antibiotic resistance genes and antibiotic residues in drinking water production facilities: Links to bacterial community.

There is a rapid spread of antibiotic resistance in the environment. However, the impact of antibiotic resistance in drinking water is relatively underexplored. Thus, this study aimed to quantify antibiotic resistance genes (ARGs) and antibiotic residues in two drinking water production facilities (NW-E and NW-C) in North West Province, South Africa and link these parameters to bacterial communities. Physicochemical and ARG levels were determined using standard procedures. Residues (antibiotics and fluconazole) and ARGs were quantified using ultra-high performance liquid chromatography (UHPLC) chemical analysis and real-time PCR, respectively. Bacterial community compositions were determined by high-throughput 16S rRNA sequencing. Data were analysed using redundancy analysis and pairwise correlation. Although some physicochemical levels were higher in treated than in raw water, drinking water in NW-E and NW-C was safe for human consumption using the South African Water Quality Guideline (SAWQG). ARGs were detected in raw and treated water. In NW-E, the concentrations of ARGs (sul1, intl1, EBC, FOX, ACC and DHA) were higher in treated water than in raw water. Regarding antimicrobial agents, antibiotic and fluconazole concentrations were higher in raw than in treated water. However, in NW-C, trimethoprim concentrations were higher in raw than in treated water. Redundancy analysis showed that bacterial communities were not significantly correlated (Monte Carlo simulations, p-value >0.05) with environmental factors. However, pairwise correlation showed significant differences (p-value <0.05) for Armatimonas, CL500-29 marine group, Clade III, Dickeya and Zymomonas genera with environmental factors. The presence of ARGs and antibiotic residues in the current study indicated that antibiotic resistance is not only a clinical phenomenon but also in environmental settings, particularly in drinking water niches. Consumption of NW-E and NW-C treated water may facilitate the spread of antibiotic resistance among consumers. Thus, regulating and monitoring ARGs and antibiotic residues in drinking water production facilities should be regarded as paramount.

Study on pulp metabolism of patients with pulpitis using ultra-performance liquid chromatography coupled with Orbitrap mass spectrometry

Background and aimsPulpitis, a pulp disease caused by caries, trauma, and other factors, has a high clinical incidence. This study focused on identifying possible metabolic biomarkers of pulpitis cases and analyzing the related metabolic pathways for providing a theoretical foundation to diagnose and prevent pulpitis. Materials and methodsPulp samples from 20 pulpitis cases together with 20 normal participants were analyzed with a serum metabolomics approach using ultra-high-performance liquid chromatography (UPLC)/Orbitrap mass spectrometry. Moreover, this work carried out multivariate statistical analysis for screening potential biomarkers of pulpitis. ResultsThrough biomarker analysis and identification, such as partial least squares discrimination analysis, orthogonal partial least squares discriminant analysis model establishment, correlation analysis, and biomarker pathway analysis, 40 biomarkers associated with 20 metabolic pathways were identified, including 20 upregulated and 20 downregulated metabolites. Those major biomarkers included oxoglutaric acid, inosine, citric acid, and PA(14:1(9Z)/PGD1). Among them, oxoglutaric acid and inosine were most significantly downregulated and had the highest correlation with pulpitis. Among these metabolic pathways, GABAergic synapse and alanine, aspartate, and glutamate metabolism were positively correlated with pulpitis. 4. ConclusionsThese biomarkers as well as metabolic pathways may offer the theoretical foundation to understand pulpitis pathogenesis and develop preventive drugs.

