Liquid Chromatography Quadrupole Research Articles

Evaluation of organophosphorus pesticide tyrosine adducts for postmortem change by human serum albumin with liquid chromatography quadrupole Orbitrap mass spectrometry.

Organophosphorus pesticides (OPPs) having a phosphate ester moiety, such as malathion (MA) and methidathion (DMTP), are widely used and have been detected in many fatal cases of accidental exposure or suicide in Japan. In forensic toxicology, the accurate determination of blood OPP concentration is mandatory to prove death by OPP poisoning. However, fatal pesticide concentration in blood at autopsy varies depending on the circumstances surrounding the dead body. In this study, we found that 16 OPPs were degraded by human serum albumin (HSA) in a temperature-dependent fashion. The mechanism underlying MA, DMTP, azinphos-methyl, etrimfos, fenthion (MPP), pirimiphos-methyl, (E)-dimethylvinphos, (Z)-dimethylvinphos, vamidothion, edifenphos (EDDP), fosthiazate, and pyraclofos degradation involves the formation of adducts with tyrosine residues in HSA. The mass spectra obtained by liquid chromatography quadrupole Orbitrap mass spectrometry revealed that phosphate ester amino acid adducts such as Y-adduct1, Y-adduct2, Y-adduct3, Y-adduct4, and Y-adduct5 were formed in HSA solution incubated with OPPs. These results indicate that the 16 OPPs are postmortem changed by HSA. The detection of phosphate ester amino acid adducts such as Y-adduct1, Y-adduct2, Y-adduct3, Y-adduct4, and Y-adduct5, instead of MA, DMTP, azinphos-methyl, etrimfos, MPP, pirimiphos-methyl, (E)-dimethylvinphos, (Z)-dimethylvinphos, vamidothion, EDDP, fosthiazate, and pyraclofos per se, may be used to determine death by these OPPs poisoning.

High-resolution Tandem Mass Spectrometry for Studying Chemical Constituents of Gynura bicolor DC.

The separation and analysis of the desired chemical components are important subjects for the fundamental research of traditional Chinese medicine (TCM). Ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) has gradually become a leading technology for the identification of TCM ingredients. Gynura bicolor DC. (BFH), a perennial stemless herb used for medicine and food in China has medicinal effects such as clearing heat, moistening the lung, relieving cough, dispersing stasis, and relieving swelling. Polyphenols and flavonoids contain numerous isomers, which hinder the identification of the complex compounds in BFH. This paper presents a systematic protocol for studying chemical constituents of BFH based on solvent extraction and integrated data via UPLC-Q-TOF-MS. The method described here includes systematic protocols for sample pretreatment, MS calibration, MS acquisition, data processing, and analysis of results. Sample pretreatment includes collection, cleaning, drying, crushing, and extraction. MS calibration consists of multipoint and single-point correction. Data processing includes data importing, method establishment, analysis processing, and result presentation. Representative results of the typical fragmentation pattern of phenolic acids, esters, and glycosides in Gynura bicolor DC. (BFH) are presented in this paper. In addition, organic solvent selection, extraction, data integration, collision energy selection, and method improvement are discussed in detail. This universal protocol can be widely used to identify complex compounds in TCM.

Profiling of 3-Formyl rifamycin (3-FR) from sintered fixed dose combinations (SDCs) of rifampicin and isoniazid by using RP-HPLC, LC-ESI-QTOF and UPLC/MS/MS

The rapid and severe decomposition of rifampicin in the presence of isoniazid (INH) in a gastric acidic pH environment is a well-reported phenomenon. It is important to note that the significant decomposition problem of RIF has an impact on its bioavailability, which becomes a major issue in fixed dose combinations (FDCs). In this degradation process, RIF will undergo acid hydrolysis and form 3-FR, an acid degradation product of RIF. Further, it will react with INH and form an insoluble iso-nicotinoyl hydrazone (HYD). The formed HYD is probably not stable in the acidic condition, so it will further convert into INH and 3-FR. Consequently, the generation of 3-FR in the acidic stomach environment can be a critical factor contributing to a substantial decline in RIF bioavailability and warrants careful consideration. Hence, it is imperative to explore the presence of 3-FR not only in in vitro dissolution samples but also in in vivo plasma samples. Therefore, the current study focused on the chemical analysis of 3-FR using additively manufactured sintered drug combinations (SDCs) to evaluate its formation, release and qualitative absorption in both in vitro and in vivo settings. High-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry (LC-ESI-QTOF-MS/MS) were employed for analytical investigations. The developed LC/MS-MS method was employed to investigate the presence of 3-FR in the rabbit's plasma following a single oral administration of SDCs. The study revealed the significant presence of 3-FR in both the in vitro and in vivo plasma samples obtained from the SDCs of RIF and INH. This finding indicates the potential of altering the strategy through additive manufacturing of pH-dependent SDCs to address the critical issue of RIF.

