Quadrupole Orbitrap Mass Spectrometry Research Articles

Effect of Hong Qu on the flavor and quality of Hakka yellow rice wine (Huangjiu) produced in Southern China

This study aimed to investigate effect of Hong Qu on flavor and quality of Hakka yellow rice wine (Hakka Huangjiu). Volatile components of three kinds of Hakka Huangjiu with or without adding Hong Qu were analyzed by solid phase microextraction-gas chromatography-mass spectrometry. Their non-volatile components were also detected by liquid chromatography quadrupole Orbitrap mass spectrometry (LC-Q-Orbitrap-MS). Results indicated that there were great differences in flavor components among three wines. Among them, the main difference components between traditional Hakka Huangjiu and Hakka Huangjiu without adding Hong Qu were isoamyl alcohol, phenylethanol, isobutyl acetate, isobutyl alcohol, ethyl palmitate and ethyl decanoate. In Hakka Huangjiu, esters were the most volatile components, followed by alcohols, while acids were less. Under NEG and POS modes of LC-Q-Orbitrap-MS, a total of 765 and 1244 components were detected in three kinds of Hakka Huangjiu, respectively. Among 217 kinds of polypeptides detected, the content of 160 kinds of polypeptides in Hakka Huangjiu adding only with Hong Qu was higher than that of traditional Hakka Huangjiu and Hakka Huangjiu without adding Hong Qu. Amino acids and polypeptides contents in Hakka Huangjiu brewed only with Hong Qu and traditional Hakka Huangjiu co-fermented with Hong Qu were relatively high.

Spectrum-effect relationship between ultra-high-performance liquid chromatography fingerprints and antioxidant activities of Lophatherum gracile Brongn.

Lophatherum gracile Brongn. is a medicinal and edible plant resource as well as a natural additive in the functional food market. To better understand its characteristics and efficacy, a method combining chromatographic fingerprints and antioxidant activity was proposed. A total of 21 common peaks were confirmed from liquid chromatography fingerprints and were identified as 14 flavonoids and 7 phenolic acids by ultra‐high‐performance liquid chromatography (UHPLC) coupled with quadrupole Orbitrap mass spectrometry (Q‐Orbitrap/MS). Their antioxidant activities were evaluated by 1,1‐diphenyl‐2‐trinitrophenylhydrazine (DPPH), 2,2'‐diazide‐bis (3‐ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS), and ferric reducing antioxidant power (FRAP) assay. The results showed that all of the test samples had moderate to high antioxidant effects, with IC50 values ranging from 5.2 to 16.1 mg/ml and 1.2 to 2.8 mg/ml for DPPH and ABTS assays, and the FeSO4 concentrations of 1.84–4.20 mmol/L for the FRAP assay. The spectrum–effect relationship between UHPLC fingerprints and antioxidant activity was investigated through Pearson correlation analysis and Grey relational analysis (GRA) to identify the antioxidant constitutes in Lophatherum gracile Brongn. The results showed that 11 compounds were greatly associated with the antioxidant activity with a correlation degree >0.80, which can be used as the quality marker of Lophatherum gracile Brongn.

Molecular mechanism of Mare Nectaris and magnetic field on the formation of ethyl carbamate during 19 years aging of Feng-flavor Baijiu

Ethyl carbamate (EC) is carcinogen occurring naturally in fermented foods, while the EC formation pattern in Feng-flavor Baijiu during Mare Nectaris storage and magnetic field treatment remains controversial. In this work, variation of EC in Mare Nectaris and magnetic field were investigated for the first time through ultra high performance liquid chromatography quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap). Quantification results revealed that EC decreased significantly in the stage of 3–9 years and kept at 12.4 μg L−1 after 10 years of aging. Arginine succinate synthase (ASS) and urease were deemed as vital factors for EC decomposition. Degradation effetc of EC in 250 mT is simillar to that of EC in Baijiu stored in Mare Nectaris for 8 years. This is due to that aging process was accelerated by magnetic field and the content of total acid in Baijiu was increased, creating a favorable environment for decomposition of EC and urea.