Non-target analysis of crude oil photooxidation products at high latitudes and their biological effects

With new oil and gas lease sales in high-latitude regions, there exists a need to better understand the chemical fate of spilled oil and its effects on biological life. To address this need, laboratory simulations of crude oil spills under sub-Arctic conditions were conducted using artificial seawater and exposure to solar irradiation to create Hydrocarbon Oxidation Products (HOPs). HOPs characterization and their biological effects were assessed using ultra high-performance liquid chromatography (UHPLC) with high resolution mass Orbitrap spectrometry and the aryl hydrocarbon receptor (AhR) chemically activated luciferase gene expression (CALUX) assay. Non-target UHPLC-Orbitrap mass spectrometry analysis identified 251 HOPs that were in greater abundance in light-exposed samples than dark controls. Oxidized polycyclic aromatic hydrocarbons were also detected, including phenanthrene quinone, anthraquinone, hydroxyanthraquinone, and 9-fluoreneone. The composition of HOPs were consistent with photo-products of alkylated two to four ring PAHs, primarily compounds between 1 and 3 aromatic rings and 1–3 oxygens. The HOP mixture formed during photochemical weathering of Cook Inlet crude oil induced greater AhR activity than parent petroleum products solubilized in dark controls, indicating that HOPs, as a complex mixture, may contribute to petroleum toxicity more than the parent petroleum compounds. These non-targeted approaches provide the most comprehensive analysis of hydrocarbon oxidation products to date, highlighting the diversity of the complex mixture resulting from the photooxidation of crude oil and the limitations of targeted analyses for adequately monitoring HOPs in the environment. Taken together, these data identify a critical “blind spot” in environmental monitoring and spill clean-up strategies as there is a diverse pool of HOPs that may negatively impact human and ecosystem health.

Untargeted metabolomic profiling of 100% malt beers versus those containing barley adjunct

Why was the work done: The incorporation of unmalted barley at high proportion in the grist can introduce unfavourable bitter and astringent characteristics to beer, resulting in an upper limit on the proportion used. The flavour active compounds from raw barley which contribute these characteristics to the beer remain to be identified. How was the work done: This study used non-targeted metabolomics to determine non-volatile metabolites which could contribute to flavour differences when brewing with barley. Three beers were analysed using ultra-high performance liquid chromatography (UHPLC) coupled to a quadrupole time-of-flight mass spectrometer (qTOF) with an electro-spray ionisation source (ESI). One beer was produced using 100% malt and two beers with a grist of 15% barley and 85% malt (beers A and B). The barley was used untreated (beer A) or treated using a proprietary process (beer B). The metabolomic profiles of the three beers were compared and statistically different molecular features were annotated via analysis of MS2 spectra. What are the main findings: Several of the main differential molecular features were nitrogenous peptides and purine derivatives. This was attributed to the lack of the malting process and associated proteolytic enzyme activity reducing the extent of protein and peptide breakdown in the unmalted barley. Several of the identified peptides had amino acid residues which are known to cause bitter and kokumi (rich) taste in beer, which could explain the bitterness when brewing with unmalted barley. Why is the work important: A non-targeted approach offers new insights into non-volatile molecular features in beer that have not been previously identified with targeted analyses. Accordingly, this work identifies metabolites and groups of compounds which have not been previously considered when investigating the unfavourable bitterness and astringency associated with the use of unmalted barley

Electrochemical determination of salbutamol in aqueous solutions with UiO‐66/rGO‐modified electrodes

AbstractThe synthesis of the UiO‐66/rGO material involved combining UiO‐66 with reduced graphene oxide (rGO). This resulting composite found application in the electrochemical determination of salbutamol (SAL) through modification of a glassy‐carbon electrode (GCE) with UiO‐66/rGO. Differential pulse voltammetry served as the chosen technique, offering a wide linear range spanning 1–160 µm and demonstrating a low limit of detection at 0.84 µm. To ensure the reliability of the electrode, various aspects such as sensitivity, repeatability, reproducibility, and stability were meticulously studied. When applied to the analysis of SAL in swine urine solutions (n = 3), the electrode exhibited good recovery rates. Remarkably, the results obtained through this method correlated well with those acquired using ultra‐high performance liquid chromatography (UPLC).