Metabolomic and chemometric profiles of Tribulus terrestris L. from three different locations in Mpumalanga province, South Africa

Background: Tribulus terrestris L. is a traditional herb regularly recognised as puncture vine, yellow vine, devils horn, goat head and caltrop, this is often a yearly shaggy herbaceous plant species with stems of up to 2m long, having a place to the family of Zygophyllaceae.Aim: The study explored the therapeutic potential of this herb as it is being utilised for pharmaceutical purposes because of its furostanol saponins, which have a stimulating impact on characteristic testosterone levels.Setting: The study took place in different locations of Mpumalanga Province, Bushbuckridge (24.8398°S, 31.0464°E), Kamagugu (25.4566° S, 31.0034° E) and Nkomatipoort (25.4510° S, 31.9587° E).Methods: Utilising ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS), 50 metabolites were tentatively identified within the leaves of Tribulus terrestris L. from three diverse areas of Mpumalanga Province, South Africa.Results: Metabolomic-chemometric analysis revealed that Parvispinoside B, F-Gitonin and Gitonin scored highest for the discrimination of Tribulus terrestris L. from three locale clusters. Heat maps showed designs and groupings based on the metabolite concentrations.Conclusion: This study provided novel insights in terms of thorough identification of the secondary metabolites and characterization of the leaves of Tribulus terrestris L. in the areas studied.Contribution: To the best of our knowledge, this study presents the first metabolite profile of Tribulus terrestris L. and its compositional differences in the Mpumalanga region, providing chemical-based evidence for its nutritive and/or health benefits

Chemical Profiling of Shen-Wu-Yi-Shen Tablets Using UPLC-Q-TOF-MS/MS and Its Quality Evaluation Based on UPLC-DAD Combined with Multivariate Statistical Analysis.

Shen-Wu-Yi-Shen tablets (SWYST) is a traditional Chinese medicine prescription used for treating chronic kidney disease (CKD). This study aims to characterize the constituents in SWYST and evaluate the quality based on the quantification of multiple bioactive components. SWYST samples were analyzed with ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and a data-processing strategy. As a result, 215 compounds in SWYST were unambiguously identified or tentatively characterized, including 14 potential new compounds. Meanwhile, strategies based on characteristic fragments for rapid identification were summarized, indicating that the qualitative method is accurate and feasible. Notably, the glucose esters of laccaic acid D-type anthraquinone were first found and their fragmentation patterns were described by comparing that of O-glycoside isomers. Besides, based on comparisons of the cleavage ways of mono-acyl glucose with different acyl groups or acylation sites, differences in fragmentation pathways between 1,2-di-O-acyl glucose and 1,6-di-O-acyl glucose were proposed for the first time and verified by reference substances. In addition, a validated UPLC-DAD was established for the determination of 11 major bioactive components related to treatment of CKD (albiflorin, paeoniflorin, 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-d-glucoside (TSG), 1-O-galloyl-2-O-cinnamoyl-β-d-glucose, emodin-8-O-β-d-glucoside, chrysophanol-O-β-d-glucoside, aloe-emodin, rhein, emodin, chrysophanol and physcion). Moreover, TSG and 1-O-galloyl-2-O-cinnamoyl-β-d-glucose were found as the quality markers related to the origins of SWYST based on multivariate statistical analysis. Conclusively, the findings in this work provide a feasible reference for further studies on quality research and mechanisms of action in treating CKD.

