Search

Centella asiatica, or pegagan in Indonesia, is a perennial plant used in Indonesian traditional medicine (jamu) and functional food with many biological activities. Those biological activities come from the bioactive metabolites present in C. asiatica. Differences in metabolite pathways in each part of the plant affect the accumulation of metabolites contained, thus impacting its biological activity. Therefore, this study aims to identify and evaluate differences in the distribution of metabolites in each part of C. asiatica, namely leaves, stems, stolons, and roots. Each plant part was extracted using methanol and sonicated for 30 minutes. The metabolites in the samples were separated and detected using UHPLC-QOrbitrap HRMS. Differences in the distribution of metabolites in each part of the plant were evaluated using chemometrics analysis. UHPLC-Q-Orbitrap HRMS analysis could positively identify 37 metabolites, most of which belong to the phenylpropanoid, triterpenoid, triterpenoid saponin, and flavonoid groups. Principal component analysis was able to clearly distinguish each part of the plant using the peak intensity of the overall chromatogram and the peak area of the identified metabolites. The different biosynthetic pathways of metabolites in plants could cause a difference in the distribution of metabolites in each plant. [ABSTRACT FROM AUTHOR]

ABSTRACT: Complete self-assembly and reassembly behavior of bitter peptide-protein necessitates multilevel theories that encompass phenomena ranging from the self-assembly of recombinant complex to atomic trajectories. An extension to the level of mechanism method was put forth, involves limited enzymatic digestion and bottom-up proteomics to dissect inherent heterogeneity within β-LG and β-LG-PPGLPDKY complex and uncover conformational and dynamic alterations occurring in specific local regions of the model protein. Bitter peptide PPGLPDKY spontaneously bound to IIAEKTK, IDALNENK, and YLLFCMENSAEPEQSLACQCLVR regions of β-LG in a 1:1 stoichiometric ratio to mask bitterness perception. Molecular dynamic simulation and free energy calculation provided time-varying atomic trajectories of the recombinant complex and found that a peptide was stabilized in the upper region of the hydrophobic cavity with the binding free energy of −30.56 kJ mol−1 through 4 hydrogen bonds (Glu74, Glu55, Lys69, and Ser116) and hydrophobic interactions (Asn88, Asn90, and Glu112). Current research aims to provide valuable physical insights into the macroscopic self-assembly behavior between proteins and bitter peptides, and the meticulous design of highly acceptable taste characteristics in goat milk products.

Complete self-assembly and reassembly behavior of bitter peptide-protein necessitates multilevel theories that encompass phenomena ranging from the self-assembly of recombinant complex to atomic trajectories. An extension to the level of mechanism method was put forth, involves limited enzymatic digestion and bottom-up proteomics to dissect inherent heterogeneity within β-LG and β-LG-PPGLPDKY complex and uncover conformational and dynamic alterations occurring in specific local regions of the model protein. Bitter peptide PPGLPDKY spontaneously bound to IIAEKTK, IDALNENK, and YLLFCMENSAEPEQSLACQCLVR regions of β-LG in a 1:1 stoichiometric ratio to mask bitterness perception. Molecular dynamic simulation and free energy calculation provided time-varying atomic trajectories of the recombinant complex and found that a peptide was stabilized in the upper region of the hydrophobic cavity with the binding free energy of −30.56 kJ mol−1 through 4 hydrogen bonds (Glu74, Glu55, Lys69, and Ser116) and hydrophobic interactions (Asn88, Asn90, and Glu112). Current research aims to provide valuable physical insights into the macroscopic self-assembly behavior between proteins and bitter peptides, and the meticulous design of highly acceptable taste characteristics in goat milk products. [ABSTRACT FROM AUTHOR]