Determination of chiral prothioconazole and its chiral metabolite in water, juice, tea, and vinegar using emulsive liquid–liquid microextraction combined with ultra-high performance liquid chromatography

Emulsive liquid–liquid microextraction (ELLME), a simple, rapid, and environmentally friendly technique, was established to identify chiral prothioconazole and its chiral metabolite in water, juice, tea, and vinegar using ultra-high-performance liquid chromatography (UPLC). Environmentally friendly extractant was mixed with pure water to prepare a high-concentration emulsion, which was added to samples to complete the emulsification and extraction in 1 s. Afterward, an electrolyte solution was added to complete the demulsification without centrifugation. ELLME did not use dispersants compared to the familiar dispersive liquid–liquid microextraction (DLLME), thus reducing the use of toxic solvents and avoiding the effect of dispersants on the partition coefficient. The linear range was from 0.01 to 1 mg/L. The limit of detection was 0.003 mg/L. The extraction recoveries ranged from 82.4 % to 101.6 %, with relative standard deviations of 0.7–5.2 %. The ELLME method developed has the potential to serve as an alternative to DLLME.

Metabolomic Analysis and in Vitro Investigation of the Biological Properties of a By-Product Derived from Vicia faba.

By-products from plant sources are recently regarded as a valuable source of bioactive compounds. In this regard, the present study aims to assess the bioactivities of the 70 % MeOH extract obtained from Vicia faba peels and analyze its metabolomic profile. Acetylcholinesterase and carbohydrate metabolizing enzymes inhibitory activities of the plant extract were assayed using quantitative colorimetric tests. Antioxidant activity was estimated by DPPH assay, and cytotoxic activity was evaluated against normal fibroblast skin cells (1-BJ1). Ninety-one metabolites were tentatively identified using ultra-high-performance liquid chromatography (UHPLC) hyphenated with quadrupole-time-of-flight tandem mass spectrometry (QTOF-MS). Most of these compounds were described for the first time in the plant. In addition, catechin, rutin, quercitrin, and rhamnetin were isolated from the plant extract. The plant extract and the isolated compounds possessed no cytotoxic activity on (1-BJ1), while they exhibited anticholinesterase with the highest activity for 70 % MeOH extract (IC50 =120.11 mg/L), antioxidant potential with the highest activity for rutin (90.54±0.73 %), and carbohydrate metabolizing inhibitory activities with the highest activity for rutin. These discoveries imply that V. faba peels might serve as an efficient antioxidant, exhibit anticholinesterase properties, and have the potential for use in managing diabetes, all while avoiding cytotoxicity in normal cells.

Metabolomics reveals that sulfotransferase 1 may regulate colchicine-induced liver injury

Colchicine is widely used to treat gouty arthritis for years. Previous studies showed that colchicine overdose can cause liver damage, yet the mechanism underlying its hepatotoxicity remains unclear. In this study, hepatotoxicity of colchicine was investigated in vivo. Metabolomic analysis of colchicine metabolites and endogenous metabolites was performed using Ultra High Performance Liquid Chromatography (UHPLC) – mass spectrometry (MS). Seventeen metabolites of colchicine were identified, including 3 novel sulfated metabolites. Meanwhile, endogenous sulfated metabolites were found to be decreased by colchicine. Colchicine might regulate sulfotransferase 1 (SULT1) through perixisome proliferation-activated receptor ɑ (PPARα), and inhibition of SULT1 reduced the levels of sulfated metabolites of colchicine. Inhibition of SULT1 aggravated colchicine-induced liver injury, whereas activation of SULT1 attenuated its liver injury. The supplementation of endogenous sulfated metabolites indoxyl sulfate (IS) or p-cresol sulfate (PCS) alleviated colchicine-induced liver injury through modulation of the CASPASE-1-gasdermin D (GSDMD) pathway. These results indicated that colchicine might cause hepatotoxicity through inhibition of SULT1 and decreased production of bioactive sulfated endogenous metabolites IS and PCS. Our results provided evidence for potential therapeutic targets and agents to prevent liver injury caused by colchicine. Targeting the SULT1 enzyme and administration of IS and PCS may be useful in alleviating colchicine hepatotoxicity.