Biomass magnetic porous carbon prepared from mangosteen shell for the preconcentration of three bisphenols from beverages followed by liquid chromatography with quadrupole-Orbitrap high-resolution mass spectrometry determination.

Natural biomass magnetic porous carbon was successfully prepared via a cost-effective and green route using mangosteen shells as raw material. The prepared magnetic porous carbon was used as a magnetic solid-phase extraction adsorbent for bisphenols enrichment from beverages followed by high-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry. Parameters affecting extraction efficiency including sample solution pH, adsorbent amount, extraction time, eluent type, and volume were optimized. Results showed that biomass magnetic porous carbon had excellent adsorption properties for bisphenols due to its large specific surface area and abundant functional groups, which could form hydrogen bonding and π-π stacking with bisphenols. The enrichment factor of 3 bisphenols was in the range of 15-19. Under optimum conditions, favorable linearity for all analytes was obtained with correlation coefficients higher than 0.998. Recoveries of spiked samples were in the range of 88.5-105.1% with a relative standard deviation of 3.4-5.5%. These results demonstrated that magnetic porous carbon may be a promising adsorbent for the enrichment of aromatic compounds.

Molecular mechanism of high pressure shear grinding on Feng-flavour Chinese Baijiu ageing

Ultrahigh-pressure homogenization is a powerful technique for accelerating Baijiu ageing. In this study, the molecular mechanism of artificial ageing was thoroughly investigated utilizing foodomics combined with a mass spectrum information optimization program. Various characterizations of compounds in the ageing process were performed through ultra high performance liquid chromatography quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap) based colloid theoretics. High-pressure shear grinding provides a high-pressure and high-temperature environment for Baijiu, accelerating the formation of flavour compounds and the decomposition of odour and harmful substances, thus improving the sensory characteristics of Feng-flavour Baijiu and shortening the ageing time. A total of 2069 substances were identified according to fragment spectra and Bayesian network model-based peak distribution. Molecules with VIP > 1, including 14 esters, 10 acids, 4 ketones, 3 aldehydes, 2 alcohols and 2 phenols, were quantified as potential markers of chemometric analysis. A novel prediction model for Baijiu ageing degree was furnished, and ageing years of Feng-flavour Chinese Baijiu were accurately predicted by factor analysis with KMO 0.784 and a polynomial regression model with r 0.94. The results demonstrated that in 400 bar, fresh distilled Baijiu could reach a natural ageing effect of 6.43 years, which was consistent with the sensory evaluation.

Quantification, identification and comparison of oligopeptides on five tea categories with different fermentation degree by Kjeldahl method and ultra-high performance liquid chromatography coupled with quadrupole-orbitrap ultra-high resolution mass spectrometry

Peptides with different lengths or amino sequences could have specific tastes or bio-activities. So far, either the quantity or pattern differences of peptide among various of teas were unknown. Here, firstly, we developed a method for tea oligopeptide quantification and made comparison of their contents. Secondly, we applied ultra-high performance liquid chromatography coupled with quadrupole-orbitrap ultra-high resolution mass spectrometry (UHPLC-Quadrupole-Orbitrap-UHRMS) to sequence oligopeptides. As a result, the total amount of oligopeptides in white tea and dark tea were higher, followed by black tea and green tea, finally with oolong tea. It suggested that withering which undergoes with endogenous protease and post-fermented that undergoes with a participation of exotic micro-organisms were key in oligopeptide enrichment. Thirdly, a total of 902 abundant identified peptides, most of which were tripeptide, tetrapeptide, pentapeptide, and hexapeptide were screened against several existing peptide databases. There were a series of taste peptides and bio-active peptides existing.

Protective effects on ovalbumin-induced mouse asthma models and qualitative and quantitative analysis of multiple compounds in Gerberae Piloselloidis Herba.