Epimedium-Rhizoma drynariae (EP-RD) was a well-known herb commonly used to treat bone diseases in traditional Chinese medicine. Nevertheless, there was incomplete pharmacokinetic behavior, metabolic conversion and chemical characterization of EP-RD in vivo. Therefore, this study aimed to establish metabolic profiles combined with multicomponent pharmacokinetics to reveal the in vivo behavior of EP-RD. Firstly, the diagnostic product ions (DPIs) and neutral losses (NLs) filtering strategy combined with UHPLC-Q-Orbitrap HRMS for the in vitro chemical composition of EP-RD and metabolic profiles of plasma, urine, and feces after oral administration of EP-RD to rats were proposed to comprehensively characterize the 47 chemical compounds and the 97 exogenous in vivo (35 prototypes and 62 metabolites), and possible biotransformation pathways of EP-RD were proposed, which included phase I reactions such as hydrolysis, hydrogenation, dehydrogenation, hydroxylation, dehydroxylation, isomerization, and demethylation and phase II reactions such as glucuronidation, acetylation, methylation, and sulfation. Moreover, a UHPLC-MS/MS quantitative approach was established for the pharmacokinetic analysis of seven active components: magnoflorine, epimedin A, epimedin B, epimedin C, icariin, baohuoside II, and icariin II. Results indicated that the established method was reliably used for the quantitative study of plasma active ingredients after oral administration of EP-RD in rats. Compared to oral EP alone, the increase in area under curves and maximum plasma drug concentration (P < 0.05). This study increased the understanding of the material basis and biotransformation profiles of EP-RD in vivo, which was of great significance in exploring the pharmacological effects of EP-RD. [ABSTRACT FROM AUTHOR]

The present study investigates the chemical composition variances among Pinelliae Rhizoma, a widely used Chinese herbal medicine, and its common adulterants including Typhonium flagelliforme, Arisaema erubescens, and Pinellia pedatisecta. Utilizing the non-targeted metabolomics technique of employing UHPLC-Q-Orbitrap HRMS, this research aims to comprehensively delineate the metabolic profiles of Pinelliae Rhizoma and its adulterants. Multivariate statistical methods including PCA and OPLS-DA are employed for the identification of differential metabolites. Volcano plot analysis is utilized to discern upregulated and downregulated compounds. KEGG pathway analysis is conducted to elucidate the differences in metabolic pathways associated with these compounds, and significant pathway enrichment analysis is performed. A total of 769 compounds are identified through metabolomics analysis, with alkaloids being predominant, followed by lipids and lipid molecules. Significant differential metabolites were screened out based on VIP > 1 and p-value < 0.05 criteria, followed by KEGG enrichment analysis of these differential metabolites. Differential metabolites between Pinelliae Rhizoma and Typhonium flagelliforme, as well as between Pinelliae Rhizoma and Pinellia pedatisecta, are significantly enriched in the biosynthesis of amino acids and protein digestion and absorption pathways. Differential metabolites between Pinelliae Rhizoma and Arisaema erubescens are mainly enriched in tyrosine metabolism and phenylalanine metabolism pathways. These findings aim to provide valuable data support and theoretical references for further research on the pharmacological substances, resource development and utilization, and quality control of Pinelliae Rhizoma. [ABSTRACT FROM AUTHOR]

Copyright of Farmacia is the property of Societatea de Stiinte Farmaceutice Romania and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: To compare the effect of fermentation on the chemical constituents of Gastrodia Tuder Halimasch Powder (GTHP), to establish its fingerprinting and multicomponent content determination, and to provide a basis for the processing, handling, and clinical application of this herb. Methods: Ultra-high-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to conduct a preliminary analysis of the chemical constituents in GTHP before and after fermentation. High-performance liquid chromatography (HPLC) was used to determine some major differential components of GTHP and establish fingerprints. Cluster analysis (CA), and principal component analysis (PCA) were employed for comprehensive evaluation. Results: Seventy-nine compounds were identified, including flavonoids, organic acids, nucleosides, terpenoids, and others. The CA and PCA results showed that ten samples were divided into three groups. Through standard control and HPLC analysis, 10 compounds were identified from 22 peaks, namely uracil, guanosine, adenosine, 5-hydroxymethylfurfural (5-HMF), daidzin, genistin, glycitein, daidzein, genistein, and ergosterol. After fermentation, GTHP exhibited significantly higher contents of uracil, guanosine, adenosine, 5-hydroxymethylfurfural, and ergosterol and significantly lower genistein and daidzein contents. Conclusions: The UHPLC-Q-Orbitrap HRMS and HPLC methods can effectively identify a variety of chemical components before and after the fermentation of GTHP. This study provides a valuable reference for further research on the rational clinical application and quality control improvement of GTHP. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Shenyang Pharmaceutical University is the property of Shenyang Pharmaceutical University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The present study investigates the chemical composition variances among Pinelliae Rhizoma, a widely used Chinese herbal medicine, and its common adulterants including Typhonium flagelliforme, Arisaema erubescens, and Pinellia pedatisecta. Utilizing the non-targeted metabolomics technique of employing UHPLC-Q-Orbitrap HRMS, this research aims to comprehensively delineate the metabolic profiles of Pinelliae Rhizoma and its adulterants. Multivariate statistical methods including PCA and OPLS-DA are employed for the identification of differential metabolites. Volcano plot analysis is utilized to discern upregulated and downregulated compounds. KEGG pathway analysis is conducted to elucidate the differences in metabolic pathways associated with these compounds, and significant pathway enrichment analysis is performed. A total of 769 compounds are identified through metabolomics analysis, with alkaloids being predominant, followed by lipids and lipid molecules. Significant differential metabolites were screened out based on VIP > 1 and p-value < 0.05 criteria, followed by KEGG enrichment analysis of these differential metabolites. Differential metabolites between Pinelliae Rhizoma and Typhonium flagelliforme, as well as between Pinelliae Rhizoma and Pinellia pedatisecta, are significantly enriched in the biosynthesis of amino acids and protein digestion and absorption pathways. Differential metabolites between Pinelliae Rhizoma and Arisaema erubescens are mainly enriched in tyrosine metabolism and phenylalanine metabolism pathways. These findings aim to provide valuable data support and theoretical references for further research on the pharmacological substances, resource development and utilization, and quality control of Pinelliae Rhizoma.