Active ingredients of Erhuang Quzhi Granules for treating non-alcoholic fatty liver disease based on the NF-κB/NLRP3 pathway

Erhuang Quzhi Granules (EQG), the Chinese herbal compound, has demonstrated significant clinical efficacy in treating non-alcoholic fatty liver disease (NAFLD). The mechanism of this treatment has been shown to involve the nuclear factor kappa B (NF-κB)/nod-like receptor thermal protein domain associated protein 3 (NLRP3) pathway. However, research on the material basis of EQG against NAFLD is still in its primary stages. Following these considerations, this study predicted and screened the active ingredients of EQG using the absorption, distribution, metabolism, and excretion (ADME) property evaluation tool and molecular docking. Then the levels of these active ingredients in EQG were measured using ultra-high-performance liquid chromatography (UHPLC). The efficacy of the active ingredients and their mechanisms were validated through both in vivo and in vitro experiments. The results indicate that the collected 12 components have favorable metabolic stability, are safe, and have drug-like properties. Aloe-emodin (AE), rhein (RH), curcumin (CUR), emodin (EM), and chrysophanol (CP) showed better binding affinity with TNF-α and Caspase-1 proteins. UHPLC analysis revealed that EQG contains AE, RH, CUR, EM, and CP. Cellular experiments proved that all these five ingredients reduce the accumulation of lipids and reactive oxygen species. In animal models of NAFLD, AE, and RH significantly improved the pathological symptoms of steatosis and fibrosis and reduced the levels of pro-inflammatory factors via the NF-κB/NLRP3 pathway. The results reveal the active ingredients of EQG for treating NAFLD based on the NF-κB/NLRP3 pathway and lay the foundation for the clinical promotion of EQG.

Untargeted Metabolomic Study of Lung Cancer Patients after Surgery with Curative Intent.

Lung cancer (LC) is a leading cause of mortality, claiming more than 1.8 million deaths per year worldwide. Surgery is one of the most effective treatments when the disease is in its early stages. The study of metabolic alterations after surgical intervention with curative intent could be used to assess the response to treatment or the detection of cancer recurrence. In this study, we have evaluated the metabolomic profile of serum samples (n = 110) from preoperative (PRE) and postoperative (POST) LC patients collected at two different time points (1 month, A; 3-6 months, B) with respect to healthy people. An untargeted metabolomic platform based on reversed phase (RP) and hydrophilic interaction chromatography (HILIC), using ultra-high performance liquid chromatography (UHPLC) and mass spectrometry (MS), was applied (MassIVE ID MSV000092213). Twenty-two altered metabolites were annotated by comparing all the different studied groups. DG(14,0/22:1), stearamide, proline, and E,e-carotene-3,3'-dione were found altered in PRE, and their levels returned to those of a baseline control group 3-6 months after surgery. Furthermore, 3-galactosyllactose levels remained altered after intervention in some patients. This study provides unique insights into the metabolic profiles of LC patients after surgery at two different time points by combining complementary analytical methods.

Antidiabetic effect of Ardisia elliptica extract and its mechanisms of action in STZ-NA-induced diabetic rat model via 1H-NMR-based metabolomics