Gerberae Piloselloidis Herba is widely used to treat cough and asthma in China. However, its effects on allergic asthma as related to its chemical compositions have not been fully elucidated, and there is a scarcity of methods to determine multi-component contents for quality control. In this study, protective effects of Gerberae Piloselloidis Herba on ovalbumin-induced asthma models were investigated, while qualitative and quantitative analyses of multiple constituents in Gerberae Piloselloidis Herba were conducted by using an ultrahigh-performance liquid chromatography-Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry and an ultrahigh-performance liquid chromatography-photodiode array detection. The results showed that Gerberae Piloselloidis Herba could significantly mitigate asthma symptoms, reduce eosinophils counts in the bronchoalveolar lavage fluid, as well as decrease IgE, IL-5, and IL-13 concentration, and inflammatory cellular infiltration in lung tissues. A total of 51 compounds were tentatively identified, in which the content of 10 representative compounds was determined in 24 batches of Gerberae Piloselloidis Herba by using an ultrahigh-performance liquid chromatography method with good linearity, precision, repeatability, accuracy, and stability. This research presents a comprehensive strategy combining biological activity evaluation with chemical profiling, providing a useful and comprehensive reference for further application and quality control of Gerberae Piloselloidis Herba.

Comprehensive Study on the Chemical Profile and Anti-Tumor Activity of Secondary Metabolites Produced by Aspergillus niger

Objective: Chemical investigation of the extract of sponge-derived fungus, Aspergillus niger, was performed by liquid chromatography coupled with QExactive mass spectrometry for the first time. Methods: A total number of 444 constituents were detected, 288 of which were identified positively or tentatively by the comprehensive utilization of accurate molecular weight and fragmentation information acquired from quadrupole orbitrap mass spectrometry. The identified compounds were divided into several types, namely, organic acid, alkaloid, saccharide, amino acid and cyclopeptide, terpenoid, polyketone, phenylpropanoid and other types of compounds. Systematic diagnostic ions and featured fragment patterns were summarized for each type, based on which 8 novel compounds belonging to the same type were characterized. Results: This work provided a rapid approach for the research of micro constituents in a complex analyte. Furthermore, the anti-tumor activity of the extract was evaluated on two different cell lines-Bel-7402 and Hela-S3 in vitro. The tumor-inhibitory effect of the Aspergillus niger extract was confirmed and may be mainly derived from its pro-apoptotic action. Moreover, the extract exerted more significant cytotoxicity in Bel-7402 cells than Hela-S3 cells, indicative of its selectivity on specific tumor cells. Conclusion: The evidence suggested that the Aspergillus niger extract may potentially serve as a remedy for the prevention and therapy of hepatic and breast carcinoma.

Discovery and Molecular Elucidation of the Anti-Influenza Material Basis of Banlangen Granules Based on Biological Activities and Ultra-High Performance Liquid Chromatography Coupled with Quadrupole-Orbitrap Mass Spectrometry

Discovery and Molecular Elucidation of the Anti-Influenza Material Basis of Banlangen Granules Based on Biological Activities and Ultra-High Performance Liquid Chromatography Coupled with Quadrupole-Orbitrap Mass Spectrometry

Identification of the Metabolites of Scutebarbatine A in Rat Plasma, Bile, Urine, and feces by Using Ultra-high-performance Liquid Chromatography Coupled with Q Exactive Hybrid Quadrupole-orbitrap High-resolution Mass Spectrometry.