Objective: To compare the effect of fermentation on the chemical constituents of Gastrodia Tuder Halimasch Powder (GTHP), to establish its fingerprinting and multicomponent content determination, and to provide a basis for the processing, handling, and clinical application of this herb. Methods: Ultra-high-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to conduct a preliminary analysis of the chemical constituents in GTHP before and after fermentation. High-performance liquid chromatography (HPLC) was used to determine some major differential components of GTHP and establish fingerprints. Cluster analysis (CA), and principal component analysis (PCA) were employed for comprehensive evaluation. Results: Seventy-nine compounds were identified, including flavonoids, organic acids, nucleosides, terpenoids, and others. The CA and PCA results showed that ten samples were divided into three groups. Through standard control and HPLC analysis, 10 compounds were identified from 22 peaks, namely uracil, guanosine, adenosine, 5-hydroxymethylfurfural (5-HMF), daidzin, genistin, glycitein, daidzein, genistein, and ergosterol. After fermentation, GTHP exhibited significantly higher contents of uracil, guanosine, adenosine, 5-hydroxymethylfurfural, and ergosterol and significantly lower genistein and daidzein contents. Conclusions: The UHPLC-Q-Orbitrap HRMS and HPLC methods can effectively identify a variety of chemical components before and after the fermentation of GTHP. This study provides a valuable reference for further research on the rational clinical application and quality control improvement of GTHP.

Sida rhombifolia, also known as sidaguri in Indonesia, is a medicinal plant commonly used as a herbal medicine because of its metabolite and biological activities. One of the several factors that affect plant metabolite composition and concentration is the use of plant parts. In this study, the experiment aimed to identify the metabolite profile in the leaves and stem extracts of S. rhombifolia using UHPLC-Q-Orbitrap HRMS-based untargeted metabolomics. The samples were distinguished by principal component analysis (PCA). Extraction of metabolites was conducted by sonication for approximately 30 min with 70% ethanol as the extraction solvent; 28 metabolites were identified. Seven metabolites were identified only in the leaves, three were identified only in the stems, and 18 other metabolites were identified in both the leaves and stems. These metabolites were categorized as flavonoids, triterpenoids, alkaloids, coumarins, phenolic aldehydes, phenolic acids, ecdysteroids, fatty acids, and monoterpene lactones. Based on the classification results, PCA grouped the leaves and stem extracts of S. rhombifolia using the peak area variables of the identified metabolites. [ABSTRACT FROM AUTHOR]