Ethnopharmacological relevanceArdisia elliptica Thunb. (AE) (Primulaceae) is a medicinal plant found in the Malay Peninsula and has been traditionally used to treat diabetes. However, limited studies to date in providing scientific evidence to support the antidiabetic efficacy of this plant by in-vitro and in-vivo models. Aim of the studyTo investigate the anti-hyperglycemic potential of AE through in-vitro enzymatic activities and streptozotocin-nicotinamide (STZ-NA) induced diabetic rat models using proton-nuclear magnetic resonance (1H-NMR)-based metabolomics approach. Materials and methodsAnti-α-amylase and anti-α-glucosidase activities of the hydroethanolic extracts of AE were evaluated. The absolute quantification of bioactive constituents, using ultra-high performance liquid chromatography (UHPLC) was performed for the most active extract. Three different dosage levels of the AE extract were orally administered for 4 weeks consecutively in STZ-NA induced diabetic rats. Physical assessments, biochemical analysis, and an untargeted 1H-NMR-based metabolomics analysis of the urine and serum were carried out on the animal model. ResultsType 2 diabetes mellitus (T2DM) rat model was successfully developed based on the clear separation observed between the STZ-NA induced diabetic and normal non-diabetic groups. Discriminating biomarkers included glucose, citrate, succinate, allantoin, hippurate, 2-oxoglutarate, and 3-hydroxybutyrate, as determined through an orthogonal partial least squares-discriminant analysis (OPLS-DA) model. A treatment dosage of 250 mg/kg body weight (BW) of standardized 70% ethanolic AE extract mitigated increase in serum glucose, creatinine, and urea levels, providing treatment levels comparable to that obtained using metformin, with flavonoids primarily contribute to the anti-hyperglycemic activities. Urinary metabolomics disclosed that the following disturbed metabolism pathways: the citrate cycle (TCA cycle), butanoate metabolism, glycolysis and gluconeogenesis, pyruvate metabolism, and synthesis and degradation of ketone bodies, were ameliorated after treatment with the standardized AE extract. ConclusionsThis study demonstrated the first attempt at revealing the therapeutic effect of oral treatment with 250 mg/kg BW of standardized AE extract on chemically induced T2DM rats. The present study provides scientific evidence supporting the ethnomedicinal use of Ardisia elliptica and further advances the understanding of the fundamental molecular mechanisms affected by this herbal antidote.

The analysis of the migration of per and poly fluoroalkyl substances (PFAS) from food contact materials using ultrahigh performance liquid chromatography coupled to ion-mobility quadrupole time-of-flight mass spectrometry (UPLC- IMS-QTOF)

Per-poly fluoroalkyl substances (PFASs) are a group of synthetic fluorine compounds used in food packaging materials to repel water and fats. This study assessed the chemical migration of PFAS from different food contact materials, including cardboard, recycled cardboard, biopolymer, paper and Teflon trays, from various markets. Migration assays were conducted using Tenax® as a food simulant, which was optimized by subjecting it to three consecutive extractions with 3 mL of ethanol within an hour. The resulting extractions were combined and concentrated to 0.5 mL using a nitrogen stream. The analysis was performed using ultrahigh performance liquid chromatography (UPLC) coupled with ion-mobility (IMS) quadrupole-time-of-flight (QTOF) mass spectrometry, which provided a powerful and novel tool for identifying a library of targets containing collision cross section values (CCS) and increasing confidence in subsequent identifications. Eleven PFAS compounds belonging to the family of perfluorocarboxylic acid, perfluorosulfonic acid and perfluorooctanesulfonamidoacetic acid substances (PFCAs, PFSAs and FOSAAs) were found in packaging samples obtained from China, with migrant concentrations ranging 3.2 and 22.3 μg/kg. In contrast, no detectable levels of PFAS were observed in packaging samples obtained in Spain. All trays tested were deemed to be suitable for use as food contact materials due to the fact that their migrant values were lower than 0.025 mg/kg for PFOA and its salts, and lower than a maximum concentration of 1 mg/kg for PFOA-related compounds.

Identification and quantitative analysis of the chemical constituents of Gandouling tablets using ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry.

Gandouling tablets are used in a clinical agent for the treatment of hepatocellular degeneration; however, their chemical constituents have not been elucidated. Here, we screened and identified the chemical constituents of Gandouling tablets using ultra-high-performance liquid chromatography (UHPLC)-quadrupole time of flight/mass spectrometry. A method for the quality evaluation of Gandouling tablets was developed by combining the UHPLC fingerprints and the simultaneous quantitative analysis of multiple active ingredients. For fingerprint analysis, 20 shared peaks were identified to assess the similarities among the 10 batches of Gandouling tablets and the similarity was>0.9. The levels of nine representative active ingredients were simultaneously determined to ensure consistency in quality. A total of 99 chemical components were identified, including 18 alkaloids, 20 anthraquinones, 13 flavonoids, 11 phenolic acids, 9 polyphenols, 7 phenanthrenes, 5 sesquiterpenes, 3 curcuminoids, 2 lignans, 2 isoflavones, 2 dianthranones, and 7 other components. The retention times, molecular formulae, and secondary fragmentation information of these compounds were analyzed, and the cleavage pathways and characteristic fragments of some of the representative compounds were elucidated. This systematic analysis used to identify the chemical components of Gandouling tablets lays the foundation for its further quality control and research on their pharmacodynamic substances.