Nitrogen-doped carbon enhances Fe0 activation for efficient tetrabromobisphenol a decontamination and its removal mechanism

Biomass derived N-doped carbon-encapsulated nano zero-valent iron (NC@Fe0) was successfully used to activate hydrogen peroxide (H2O2) for efficient oxidation of tetrabromobisphenol A (TBBPA). The as-prepared NC@Fe0 exhibited high H2O2 activation performance, with a TBBPA removal efficiency of 95.9 % and an oxidation rate constant of 0.031 min−1. The quenching experiments and electrochemical tests showed that the overall contributions of nonradical pathways were over 83.0 %, via synergistic production of singlet oxygen (1O2) and an electron transfer process (ETP). Specifically, the N group on NC@Fe0 greatly enhanced 1O2 production, while formation of surface complexes (OVs-H2O2) was responsible for the ETP mechanism. High Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometer (HPLC-Q-TOF MS/MS) and Toxicity Estimation Software Tool (T.E.S.T) clearly demonstrated that products were mainly generated via debromination and β-scission, where the products were less toxic than the parent. During practical wastewater, in both lake water and domestic wastewaters, NC@Fe0/H2O2 removed over 91.0 % of TBBPA. This study highlighted the considerable contribution of nonradicals in contaminant removal and enriched our understandings of a green activator for future environmental remediations.

Investigating the mechanism and efficacy material basis of Xiehuo Xiaoying decoction for treating Graves' disease via thyroid cell apoptosis based on proteomics and molecular docking techniques

Ethnopharmacological relevanceFor numerous years, the Xiehuo Xiaoying decoction (XHXY), a traditional Chinese medicine formula, has demonstrated substantial promise in treating Graves' disease (GD) in clinical settings, showcasing significant potential. However, the therapeutic mechanism and efficacy material basis of XHXY remains obscure. Aim of the studyThis work aims to investigate the underlying mechanisms and to study the efficacy material basis of XHXY in anti-GD effect using a combination of TMT quantitative proteomics and molecular docking method. Materials and methodsGD model was initiated by administering Ad-TSH289. Subsequently, the mice underwent a four-week regimen that included oral gavage of XHXY at doses of 17 g/kg·d and 34 g/kg·d, along with intraperitoneal injections of Gentiopicroside (GPS). Utilizing the principles of pharmacological chemistry in traditional Chinese medicine, we employed high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-QTOF/MS) to discern prescribed prototype composition of XHXY in serum samples from mouse. TMT proteomics research provided evidence of XHXY's putative targets and important pathways in vivo. The binding activity of probable action targets and prototype composition was detected by molecular docking. Finally, Immunohistochemistry (IHC) and TUNEL staining were used to verify the mechanism of XHXY and GPS in anti-GD. ResultsXHXY and GPS alleviated GD by ameliorating the pathological changes and reducing thyroxine and TRAb levels. In mouse serum, a total of 31 prototypical XHXY ingredients were detected, and the majority of these components were from monarch and minister medicine. Proteomics study results indicated that the XHXY may mainly regulate targets including FAS-associated death domain protein (FADD), Apolipoprotein C-III, etc. and main pathways are Apoptosis, Cholesterol metabolism, TNF signalling pathway, etc. Strong binding activity of the prototypical active ingredient and GPS towards FADD, Caspase 8, and Caspase 3 was demonstrated by molecular docking. XHXY and its primary component, GPS, elevated the expression of FADD, Caspase 8, and Caspase 3, and enhance apoptosis in thyroid cells, as lastly validated by TUNEL and IHC staining. ConclusionsXHXY exhibits a favorable therapeutic effect in treating GD by promoting apoptosis in thyroid cells through the upregulation of FADD, Caspase 8, and Caspase 3 expression. And GPS is the main efficacy material basis for its therapeutic effect in anti-GD.

Mycobacterium vaccae alleviates allergic airway inflammation and airway hyper-responsiveness in asthmatic mice by altering intestinal microbiota.