Scutebarbatine A is a new neo-Clerodane Diterpenoid Alkaloids from Scutellaria barbata, which has many pharmacological effects, such as anti-tumor, antibacterial, and anti-inflammatory. However, there are no studies on the metabolism of Scutebarbatine A. The objective of this study is to explore the metabolism of Scutebarbatine A in the body, the bile, plasma, urine, and feces samples of rats after administration were investigated. The biological samples were investigated using ultra-high-performance liquid chromatography coupled with Q Exactive hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap-HRMS). A total of 20 metabolites were identified: 16 phase I metabolites and 4 phase II metabolites. The main metabolic pathways were hydrolysis, oxidation, hydrogenation, dehydration, and combination with sulfate. This study further elucidates the metabolism of Scutebarbatine A in rats and provides a reference for the study of its pharmacodynamic material basis and pharmacological mechanism.

Metabolomics based plasma biomarkers for diagnosis of oral squamous cell carcinoma and oral erosive lichen planus.

Backgrounds: To identify diagnostic biomarkers for differentiating oral squamous cell carcinoma (OSCC) from oral erosive lichen planus (OELP) and investigate potential biomarkers associated with malignant transformation.Methods: In this study, 72 patients with OSCC, 75 patients with OELP subjects were recruited. Their plasma samples were analyzed by ultra-high-performance liquid chromatography quadrupole-Orbitrap high-resolution accurate mass spectrometry, (UHPLC/Q-Orbitrap HRMS). Principal component analysis, orthogonal partial least square discrimination analysis, t-test analysis and false discovery rate were used to identify different metabolites in patients with OSCC and OELP. The metabolic pathway analysis was performed by MetaboAnalyst. To further screen and identify the biomarkers of OSCC and establish a diagnostic panel, binary logistic regression analysis and receiver operating characteristic analysis were used. The data were then combined with blood samples from healthy individuals for mass spectrometry analysis to obtain biomarkers related to malignant transformation.Results: A total of 20 kinds of endogenous metabolites were identified from plasma samples of OSCC patients and OELP patients. Metabolic pathway analysis showed that the biomarkers associated with OSCC were closely related to cholic acid metabolism and amino acid metabolism. Finally, a diagnostic panel composed of decanoylcarnitine, cysteine and cholic acid was established. This diagnostic panel had good diagnostic efficiency with the AUC=0.998. Other metabolites including uridine, taurine, glutamate, citric acid and LysoPC(18:1) were identified to be general biomarkers for malignant transformation of OELP.Conclusion: Biomarkers based on plasma metabolomics are of great significance for the prediction of malignant transformation of OELP and early diagnosis of OSCC.

Glabrol impurity exacerbates glabridin toxicity in zebrafish embryos by increasing myofibril disorganization

Ethnopharmacological relevanceGlabridin, extracted from Glycyrrhiza glabra L., is widely used for the treatment of hyperpigmentation because of its anti-inflammatory and antioxidant activities and its ability to inhibit melanin synthesis. This led to the strict regulation of its quality and safety. However, traditional quality control methods used for plant extracts cannot reflect the product quality owing to multiple unknown impurities, which necessitates the further analysis of impurities. Aim of the studyThe study identified the toxic impurities of glabridin and their toxicological mechanism. Materials and methodsIn total, 10 glabridin samples from different sources were quantified using high-performance liquid chromatography. Sample toxicities were evaluated using zebrafish and cell models. To identify impurities, samples with different toxicity were analyzed by ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry. The toxicity of related impurities was verified in the zebrafish model. Phalloidin stain was used to evaluate subtle changes in myofibril alignment. ResultsAlthough glabridin content in the samples was similar, there were significant differences in toxicity. The results were verified using four different mammalian cell lines. Higher contents of glabrone and glabrol were identified in the sample with the highest toxicity. In the zebrafish model, the addition of glabrol reduced the LC50 of glabridin to 9.224, 6.229, and 5.370 μM at 48, 72, and 96 h post-fertilization, respectively, whereas glabrone did not have any toxic effect. Phalloidin staining indicated that a glabrol impurity exacerbates the myotoxicity of glabridin in zebrafish embryos. ConclusionGlabrol, but not glabrone, was identified as a key impurity that increased glabridin toxicity. This finding indicates that controlling glabrol content is necessary during glabridin product production.