Natural bear bile powder (NBBP), a traditional Chinese medicine, has been clinically used to treat liver dysfunction, cardiovascular and cerebrovascular diseases. Recently it has been proved that NBBP has a good effect on regulating lipid disorders. However, the active ingredients and the anti‐hyperlipidemic effects are unclear. The high‐fat diet (HFD) induced hyperlipidemic model rats were treated with NBBP for 4 weeks and were examined for, lipid profile, liver function parameters, hepatic antioxidant status and histopathology of liver, epididymal adipose tissues. The NBBP treatment effectively improved serum lipid levels and normalized antioxidant status. A total of 16 constituents of NBBP were charactered via UHPLC‐Q‐TOF‐HRMS. We screened eight potentially active compounds and corresponded to 75 anti‐hyperlipidemic target proteins. PPI analysis showed that ALB, TNF, EGFE, PPARG, and PPARA are core proteins. Pathway enrichment analysis showed that NBBP might treat hyperlipidemia (HLP) by regulating a series of signaling pathways involving immune system, inflammatory response, and metabolism. Molecular docking revealed that gene had high affinities to NBBP compounds. In conclusion, UHPLC‐Q‐TOF‐HRMS analysis, network pharmacology, and in vivo experiments in this study provided essential references for revealing the potential mechanism of NBBP in the prevention and treatment of HLP. [ABSTRACT FROM AUTHOR]

Therapeutic drug monitoring is critical to decrease the incidence rate of bleeding and thrombosis for personalized treatment with rivaroxaban, especially for drug interaction treatment, patients with renal dysfunction, elderly patients, patients with cardiovascular problems, and so on. In addition, an accurate analytical method is necessary for therapeutic drug monitoring. This study developed a ultra‐HPLC–tandem Orbitrap high‐resolution MS (UHPLC–Q‐Orbitrap HRMS) method to accurately identify and quantify rivaroxaban in rat plasma. The isotope internal standard method was applied for accurate quantification. Rivaroxaban‐d4 was selected as the isotope internal standard substance. The m/z 436.07263 ([M + H]+) was selected as the precursor ion and m/z 144.95085 and m/z 231.11259 were selected as the main product ions for rivaroxaban. The lower limit of quantification of rivaroxaban in plasma was 0.01 mg/L. The intra‐ and inter‐day precisions were ≤3.65% and ≤8.16%, while the recoveries ranged from 87.4% to 95.2%. This analysis method was simple, low cost, and easy to operate. The developed and validated method was subsequently applied to successfully investigate the pharmacokinetic parameters of rivaroxaban in rats after its oral administration. These results could be helpful to promote further research regarding the mechanisms of rivaroxaban and drug interaction, which can avoid false positives due to high‐precision identification of the proposed method. [ABSTRACT FROM AUTHOR]

Dehydrocostus lactone (DL) is among the representative ingredients of traditional Chinese medicine (TCM), with excellent anticancer, antibacterial, and anti-inflammatory activities. In this study, an advanced strategy based on ultra-high-performance liquid chromatography–quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC–Q-Orbitrap HRMS) was integrated to comprehensively explore the metabolic fate of DL in rats. First, prior to data collection, all biological samples (plasma, urine, and feces) were concentrated and purified using solid-phase extraction (SPE) pre-treatment technology. Then, during data collection, in the full-scan (FS) data-dependent acquisition mode, FS-ddMS2 was intelligently combined with FS-parent ion list (PIL)-dynamic exclusion (DE) means for targeted monitoring and deeper capture of more low-abundance ions of interest. After data acquisition, data-mining techniques such as high-resolution extracted ion chromatograms (HREICs), multiple mass defect filters (MMDFs), diagnostic product ions (DPIs), and neutral loss fragments (NLFs) were incorporated to extensively screen and profile all the metabolites in multiple dimensions. As a result, a total of 71 metabolites of DL (parent drug included) were positively or tentatively identified. The results suggested that DL in vivo mainly underwent hydration, hydroxylation, dihydrodiolation, sulfonation, methylation, dehydrogenation, dehydration, N-acetylcysteine conjugation, cysteine conjugation, glutathione conjugation, glycine conjugation, taurine conjugation, etc. With these inferences, we successfully mapped the "stepwise radiation" metabolic network of DL in rats, where several drug metabolism clusters (DMCs) were discovered. In conclusion, not only did we provide a refined strategy for inhibiting matrix effects and fully screening major-to-trace metabolites, but also give substantial data reference for mechanism investigation, in vivo distribution visualization, and safety evaluation of DL. [ABSTRACT FROM AUTHOR]