Host immunity can influence the composition of the gut microbiota and consequently affect disease progression. Previously, we reported that a Mycobacterium vaccae vaccine could ameliorate allergic inflammation in asthmatic mice by regulating inflammatory immune processes. Here, we investigated the anti-inflammatory effects of M. vaccae on allergic asthma via gut microbiota modulation. An ovalbumin (OVA)-induced asthmatic murine model was established and treated with M. vaccae. Gut microbiota profiles were determined in 18 BALB/c mice using 16S rDNA gene sequencing and metabolomic profiling was performed using liquid chromatography quadrupole time-of-flight mass spectrometry. Mycobacterium vaccae alleviated airway hyper-reactivity and inflammatory infiltration in mice with OVA-induced allergic asthma. The microbiota of asthmatic mice is disrupted and that this can be reversed with M. vaccae. Additionally, a total of 24 differential metabolites were screened, and the abundance of PI(14:1(9Z)/18:0), a glycerophospholipid, was found to be correlated with macrophage numbers (r = 0.52, p = 0.039). These metabolites may affect chemokine (such as macrophage chemoattractant protein-1) concentrations in the serum, and ultimately affect pulmonary macrophage recruitment. Our data demonstrated that M. vaccae might alleviate airway inflammation and hyper-responsiveness in asthmatic mice by reversing imbalances in gut microbiota. These novel mechanistic insights are expected to pave the way for novel asthma therapeutic strategies.

An inulin-type fructan CP-A from Codonopsis pilosula alleviates TNBS-induced ulcerative colitis based on serum-untargeted metabolomics.

Ulcerative colitis (UC) is an inflammatory disease with abdominal pain, diarrhea, and bloody stool as the main symptoms. Several studies have confirmed that polysaccharides are effective against UC. It is commonly accepted that the traditional benefits of Radix Codonopsis can be attributed to its polysaccharide contents, and inulin-type fructan CP-A is the main active monomer in the polysaccharide components. Herein, we established a 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced UC rat model and lipopolysaccharide (LPS)-induced colonic epithelial cell model (NCM460) to investigate the effect of CP-A on UC. Untargeted metabolomics studies were conducted to identify differential metabolites using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS) and enrich metabolic pathways in rat serum. The in vivo assays demonstrated that CP-A reduces colonic macroscopic injury, disease activity index (DAI), histopathological score, interleukin (IL)-8, and tumor necrosis factor-α (TNF-α) levels, as well as the expression of intercellular adhesion molecules. On the other hand, CP-A increases IL-10 and transforming growth factor-β (TGF-β) levels. The in vitro experiments indicated that CP-A treatment could reduce nitric oxide (NO) and IL-1β after LPS stimulation. The metabolomics results suggested that CP-A therapy for UC may be related to the mammalian target of rapamycin (mTOR) signaling pathway. The in vitro and in vivo validation of the pathway showed similar results, indicating that CP-A alleviates UC by preventing the activation of mTOR/p70S6K signaling pathway. These findings offer a fresh approach to treating UC and a theoretical foundation for the future advancement of CP-A.NEW & NOTEWORTHY We report that an inulin-type fructan from Codonopsis pilosula CP-A exhibits a therapeutic effect on experimental colitis. Its mechanism may be to alleviate intestinal inflammation by preventing the activation of mammalian target of rapamycin (mTOR)/p70S6K signaling pathway. These findings offer a fresh approach to treating ulcerative colitis (UC) and a theoretical foundation for the future advancement of CP-A.

A breakthrough in periodontitis treatment: Revealing the pharmacodynamic substances and mechanisms of Kouqiangjie formula

