Using Ultra-high Performance Liquid Chromatography Research Articles

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 (< 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.

Profiling phenolic compounds in juice and peel of fourteen pomegranate (Punica granatum L.) varieties grown in Florida, USA

Pomegranate (Punica granatum L.) fruit contain several health-promoting compounds, but the profiles of these compounds and their variations in nutraceutical value are still lacking. Thus, the profile of phenolic compounds in juice and peel of fourteen pomegranate varieties grown in Florida were evaluated using ultrahigh performance liquid chromatography (UHPLC). A total of 14 phenolic compounds were detected in juice and peel samples and four phenolic compounds were identified and quantitated in studied varieties. Punicalagin A and B were the most representative phenolic compounds in juice of all varieties; the ‘EG’ genotype had the highest content of punicalagin A and B, 93.3 and 238.7 µg/mL, respectively. In peel, ellagic and gallic acids were the most abundant compound; ‘EG’ variety had the highest content of these phenolic compounds, 690.2 and 42.2 µg/mL, respectively. These results can contribute to retrieval and biodiversity valorization of such germplasm by a full characterization of the germplasm.

Effects of Bee Pollen Derived from Acer mono Maxim. or Phellodendron amurense Rupr. on the Lipid Composition of Royal Jelly Secreted by Honeybees.

Royal jelly is a specific product secreted by honeybees, and has been sought after to maintain health because of its valuable bioactive substances, e.g., lipids and vitamins. The lipids in royal jelly come from the bee pollen consumed by honeybees, and different plant source of bee pollen affects the lipid composition of royal jelly. However, the effect of bee pollen consumption on the lipid composition of royal jelly remains unclear. Herein, we examined the influence of two factors on the lipid composition of royal jelly: first, two plant sources of bee pollen, i.e., Acer mono Maxim. (BP-Am) and Phellodendron amurense Rupr. (BP-Pa); secondly, different feeding times. Lipidomic analyses were conducted on the royal jelly produced by honeybees fed BP-Am or BP-Pa using ultra-high performance liquid chromatography (UPLC)-Q-Exactive Orbitrap mass spectrometry. The results showed that the phospholipid and fatty acid contents differed in royal jelly produced by honeybees fed BP-Am compared to those fed BP-Pa. There were also differences between timepoints, with many lipid compounds decreasing in abundance soon after single-pollen feeding began, slowly increasing over time, then decreasing again after 30 days of single-pollen feeding. The single bee pollen diet destroyed the nutritional balance of bee colonies and affected the development of hypopharyngeal and maxillary glands, resulting in differences in royal jelly quality. This study provides guidance for optimal selection of honeybee feed for the production of high-quality royal jelly.

Integration of proteomic and metabolomic analyses: New insights for mapping informal workers exposed to potentially toxic elements.

Occupational exposure to potentially toxic elements (PTEs) is a concerning reality of informal workers engaged in the jewelry production chain that can lead to adverse health effects. In this study, untargeted proteomic and metabolomic analyses were employed to assess the impact of these exposures on informal workers' exposome in Limeira city, São Paulo state, Brazil. PTE levels (Cr, Mn, Ni, Cu, Zn, As, Cd, Sn, Sb, Hg, and Pb) were determined in blood, proteomic analyses were performed for saliva samples (n = 26), and metabolomic analyses in plasma (n = 145) using ultra-high performance liquid chromatography (UHPLC) coupled with quadrupole-time-of-flight (Q-TOF) mass spectrometry. Blood PTE levels of workers, controls, and their family members were determined by inductively coupled plasma-mass spectrometry (ICP-MS). High concentration levels of Sn and Cu were detected in welders' blood (p < 0.001). Statistical analyses were performed using MetaboAnalyst 4.0. The results showed that 26 proteins were upregulated, and 14 proteins downregulated on the welder group, and thirty of these proteins were also correlated with blood Pb, Cu, Sb, and Sn blood levels in the welder group (p < 0.05). Using gene ontology analysis of these 40 proteins revealed the biological processes related to the upregulated proteins were translational initiation, SRP-dependent co-translational protein targeting to membrane, and viral transcription. A Metabolome-Wide Association Study (MWAS) was performed to search for associations between blood metabolites and exposure groups. A pathway enrichment analysis of significant features from the MWAS was then conducted with Mummichog. A total of 73 metabolomic compounds and 40 proteins up or down-regulated in welders were used to perform a multi-omics analysis, disclosing seven metabolic pathways potentially disturbed by the informal work: valine leucine and isoleucine biosynthesis, valine leucine and isoleucine degradation, arginine and proline metabolism, ABC transporters, central carbon metabolism in cancer, arachidonic acid metabolism and cysteine and methionine metabolism. The majority of the proteins found to be statistically up or downregulated in welders also correlated with at least one blood PTE level, providing insights into the biological responses to PTE exposures in the informal work exposure scenario. These findings shed new light on the effects of occupational activity on workers' exposome, underscoring the harmful effects of PTE.