Banana-peel-derived magnetic porous carbon as effective adsorbent for the enrichment of six bisphenols from beverage and water samples

Functionalized magnetic nanoporous carbon derived from banana peel was synthesized through carbonization, porogenesis, carboxylation and magnetization using banana peel and was successfully used as a magnetic solid phase extraction (MSPE) material for the enrichment of six bisphenols (BPs) from beverage and water samples. After the optimization of MSPE process, the enrichment factors of six target analytes were in the range of 74–112 for water samples, and 15–22 for beverage samples. Then, high-performance liquid chromatography–quadrupole-Orbitrap high-resolution mass spectrometry (HPLC–Q Orbitrap-HRMS) was used for the separation and determination of the target analytes. Results showed that the extraction recoveries for 6 BPs were in the range of 71.9–108.4% with an RSD of 2.5–7.5% (n = 6). These results demonstrated that the as-prepared material could efficiently enrich some aromatic compounds and the proposed method is reliable and robust for the determination of BPs in water and beverage samples.

Revealing the Toxicity-Enhancing Essence of Glycyrrhiza on Genkwa Flos Based on Ultra-high-performance Liquid Chromatography Coupled With Quadrupole-Orbitrap High-Resolution Mass Spectrometry and Self-Assembled Supramolecular Technology.

Researchers often focus on the mechanisms of synergistic agents, a few explore drug combinations that enhance toxicity, while few have studied the internal mechanism of compatibility enhancement in chemical level. Herein, we present a comprehensive analysis based on ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) and a self-assembled supramolecular strategy, which reveals the toxicity-enhancing essence of glycyrrhizic acid originated in licorice when combined with Genkwa Flos. Through this method, we discovered the toxicity was enhanced through the formation of a supramolecular complex from Genkwa Flos/glycyrrhizic acid. The morphology and size distribution of the self-assembled nanoparticles were characterized by scanning electron microscopy and dynamic light scattering Furthermore, a total of 58 constituents (eight diterpenoids, 35 flavonoids, five phenylpropanoids, four nucleosides, two amino acids, and four other compounds) consisted from the supramolecular complex were identified through accurate-mass measurements in full-scan MS/data-dependent MS/MS mode. Based on the hydrophobic interaction of glycyrrhizic acid with yuanhuacine (one of main ingredients from Genkwa Flos), the supramolecular self-assembly mechanism was revealed with proton nuclear magnetic resonance (1H-NMR) and NOESY 2D NMR. The toxicity of Genkwa Flos and Genkwa Flos/glycyrrhizic acid supramolecular complex were compared through in vitro studies on L-02 cells using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; and 4′,6-diamidino-2-phenylindole (DAPI) staining was performed to further confirm the enhancement inhibition of Genkwa Flos/glycyrrhizic acid supramolecular complex than Genkwa Flos. This study provides fundamental scientific evidence of the formation of a self-assembled phytochemical supramolecular when Genkwa Flos and glycyrrhizic acid are combined, enabling to understand their clinical incompatibility and contraindication.

Urinary non-targeted toxicokinetics and metabolic fingerprinting of exposure to 3-monochloropropane-1,2-diol and glycidol from refined edible oils

The widespread presence of 3-monochloropropane-1,2-diol (3-MCPD) and glycidol in refined edible oils have raised food industrial and public health concerns, but their specific biomarkers of exposure and urinary metabolic pathways indicating nephrotoxicity remain largely unknown. Here, we unraveled the in vivo biotransformation of these two contaminants and revealed how they affect metabolic pathways in rats. Urine metabolomes in rats administered with glycidol or 3-MCPD were investigated using ultra-high performance liquid chromatography combined with a quadrupole-orbitrap high-resolution mass spectrometry. Compared to the currently acknowledged metabolite which is only 2,3-dihydroxypropyl mercapturic acid, we identified 8 and 4 new specific exposure biomarkers of glycidol and 3-MCPD, respectively, via mapping the glyceryl polymerization and glutathione and sulfur conjugation. The changes of metabolites in the surrounding metabolic network were investigated to further gain insight into their metabolic fates. Exposure to glycidol up-regulated citrate, isocitrate, ketoglutarate, malate, and pyruvate in the tricarboxylic acid cycle and glycolysis pathways, while 3-MCPD intake down-regulated these signal molecules in both pathways. Nonetheless, L-cysteine, proline, and arginine were significantly decreased by the effect of either glycidol or 3-MCPD. Our findings first map the urinary metabolomics of both contaminants from edible oils and advance the omics-level recognition for their observational health hazards.