An intercomparison study of four different techniques for measuring the chemical composition of nanoparticles

Abstract. Currently, the complete chemical characterization of nanoparticles (&lt; 100 nm) represents an analytical challenge, since these particles are abundant in number but have negligible mass. Several methods for particle-phase characterization have been recently developed to better detect and infer more accurately the sources and fates of sub-100 nm particles, but a detailed comparison of different approaches is missing. Here we report on the chemical composition of secondary organic aerosol (SOA) nanoparticles from experimental studies of α-pinene ozonolysis at −50, −30, and −10 ∘C and intercompare the results measured by different techniques. The experiments were performed at the Cosmics Leaving OUtdoor Droplets (CLOUD) chamber at the European Organization for Nuclear Research (CERN). The chemical composition was measured simultaneously by four different techniques: (1) thermal desorption–differential mobility analyzer (TD–DMA) coupled to a NO3- chemical ionization–atmospheric-pressure-interface–time-of-flight (CI–APi–TOF) mass spectrometer, (2) filter inlet for gases and aerosols (FIGAERO) coupled to an I− high-resolution time-of-flight chemical ionization mass spectrometer (HRToF-CIMS), (3) extractive electrospray Na+ ionization time-of-flight mass spectrometer (EESI-TOF), and (4) offline analysis of filters (FILTER) using ultra-high-performance liquid chromatography (UHPLC) and heated electrospray ionization (HESI) coupled to an Orbitrap high-resolution mass spectrometer (HRMS). Intercomparison was performed by contrasting the observed chemical composition as a function of oxidation state and carbon number, by estimating the volatility and comparing the fraction of volatility classes, and by comparing the thermal desorption behavior (for the thermal desorption techniques: TD–DMA and FIGAERO) and performing positive matrix factorization (PMF) analysis for the thermograms. We found that the methods generally agree on the most important compounds that are found in the nanoparticles. However, they do see different parts of the organic spectrum. We suggest potential explanations for these differences: thermal decomposition, aging, sampling artifacts, etc. We applied PMF analysis and found insights of thermal decomposition in the TD–DMA and the FIGAERO.

Antioxidant activity and inhibitory potential of Hibiscus rosa-sinensis flower extract against the key enzymes relevant for hyperglycemia: in-vitro and in-silico studies

BACKGROUND: The alteration of endogenous antioxidants and generation of reactive oxygen species induce hyperglycemia which may lead to the emergence of chronic problems in diabetes. The key enzymes responsible for hyperglycemia are α-amylase and α-glucosidase. Owing to the various negative consequences caused by synthetic medicines, there is a requirement for natural antioxidants and inhibitors for α-amylase, and α-glucosidase enzymes to attenuate hyperglycemia. This article entails the antioxidant potential as well as the inhibitory potential of the Hibiscus rosa-sinensis flower extracts in contrast to the enzymes involved in regulating hyperglycemia.METHODS: Flower extracts were prepared using solvents with ascending polarity viz. hexane, chloroform, ethyl acetate, n-butanol, methanol, and distilled water by maceration. Antioxidant capacity was investigated using 2,2-diphenyl-1-picrylhydrazyl free radical activity, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid cation(s) scavenging assay, ferric ion reducing-antioxidant power assay, and total antioxidant capacity. The inhibitory efficacy against α-amylase and α-glucosidase enzyme were investigated by performing α-amylase, and α-glucosidase inhibitory assays. To validate the observed results in in-vitro assays, the existence of polyphenolic compounds in the most potent extract was detected using ultra-high-performance liquid chromatography (UHPLC). Furthermore, in-silico analysis of the detected polyphenols in the most potent extract was done to understand the binding energies and docking at different sites of α-glucosidase and α-amylase enzymes, and it was contrasted to standard (acarbose).RESULTS: The most significant amount of antioxidant activity was noticed in the ethyl acetate extract in all the assays. Moreover, the ethyl acetate extract had a stronger suppressant effect on α-glucosidase and α-ainilase enzymes than other extracts and it was more in the case of α-glucosidase (IC50 value 53.33 µg/mL) as compared to α-amylase (IC50 value 83.02 µg/mL) enzymes There was a finding that among the polyphenolic compounds detected from the ethyl acetate extract via UHPLC, in which the docking studies were carried out, caffeic acid showed maximum binding energy with the α-glucosidase enzyme while epicatechin showed maximum binding energy with the α-amylase enzyme.CONCLUSIONS: The whole study concluded that caffeic acid and epicatechin have shown good interactions with concerned enzymes, and hence might contribute to the inhibition of hyperglycemic conditions and other diabetic problems caused by oxidative stress.