Application of feature-based molecular networking in the field of algal research with special focus on mycosporine-like amino acids

Marine red algae have been known as an excellent source for natural sunscreens and antioxidants for a long time, which outlines their potential for various medical and cosmeceutical applications. This is due to their synthesis of unique secondary metabolites to shield themselves from high levels of UV-A and -B radiation encountered in their natural habitats. In this study, a comprehensive and contemporary way for the detection, visualization, and dereplication of algal natural products with special focus on mycosporine-like amino acids (MAAs) is shown, employing HR-MS/MS metabolomics. 33 crude algal extracts were explored using ultra-high-performance liquid chromatography (UHPLC) hyphenated to orbitrap high-resolution tandem mass spectroscopy (HRMS2). Acquired raw data, subjected to pretreatment and spectral organization, could subsequently be implemented in the Global Natural Products Social (GNPS) workflow, whereby a feature based molecular network (FBMN) was created and visualized in Cytoscape. This FBMN was matched against an in-house as well as open source library on the GNPS platform and additionally enhanced by chemotaxonomic classification software and spectra of standard MAAs, as well as further information layers covering e.g. physicochemical properties, taxonomy, and fragmentation behavior. Based on the integration of the latest in silico annotation tools (SIRIUS, CANOPUS, MSNovelist) as well as already published fragmentation patterns of MAAs, structures for known compounds could be corroborated as well as those for novel substances proposed. This offers an interesting and state-of-the-art approach towards the identification and classification of known and new MAAs.

Herbal formula Jiawei Xiaochengqi decoction prevents postoperative abdominal adhesion in a rat model through inhibition of CXCL2-CXCR2 pathway

BackgroundPostoperative abdominal adhesion (PAA) is the most common complication after abdominal surgeries, which can lead to intestinal obstruction, chronic abdominal pain or female infertility. Jiawei Xiaochengqi decoction (JWXCQ) is a hospital preparation widely used for PAA treatment in Nanfang Hospital of Southern Medical University for more than twenty years. PurposeThis study aimed to investigate the therapeutic effects and potential mechanism of JWXCQ against PAA and provide beneficial information for its clinical application. MethodsThe main active components of JWXCQ were identified using ultra high performance liquid chromatography (UHPLC) combined with standard substance comparison. The efficacy and underlying mechanism of JWXCQ were evaluated through in vivo experiments with a postsurgical-induced peritoneal adhesion rat model, and in vitro studies with LPS-stimulated Raw 264.7 macrophages and primary fibroblasts. H&E and Masson staining were performed to assess histopathological changes. The levels of cytokines/proteins-associated with inflammation and degradation of extracellular matrix as well as CXCL2-CXCR2 pathway-related proteins were determined by ELISA, qRT-PCR, western blot assays or immunohistochemistry, respectively. Furthermore, siCXCR2 transfection was used to validate the mechanism of action of JWXCQ. ResultsJWXCQ treatment significantly reduced the formation of PAA, inhibited the inflammation and collagen deposition, and facilitated the secretion of MMP9, decreased the levels of IL-1β, IL-6, TIMP1, COL-1, and suppressed the CXCL2-CXCR2 pathway in PAA rats. Furthermore, JWXCQ inhibited its downstream pathways, the JAK2-STAT3 and PI3K-AKT signaling, as indicated by the suppression of the phosphorylation levels of STAT3 and AKT. In vitro cell experiments revealed that JWXCQ reduced IL-1β and IL-6 secretion in Raw 264.7 macrophages and COL-1 in primary fibroblasts. The CXCL2-CXCR2, JAK2-STAT3 and PI3K-AKT pathways were also inhibited after JWXCQ treatment, which were consistent with the in vivo results. More importantly, silence of CXCR2 eliminated the regulatory effects of JWXCQ. ConclusionJWXCQ could effectively prevent the PAA formation by alleviating inflammation and collagen deposition, which was associated with the inhibition of CXCL2-CXCR2 pathway. This study investigated the relevant pharmacological mechanisms of JWXCQ, providing further evidence for the application of JWXCQ in clinical PAA treatment.