Comparative analysis of absorbed ingredients and metabolites, and pharmacokinetic studies of Zhimu-Huangbai herb pair in the plasma of normal and type 2 diabetes mellitus rats by UHPLC-linear trap quadrupole-orbitrap MS and LC-MS/MS.

A new rapid ultra-high-performance liquid chromatography coupled with linear trap quadrupole orbitrap mass spectrometry method was established for the qualitative analysis of absorbed ingredients and metabolites of Zhimu-Huangbai herb pair, which is used to treat type 2 diabetes mellitus. A total of 16 absorbed ingredients and 11 metabolites were identified in normal and type 2 diabetes mellitus rats, respectively. Such findings indicated that the diabetic model had no effect on the type of components in plasma. Seven absorbed ingredients and 11 metabolites were first identified after the oral administration of Zhimu-Huangbai herb pair. Thereafter, ultra-high-performance liquid chromatography coupled with linear trap quadrupole orbitrap mass spectrometry and liquid chromatography-API4000+ triple quadrupole mass spectrometer methods were established and validated for pharmacokinetic comparative studies of seven major bioactive components in normal and type 2 diabetes mellitus rats. Partial pharmacokinetic parameters in the plasma of type 2 diabetes mellitus rats were significantly different from those in normal rats. To our knowledge, this is the first comparison of absorbed ingredients and metabolites of Zhimu-Huangbai herb pair, and its use in pharmacokinetic studies between normal and type 2 diabetes mellitus rats. Ultimately, our findings provide insights into the clinical usage of Zhimu-Huangbai herb pair.

Comprehensive analysis of Sini decoction and investigation of acid-base self-assembled complexes using cold spray ionization mass spectrometry

Decoctions are a staple of traditional Chinese medicine. This study reports on a comprehensive analysis of the chemical compositions of both the supernatant and naturally-occurring precipitate of Sini decoction using ultra-high-performance liquid chromatography (UHPLC) hyphenated with quadrupole-Orbitrap high-resolution mass spectrometry (Q-Orbitrap HRMS). A total of 99 compounds (40 flavonoids, 34 alkaloids, 7 terpenoids, 3 phenolic acids and 15 other compounds) were tentatively identified in the full-scan MS/data-dependent MS/MS (full-scan MS1/dd-MS2) acquisition mode. Moreover, cold spray ionization-mass spectrometry (CSI-MS) was used to explore the presence of self-assembled complexes between phytochemical ingredients. As a proof-of-concept demonstration, the complexation between one acidic compound (glycyrrhizic acid) and three basic compounds (aconitine, hypaconitine, and mesaconitine) was characterized. This study may present inspiring scientific evidence for the formation of naturally-occurring precipitate, enabling a better understanding of Sini decoction and its therapeutic relevance.