Ethnopharmacology relevancePeriodontitis, a complex inflammatory disease, significantly affects people's lives. Traditional Chinese multi-herbal formulas, composed of various herbs, exhibit their therapeutic efficacy holistically. Kouqiangjie Formula (KQJF), comprising 12 herbs including Rhizoma smilacis glabrae, Polygonatum sibiricum Delar. ex Redoute, Taraxacum mongolicum Hand.-Mazz, etc., has been clinically proven to effectively treat periodontitis. However, the potential active substances conferring these effects and their mechanisms of action remain unclear. Aim of the studyThe current investigation endeavours to utilize Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-Q-TOF-MS), network pharmacology, and in vivo animal experiment confirmation to explore the plausible bioactive compounds and operational mechanisms underpinning KQJF's therapeutic impact on periodontitis. Materials and methodsUsing the UPLC-Q-TOF-MS technique, we deciphered the chemical constituents of KQJF. Network pharmacology was employed to earmark key bioactive elements, forecast principal targets, and operational pathways which were later substantiated through molecular docking. Experimental validations were carried out in a periodontitis animal model using a range of techniques, including micro-CT, H&E staining, qRT-PCR, and protein blotting procedures, providing comprehensive verification of our initial assumptions. ResultsUtilizing UPLC-Q-TOF-MS, we characterized 87 individual chemical constituents in KQJF. Network pharmacology revealed that 14 components, including senkyunolide A, glycycoumarin, licoflavonol, glycyrin, senkyunolide I, and senkyunolide H, form the key therapeutic basis of KQJF in targeting periodontitis. Significant targets and pathways were discerned as AKT1, MMP9, JUN, PTGS2, CASP3, TLR4, IL1β, BCL2, PPARG, and pathways such as the TNF signaling pathway, NF-κB signaling pathway, osteoclast differentiation, and Wnt signaling pathway. Molecular docking demonstrated robust binding activity between these crucial targets and the key active ingredients. In vivo experimentation corroborated that, compared with the model group, KQJF significantly ameliorated symptoms and micro-CT imaging parameters of periodontitis in the rat model, down-regulating the expression of AKT1, MMP9, JUN, PTGS2, CASP3, TLR4, and IL1β, while up-regulating the expression of BCL2 and PPARG. ConclusionIn summary, this study has pioneered a comprehensive exploration of the potential therapeutic constituents, targets, and mechanisms of KQJF for periodontitis treatment, adopting a synergistic strategy of “chemical component analysis-network pharmacology screening-in vivo animal experiment validation”. This provides experimental evidence for the clinical application of KQJF and further in-depth research. Additionally, it presents an effective strategy for the research of other Chinese herbal formulations.

A comparative metabolomics study on two fresh edible lilies for vegetable: Lilium brownii var. viridulum and Lilium davidii var. unicolor

Lily has been frequently used as food or medicine. Lilium brownii var.viridulum (L. brownii) and L. davidii var. unicolor are mainstream fresh edible species for vegetable. To investigate the differences between two varieties, free sugars, organic acids analysis and untargeted metabolomics were conducted. Thirty-one differential phytochemicals were annotated via ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry, which mainly enriched in amino acids and carbohydrates metabolic pathways. Seven phytochemicals, daidzin, isomaltulose, melibiose, melezitose, isorubijervosine, trigonelline and lycoperoside-D, were identified in lily for the first time. Compared with L. brownii, the contents of sugars and glutamine were higher, the bitter amino acids arginine, tyrosine, and alkaloids were lower in L. davidii var. unicolor. Meanwhile, the higher abundance of ferulic acid and p-coumaric acid presented sour rather than bitter taste in L. davidii var. unicolor. The results will provide new insights into the differences between L. brownii and L. davidii var. unicolor.

Study on the white frost formation mechanism during storage of Phyllanthus emblica Linn. fruit based on component analysis and spatial metabolomics

The Phyllanthus emblica Linn. fruit (PEF) is a well-known medicinal and food homologous item in tropical Southeast Asian. During the drying and storing processes, some PEF will grow white frost on its surface, which is typically taken as a sign of greater quality. However, the material basis and formation mechanism of white frost on PEF surfaces are currently unclear, and there is no sufficient evidence to support the correlation between white frost on PEF surfaces and their quality. In this paper, high-performance liquid chromatography (HPLC) was used to study the differences in active ingredient content of PEF medicinal materials with and without frost. The microstructure and elemental composition of white frost were studied using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). The Fourier transform infrared spectroscopy (FT-IR) was used to analysis the main functional groups in white frost. The ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) combined with UNIFI database, EDS and FT-IR results, and reference materials were used to identify the chemical composition of white frost. The exocarp of PEF before and after drying and storage was analyzed by spatial metabolomics using desorption electrospray ionization (DESI) mass spectrometry imaging system to reveal the formation mechanism of white frost on the surface of PEF. The results found that the PEF with frost have higher levels of active ingredients than those without frost. EDS and FT-IR results show that white frost is mainly composed of C, O, K elements, and contains a large number of phenolic hydroxyl, carboxyl etc. UPLC-Q-TOF-MS/MS results found that the main components of white frost were organic acids, fatty acids, and tannins, including quality markers such as gallic acid and ellagic acid etc. Spatial metabolomics research found that the white frost formation mechanism mainly involved in the ascorbate and aldarate metabolism, cutin, suberin and wax biosynthesis, citrate cycle (TCA cycle) and biosynthesis of unsaturated fatty acid. This study reveals the material basis, formation mechanism, and relationship between the surface white frost of PEF and the quality of medicinal materials, providing valuable information for the quality evaluation of PEF.