Not-So-Sweet Dreams: Plasma and IgG N-Glycome in the Severe Form of the Obstructive Sleep Apnea.

Obstructive sleep apnea (OSA) is a prevalent disease associated with increased risk for cardiovascular and metabolic diseases and shortened lifespan. The aim of this study was to explore the possibility of using N-glycome as a biomarker for the severe form of OSA. Seventy subjects who underwent a whole-night polysomnography/polygraphy and had apnea-hypopnea index (AHI) over 30 were compared to 23 controls (AHI under 5). Plasma samples were used to extract 39 glycan peaks using ultra-high-performance liquid chromatography (UPLC) and 27 IgG peaks using capillary gel electrophoresis (CGE). We also measured glycan age, a molecular proxy for biological aging. Three plasma and one IgG peaks were significant in a multivariate model controlling for the effects of age, sex, and body mass index. These included decreased GP24 (disialylated triantennary glycans as major structure) and GP28 (trigalactosylated, triantennary, disialylated, and trisialylated glycans), and increased GP32 (trisialylated triantennary glycan). Only one IgG glycan peak was significantly increased (P26), which contains biantennary digalactosylated glycans with core fucose. Patients with severe OSA exhibited accelerated biological aging, with a median of 6.9 years more than their chronological age (p < 0.001). Plasma N-glycome can be used as a biomarker for severe OSA.

In Vitro Antimicrobial and Antibiofilm Properties and Bioaccessibility after Oral Digestion of Chemically Characterized Extracts Obtained from Cistus × incanus L., Scutellaria lateriflora L., and Their Combination.

Periodontal diseases are oral inflammatory diseases ranging from gingivitis to chronic periodontitis. Porphyromonas gingivalis is one of the major pathogens responsible for severe and chronic periodontitis. Plant extracts with antimicrobial activity could be considered possible alternatives to chlorhexidine, an antiseptic substance used in oral hygiene thatcan cause bacteria resistance. Here, two commercial extracts obtained from Cistus × incanus L. and Scutellaria lateriflora L. were chemically characterized usingUltra-High-Performance Liquid Chromatography (UHPLC) coupled with a Q-Exactive Hybrid Quadrupole Orbitrap Mass Spectrometer. The extracts were studied for their bioaccessibility after simulated in vitro oral digestion, their antimicrobial activity against P. gingivalis, their protective effects against cellular invasion by P. gingivalis, and their antibiofilm activity. The extracts were found to contain very complex mixtures of polyphenols, which were quite stable after in vitro simulated oral digestion and demonstrated mild, dose-dependent inhibitory activity against P. gingivalis growth. This activity increased with the combination of the two extracts. Moreover, the combination of the extracts induced a reduction in P. gingivalis HaCaT invasiveness, and the reduction in biofilm came to around 80%. In conclusion, a combination of C. incanus and S. lateriflora showed promising effects useful in the treatment of gingivitis.

Multidimensional Chromatographic Fingerprinting Combined with Chemometrics for the Identification of Regulated Plants in Suspicious Plant Food Supplements.