Suspect and non-target screening of chemicals in clothing textiles by reversed-phase liquid chromatography/hybrid quadrupole-Orbitrap mass spectrometry

The global manufacturing of clothing is usually composed of multistep processes, which include a large number of chemicals. However, there is generally no information regarding the chemical content remaining in the finished clothes. Clothes in close and prolonged skin contact may thus be a significant source of daily human exposure to hazardous compounds depending on their ability to migrate from the textiles and be absorbed by the skin. In the present study, twenty-four imported garments on the Swedish market were investigated with respect to their content of organic compounds, using a screening workflow. Reversed-phase liquid chromatography coupled to electrospray ionization/high-resolution mass spectrometry was used for both suspect and non-target screening. The most frequently detected compound was benzothiazole followed by quinoline. Nitroanilines with suspected mutagenic and possible skin sensitization properties, and quinoline, a carcinogenic compound, were among the compounds occurring at the highest concentrations. In some garments, the level of quinoline was estimated to be close to or higher than 50,000 ng/g, the limit set by the REACH regulation. Other detected compounds were acridine, benzotriazoles, benzothiazoles, phthalates, nitrophenols, and organophosphates. Several of the identified compounds have logP and molecular weight values enabling skin uptake. This pilot study indicates which chemicals and compound classes should be prioritized for future quantitative surveys and control of the chemical content in clothing as well as research on skin transfer, skin absorption, and systemic exposure. The results also show that the current control and prevention from chemicals in imported garments on the Swedish market is insufficient.Graphical abstract

Digestion-Promoting Effects and Mechanisms of Dashanzha Pill Based on Raw and Charred Crataegi Fructus.

Emerging evidence suggests that a high-fat diet (HFD) can influence endoplasmic reticulum (ER) stress and gut microbiota. Crataegi Fructus is a traditional Chinese herb widely used in formulas for dyspepsia, with Dashanzha Pill composed of raw Crataegi Fructus (DR) being a representative drug. Processing products of Crataegi Fructus, however, have a stronger pro-digestive effect, and we hypothesized that Dashanzha Pill composed of charred Crataegi Fructus (DC) is more effective. We found that the contents of glucose 1-phosphate and luteolin in DR and DC were substantially different via ultra-high performance liquid chromatography-hybrid quadrupole-Orbitrap high-resolution mass spectrometry. DC outperformed DR in improving histopathological changes, increasing gastrin and motilin, and decreasing vasoactive intestinal peptides in rats with HFD induced dyspepsia. Fecal microbiota analysis revealed that DC could restore the disturbed intestinal microbiota composition, including that of Bacteroides, Akkermansia, and Intestinimonas to normal levels. Furthermore, DC significantly reduced the mRNA and protein levels of glucose-regulated protein 78, protein kinase R-like ER kinase, and eukaryotic initiation factor 2α. Taken together, DC outperformed DR in relieving dyspepsia by regulating gut microbiota and alleviating ER stress.

Metabolic study of cafestol using in silico approach, zebrafish water tank experiments and liquid chromatography high-resolution mass spectrometry analyses

Coffee is one of the most consumed beverages worldwide. Cafestol is an endogenous coffee diterpene present in raw coffee beans and also found in hot beverages, with several biological activities. However, there is still little information on this molecule after ingestion of coffee infusion.Zebrafish (Danio rerio) is a promising in vivo model for metabolic studies due to the annotation of mammalian orthologs to encode enzymes related to drug metabolism. Experiments using Zebrafish Water Tank (ZWT) model produce more significant number of metabolites for molecular investigation in a cleaner matrix than other classical models, such as purified hepatocytes.This work aimed to investigate the biotransformation of cafestol by the ZWT model using ultra-performance liquid chromatography coupled to hybrid quadrupole-orbitrap high-resolution mass spectrometry equipped with electrospray ionization (UPLC-HRMS) supported by in silico approach using SMARTCyp, Way2Drug and XenoSite Softwares.Twenty-five metabolites of cafestol were proposed by in silico analysis, in which 5 phase I metabolites were confirmed in the ZWT by UPLC and MS/HRMS investigation: 6-hydroxy-cafestol, 6,12-dihydroxy-cafestol, 2-oxo-cafestol, 6-oxo-cafestol and one isomer whose position in the carboxyl group was not determined. These metabolites were observed during 9 h of the experiment, whose contents were associated with the behavioral responses of the fish.