Aminoacylase efficiently hydrolyses fatty acid amino acid conjugates of Helicoverpa armigera potentially to increase the pool of glutamine

One of the most prevalent bioactive molecules present in the oral secretion (OS) of lepidopteran insects is fatty acid amino acid conjugates (FACs). Insect dietary components have influence on the synthesis and retaining the pool of FACs in the OS. We noted differential and diet-specific accumulation of FACs in the OS of Helicoverpa armigera by using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry. Interestingly, we identified FACs hydrolyzing enzyme aminoacylase (HaACY) in the OS of H. armigera through proteomic analysis. Next, we have cloned, expressed, and purified active recombinant HaACY in the bacterial system. Recombinant HaACY hydrolyzes all the six identified FACs in the OS of H. armigera larvae fed on host and non-host plants and releases respective fatty acid and glutamine. In these six FACs, fatty acid moieties vary while amino acid glutamine was common. Glutamine obtained upon hydrolysis of FACs by HaACY might serve as an amino acid pool for insect growth and development. To understand the substrate choices of HaACY, we chemically synthesized, purified, and characterized all the six FACs. Interestingly, rHaACY also shows hydrolysis of synthetic FACs into respective fatty acid and glutamine. Our results underline the importance of diet on accumulation of FACs and role of aminoacylase(s) in regulating the level of FACs and glutamine.

Bioprospecting of soil-borne microorganisms and chemical dereplication of their anti-microbial constituents with the aid of UPLC-QTOF-MS and molecular networking approach.

Due to the emergence of drug-resistant microorganisms, the search for broad-spectrum antimicrobial compounds has become extremely crucial. Natural sources like plants and soils have been explored for diverse metabolites with antimicrobial properties. This study aimed to identify microorganisms from agricultural soils exhibiting antimicrobial effects against known human pathogens, and to highlight the chemical space of the responsible compounds through the computational metabolomics-based bioprospecting approach. Herein, bacteria were extracted from soil samples and their antimicrobial potential was measured via the agar well diffusion method. Methanolic extracts from the active bacteria were analyzed using the liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) technique, and the subsequent data was further analyzed through molecular networking approach which aided in identification of potential anti-microbial compounds. Furthermore, 16S rRNA gene sequencing enabled identification of the active bacterial isolates, where isolate 1 and 2 were identified as strains of Bacillus pumilus, whilst isolate 3 was found to be Bacillus subtilis. Interestingly, isolate 3 (Bacillus subtilis) displayed wide-ranging antimicrobial activity against the tested human pathogens. Molecular networking revealed the presence of Diketopiperazine compounds such as cyclo (D-Pro-D-Leu), cyclo (L-Tyr-L-Pro), cyclo (L-Pro-D-Phe), and cyclo (L-Pro-L-Val), alongside Surfactin C, Surfactin B, Pumilacidin E, and Isarrin D in the Bacillus strains as the main anti-microbial compounds. The application of the molecular networking approach represents an innovation in the field of bio-guided bioprospection of microorganisms and has proved to be an effective and feasible towards unearthing potent antimicrobial compounds. Additionally, the (computational metabolomics-based) approach accelerates the discovery of bioactive compounds and isolation of strains which offer a promising avenue for discovering new clinical antimicrobials. Finally, soil microbial flora could serve an alternative source of anti-microbial compounds which can assist in the fight against emergence of multi-drug resistance bacterial pathogens.

The Beneficial Effects of Dietary Astragali Radix Are Related to the Regulation of Gut Microbiota and Its Metabolites.