The popularity of plant food supplements has seen explosive growth all over the world, making them susceptible to adulteration and fraud. This necessitates a screening approach for the detection of regulated plants in plant food supplements, which are usually composed of complex plant mixtures, thus making the approach not so straightforward. This paper aims to tackle this problem by developing a multidimensional chromatographic fingerprinting method aided by chemometrics. To render more specificity to the chromatogram, a multidimensional fingerprint (absorbance × wavelength × retention time) was considered. This was achieved by selecting several wavelengths through a correlation analysis. The data were recorded using ultra-high-performance liquid chromatography (UHPLC) coupled with diode array detection (DAD). Chemometric modelling was performed by partial least squares-discriminant analysis (PLS-DA) through (a) binary modelling and (b) multiclass modelling. The correct classification rates (ccr%) by cross-validation, modelling, and external test set validation were satisfactory for both approaches, but upon further comparison, binary models were preferred. As a proof of concept, the models were applied to twelve samples for the detection of four regulated plants. Overall, it was revealed that the combination of multidimensional fingerprinting data with chemometrics was feasible for the identification of regulated plants in complex botanical matrices.

Two in vitro anthelmintic assays of four Paraguayan medicinal plants for proof of concept of the role of polyphenols in their biological activities and LC-HRMS analysis

Ethnopharmacological relevanceGastrointestinal nematodes (GIN) control in small ruminants has relied on the systematic use of synthetic anthelmintics (AH), their effectiveness has been progressively decreasing due to the rise and diffusion of anthelmintic resistances. The most prevalent genera affecting small ruminants were Haemonchus spp., and Trichostrongylus spp. The investigation of new anthelmintics in plants is a highly studied option, especially when it is linked to ethnobotanical knowledge and phenolic compounds. The aims of our studyFour medicinal plants mentioned in traditional medicine were selected to evaluate their anthelmintic proprieties at different stages of the life cycle of GIN, namely: Kyllinga odorata Valh., Cassia occidentalis L., Artemisia absinthium L, and Verbena litoralis Kunth and to explore the role of polyphenols in the AH activity. Materials and methodsTo evaluate the anthelmintic activity in this study, two models of GIN species, namely Haemonchus contortus (Hc) and Trichostrongylus colubriformis (Tc) were selected and tested on two in vitro assays: 1) Larval Exsheathment Inhibition Assay (LEIA) and, 2) Egg Hatch Assay (EHA). To explore the role of tannins and polyphenols in AH activity by comparing the effects of LEIA and EHA with or without polyvinylpolypyrrolidone (PVPP) and to characterize the phytochemical composition of the most active plants using ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS). ResultsC. occidentalis exhibited the highest activity on LEIA (EC50 = 250.42–41.80 μg/mL) and A. absinthium on egg hatching processes (EC50 = 121.70–137.34 μg/mL) in both species of GIN. The inhibition in the development of eggs was from 67.70% to 96.36% on H. contortus, and from 78.87% to 99.65% on T. colubriformis. At the maximal dose, Additionally, it was observed that the AH on eggs varies according to the GIN species: on H. contortus the extracts tested blocked the formation of larvae Ovicidal Effect (% higher OE) and on T. colubriformis they blocked the appearance of L1 larvae, Larvae Failing Eclosion (% higher LFE). After PVPP, a reduction in AH activity on LEIA and EHA was noted, especially with C. occidentalis (87.20–67.00% of larvae exsheathment, (p < 0.05) and 40.51–24.96% of egg hatching, (p > 0.05) of both parasite species. Nine putative features were identified using HRMS and MS/MS after addition of PVPP. ConclusionsThe present study demonstrated that C. occidentalis, A. absinthium, and K. odorata, which parts have been traditionally used as medicinal plants are a valuable source of active compounds with anthelmintic activity. The medicinal use of these plants against GIN parasites was proven by in vitro analysis. Therefore exploration of the secondary metabolites of these plant extracts and testing of isolated fractions of active compounds under in vivo experiments are planned and represent a specific challenge for alternative drug research. Regarding the PVPP, in this study we hypotheses about the standard doses it was not able to completely absorb the polyphenols of extracts of K. odorata, C. occidentalis, and A. absinthium, which would lead to more studies to evaluate the role of this product in the absorption of phenolic compounds.