Assessment of Xenoestrogens in Jordanian Water System: Activity and Identification

Sex hormone disruptors (xenoestrogens) are a global concern due to their potential toxicity. However, to date, there has been no study to investigate the presence of xenoestrogen pollutants in the Jordanian water system. Samples in triplicates were collected from six locations in Jordan, including dams, surface water, tap or faucet water, and filtered water (drinking water-local company). Xenoestrogens were then extracted and evaluated with a yeast estrogen screen utilizing Saccharomyces cerevisiae. Later, possible pollutants were mined using ultrahigh-performance liquid chromatography (UPLC) coupled with a Bruker impact II Q-TOF-MS. Possible hits were identified using MetaboScape software (4000 compounds), which includes pesticide, pharmaceutical pollutant, veterinary drug, and toxic compound databases and a special library of 75 possible xenoestrogens. The presence of xenoestrogens in vegetable samples collected from two different locations was also investigated. The total estrogen equivalents according to the YES system were 2.9 ± 1.2, 9.5 ± 5, 2.5 ± 1.5, 1.4 ± 0.9 ng/L for King Talal Dam, As-Samra Wastewater Treatment Plant, King Abdullah Canal, and tap water, respectively. In Almujeb Dam and drinking water, the estrogenic activity was below the detection limit. Numbers of identified xenoestrogens were: As-Samra Wastewater Treatment Plant 27 pollutants, King Talal Dam 20 pollutants, Almujeb Dam 10 pollutants, King Abdullah Canal 16 pollutants, Irbid tap water 32 pollutants, Amman tap water 30 pollutants, drinking water 3 pollutants, and vegetables 7 pollutants. However, a large number of compounds remained unknown. Xenoestrogen pollutants were detected in all tested samples, but the total estrogenic capacities were within the acceptable range. The major source of xenoestrogen pollutants was agricultural resources. Risk evaluations for low xenoestrogen activity should be taken into account, and thorough pesticide monitoring systems and regular inspections should also be established.

UPLC–MS/MS-Based Targeted Proteomics Assay to Evaluate the Relationship between CYP3A2 Expression and Enzymatic Activity

Traditional analytical methods such as western blotting and reverse transcription-polymerase chain reaction only allow semiquantitative assessments and cannot accurately quantify the expression of the cytochrome P450 (CYP) enzyme. Therefore, the relationship between excess CYP enzyme and enzymatic activity is not been systematically studied yet. In the present study, we developed a new method using ultra-high-performance liquid chromatography (UPLC) mass spectroscopy (MS)/MS and determined the accurate quantification of CYP3A2 in rat liver. The linear range was 0.1329–8.508 ng/mL. The intra- and interday precisions and accuracies data fulfilled the acceptance criteria of food and drug association guidelines for bio analytical method validation. We compared the quantitative results with CYP3A2 activity obtained by the cocktail assay. The developed method was successfully used for the quantitative analysis of CYP3A2. It provided a good background for determining the differences in drug metabolism caused by different CYP expression. Keywords: UPLC-MS/MS; CYP3A2; Accurate Quantitation; Enzyme Activity

Susceptibility of Human Plasma N-glycome to Low-Calorie and Different Weight-Maintenance Diets.

Aberrant plasma protein glycosylation is associated with a wide range of diseases, including diabetes, cardiovascular, and immunological disorders. To investigate plasma protein glycosylation alterations due to weight loss and successive weight-maintenance diets, 1850 glycomes from participants of the Diogenes study were analyzed using Ultra-High-Performance Liquid Chromatography (UHPLC). The Diogenes study is a large dietary intervention study in which participants were subjected to a low-calorie diet (LCD) followed by one of five different weight-maintenance diets in a period of 6 months. The most notable alterations of the plasma glycome were 8 weeks after the subjects engaged in the LCD; a significant increase in low-branched glycan structures, accompanied by a decrease in high-branched glycan structures. After the LCD period, there was also a significant rise in N-glycan structures with antennary fucose. Interestingly, we did not observe significant changes between different diets, and almost all effects we observed immediately after the LCD period were annulled during the weight-maintenance diets period.