Astragali Radix (AR) or its extract has been used as an herbal medicine and dietary supplement in China, Europe, and the United States. The gut microbiota could provide new insights for exploring dietary supplements' underlying mechanism on organisms. However, no reports have focused on the regulatory effect of AR on the gut microbiota as a dietary supplement. In this study, healthy ICR mice of either sex were divided into AR and control (CON) groups and given AR water extract (4.55 mg/kg·day-1) or saline by gavage for 14 days, respectively. Then 16S rRNA gene sequencing and ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry-based fecal metabolomics were integrated to investigate the benefits of dietary AR. Weighted gene coexpression network analysis was also introduced to investigate the metabolites with highly synergistic changes. AR supplementation influenced the structure of intestinal microflora, especially enriching short-chain fatty acid-producing bacteria g_Coprobacillus, g_Prevotella, and g_Parabacteroides. AR also significantly altered the fecal metabolome, mainly related to amino acid metabolism, nucleotide metabolism, and bile acid (BA) metabolism. Moreover, the increased secondary BAs and BA-sulfates might closely relate to intestinal microflora. These findings provide valuable insights for future research of dietary AR as a functional food.

Exposure of Helicobacter pylori to clarithromycin in vitro resulting in the development of resistance and triggers metabolic reprogramming associated with virulence and pathogenicity.

In H. pylori infection, antibiotic-resistance is one of the most common causes of treatment failure. Bacterial metabolic activities, such as energy production, bacterial growth, cell wall construction, and cell-cell communication, all play important roles in antimicrobial resistance mechanisms. Identification of microbial metabolites may result in the discovery of novel antimicrobial therapeutic targets and treatments. The purpose of this work is to assess H. pylori metabolomic reprogramming in order to reveal the underlying mechanisms associated with the development of clarithromycin resistance. Previously, four H. pylori isolates were induced to become resistant to clarithromycin in vitro by incrementally increasing the concentrations of clarithromycin. Bacterial metabolites were extracted using the Bligh and Dyer technique and analyzed using metabolomic fingerprinting based on Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (LC-Q-ToF-MS). The data was processed and analyzed using the MassHunter Qualitative Analysis and Mass Profiler Professional software. In parental sensitivity (S), breakpoint isolates (B), and induced resistance isolates (R) H. pylori isolates, 982 metabolites were found. Furthermore, based on accurate mass, isotope ratios, abundances, and spacing, 292 metabolites matched the metabolites in the Agilent METLIN precise Mass-Personal Metabolite Database and Library (AM-PCDL). Several metabolites associated with bacterial virulence, pathogenicity, survival, and proliferation (L-leucine, Pyridoxone [Vitamine B6], D-Mannitol, Sphingolipids, Indoleacrylic acid, Dulcitol, and D-Proline) were found to be elevated in generated resistant H. pylori isolates when compared to parental sensitive isolates. The elevated metabolites could be part of antibiotics resistance mechanisms. Understanding the fundamental metabolome changes in the course of progressing from clarithromycin-sensitive to breakpoint to resistant in H. pylori clinical isolates may be a promising strategy for discovering novel alternatives therapeutic targets.

ERCC1 which affects lipids metabolism and actin dynamics in coal workers' pneumoconiosis is a candidate biomarker for early warning and diagnosis.

The single-nucleotide polymorphisms of genes related to DNA damage repair and inflammasomes and mutated gene expression in coal workers' pneumoconiosis (CWP) were analysed to identify the risk factors of CWP and potential biomarkers for early warning and diagnosis. Further, mutated gene pathways were analysed based on proteome and metabolome. Han Chinese male subjects were randomly selected and divided into 4 or 5 groups according to the process of CWP. MassARRAY was used to sequence single-nucleotide polymorphism genotypes. Mutated gene expression in plasma was tested using enzyme-linked immunosorbent assay (ELISA). Odds ratios (ORs) and receiver operating characteristic curves (ROC) were calculated. The serum different proteins and metabolites were identified by Ultra Performance Liquid Chromatography Quadrupole time of flight/Mass Spectrum (UPLC-Q-TOF/MS) and analysed using bioinformation software. As CWP progressed, the CC and CA genotypes of ERCC1 rs3212986 decreased and increased significantly, respectively. AA (OR = 3.016) and CA (OR = 2.130) genotypes were identified as risk factors for stage II. ERCC1 significantly decreased in processing of CWP. The cutoff value of ERCC1 was 5.265 pg/ml, with a sensitivity of 90.0% and specificity of 86.7%. ERCC1 had an indirect interaction with activator protein-1 and insulin and its pathways were mainly made with molecules related to lipid metabolism and actin dynamics. ERCC1 is a candidate biomarker for detection and precise intervention in CWP. If it reaches the threshold, workers will change other jobs in time and will not develop and diagnose as pneumoconiosis and will help the employers spend less money. Meanwhile, the signal molecules of ERCC1 pathway could be as a candidate target for drug discovery.

Integration of Network Pharmacology and Experimental Validation to Explore Jixueteng - Yinyanghuo Herb Pair Alleviate Cisplatin-Induced Myelosuppression.

Jixueteng, the vine of the bush Spatholobus suberectus Dunn., is widely used to treat irregular menstruation and arthralgia. Yinyanghuo, the aboveground part of the plant Epimedium brevicornum Maxim., has the function of warming the kidney to invigorate yang. This research aimed to investigate the effects and mechanisms of the Jixueteng and Yinyanghuo herbal pair (JYHP) on cisplatin-induced myelosuppression in a mice model. Firstly, ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) screened 15 effective compounds of JYHP decoction. Network pharmacology enriched 10 genes which may play a role by inhibiting the apoptosis of bone marrow (BM) cells. Then, a myelosuppression C57BL/6 mice model was induced by intraperitoneal (i.p.) injection of cis-Diaminodichloroplatinum (cisplatin, CDDP) and followed by the intragastric (i.g.) administration of JYHP decoction. The efficacy was evaluated by blood cell count, reticulocyte count, and histopathological analysis of bone marrow and spleen. Through the vivo experiments, we found the timing of JYHP administration affected the effect of drug administration, JYHP had a better therapeutical effect rather than a preventive effect. JYHP obviously recovered the hematopoietic function of bone marrow from the peripheral blood cell test and pathological staining. Flow cytometry data showed JYHP decreased the apoptosis rate of BM cells and the western blotting showed JYHP downregulated the cleaved Caspase-3/Caspase-3 ratios through RAS/MEK/ERK pathway. In conclusion, JYHP alleviated CDDP-induced myelosuppression by inhibiting the apoptosis of BM cells through RAS/MEK/ERK pathway and the optimal timing of JYHP administration was after CDDP administration.

Method development and optimization for dispersive liquid–liquid microextraction factors using response surface methodology with desirability function for the ultra–high performance liquid chromatography quadrupole time of flight mass spectrometry determination of organic contaminants in water samples: Risk and greenness assessment.

Method development and optimization for dispersive liquid–liquid microextraction factors using response surface methodology with desirability function for the ultra–high performance liquid chromatography quadrupole time of flight mass spectrometry determination of organic contaminants in water samples: Risk and greenness assessment.

A low-volume LC/MS method for highly sensitive monitoring of phenoxymethylpenicillin, benzylpenicillin, and probenecid in human serum.

Background: The optimization of antimicrobial dosing plays a crucial role in improving the likelihood of achieving therapeutic success while reducing the risks associated with toxicity and antimicrobial resistance. Probenecid has shown significant potential in enhancing the serum exposure of phenoxymethylpenicillin, thereby allowing for lower doses of phenoxymethylpenicillin to achieve similar pharmacokinetic/pharmacodynamic (PK/PD) targets. We developed a triple quadrupole liquid chromatography mass spectrometry (TQ LC/MS) analysis of, phenoxymethylpenicillin, benzylpenicillin and probenecid using benzylpenicillin-d7 and probenecid-d14 as IS in single low-volumes of human serum, with improved limit of quantification to support therapeutic drug monitoring. Methods: Sample clean-up was performed by protein precipitation using acetonitrile. Reverse phase chromatography was performed using TQ LC/MS. The mobile phase consisted of 55% methanol in water + 0.1% formic acid, with a flow rate of 0.4 mL min-1. Antibiotic stability was assessed at different temperatures. Results: Chromatographic separation was achieved within 2 minutes, allowing simultaneous measurement of phenoxymethylpenicillin, benzylpenicillin and probenecid in a single 15 μL blood sample. Validation indicated linearity over the range 0.0015-10 mg L-1, with accuracy of 96-102% and a LLOQ of 0.01 mg L-1. All drugs demonstrated good stability under different storage conditions. Conclusion: The developed method is simple, rapid, accurate and clinically applicable for the quantification of phenoxymethylpenicillin, benzylpenicillin and probenecid in tandem.