Microgreens represent a new generation of food products, commonly used to garnish and embellish culinary dishes, and recently associated with an increasing interest in their nutraceutical and phytochemical profiles. Four Apiaceae species: Pimpinella anisum L. (anise), Anthriscus cerefolium L. (chervil), Carum carvi L. (caraway), and Anethum graveolens L. (dill) were assessed for fresh yield, macro- and microminerals, total chlorophylls, total ascorbic acid, carotenoids, polyphenols, and their antioxidant activity. Anise was the species yielding the most (2.53 kg m−2) and having the highest lutein content (18.4 µg g−1 dry weight (DW)). Chervil and dill were characterized by the highest total ascorbic acid content (~151 mg AA g−1 fresh weight (FW)). The phenolic profile highlighted the presence of five flavonoid derivatives and 12 phenolic acid derivatives, with quinic acid derivatives being the most abundant phenols in the species tested. In addition, anise, caraway, and dill proved to be considerably rich in total polyphenols (~11056 μg g−1 DW). Caraway and dill were characterized by the highest antioxidant activity measured by the DPPH and ABTS methods, whereas the FRAP method revealed caraway as having the highest antioxidant activity. Such results highlight the potential of Apiaceae species as an alternative to other families which are commonly used for microgreens production. [ABSTRACT FROM AUTHOR]

Pueraria is a rich source of bioactive compounds, but there is a lack of comprehensive information concerning its composition. Therefore, a UHPLC-Q-Orbitrap HRMS method was developed to identify and quantify bioactive compounds in pueraria. Twelve isoflavones were quantified, with puerarin being the most abundant, followed by puerarin 6″-O-xyloside, 3′-methoxy puerarin, and 3′-hydroxy puerarin. A further 88 bioactive components in eight categories were also tentatively identified. The 12 isoflavones, except for genistein, exhibited α-glucosidase inhibitory activity. The binding of these compounds to the active site of α-glucosidase was confirmed via molecular docking analysis. These findings provide a basis for identifying pueraria as a promising functional food ingredient. [ABSTRACT FROM AUTHOR]

ABSTRACT: Infant milk formulas are designed to substitute human milk when breastfeeding is unavailable. In addition to human milk and milk-derived products, these formulas can be a vehicle of contaminants. In this work, a multiclass method based on the QuEChERS (quick, easy, cheap, effective, rugged, and safe) approach was developed for the simultaneous determination of contaminants (n = 45), including mycotoxins and veterinary drug residues, occurring in infant milk formulas. By using an ultra-high-performance liquid chromatography quadrupole-Orbitrap coupled with high-resolution mass spectrometry analysis (UHPLC-Q-Orbitrap HRMS; Thermo Fisher Scientific), further retrospective analysis of 337 contaminants, including pesticides, was achieved. The method was validated in accordance with European regulations and applied for the analysis of 54 infant milk samples. Risk assessment was also performed. Dexamethasone was detected in 16.6% of samples (range: 0.905–1.131 ng/mL), and procaine benzyl penicillin in 1 sample at a concentration of 0.295 ng/mL. Zearalenone was found in 55.5% of samples (range: 0.133–0.638 ng/mL) and α-zearalenol in 16.6% of samples (range: 1.534–10.408 ng/mL). Up to 49 pesticides, 11 veterinary drug residues, and 5 mycotoxins were tentatively identified via retrospective analysis based on the mass spectral library. These findings highlight the necessity of careful evaluation of contaminants in infant formulas, considering that they are intended for a vulnerable part of the population.

Objective: UPLC-Q-Orbitrap HRMS, network pharmacology and molecular docking technology were used to explore the material basis and mechanism of Schisandrae chinensis (SC) in treating non-alcoholic fatty liver disease(NAFLD). Method: UPLC-Q-Orbitrap HRMS was used to identify the chemical components in SC. The Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) and Swiss Target Prediction online platform were used to screen and predict the potential targets of the chemical composition of SC; GeneCards, CTD, OMIM, Dis-GeNET, GEO database were used to collect non-alcoholic fatty liver disease potential targets. String database and Cytoscape 3.7.1 software were used to construct a protein-protein interaction (PPI) network model; R-based bioconductor data package for gene ontology (GO) and KEGG pathway were used to analyse potential targets; Cytoscape 3.7.1 software were used to establish a "drug-key active ingredient-target-pathway" network. Finally, molecular docking was carried out to preliminarily verify the mechanism of SC in the treatment of NAFLD. Results: 50 components of SC were analyzed and identified, and 246 potential targets of SC in treating NAFLD were screened out. Through further analysis of network topology, 23 core components and 30 potential core targets were screened out, and enrichment analysis was carried out based on them. SC played a therapeutic role through the proteoglycans in cancer, endocrine resistance, Rap1 signaling pathway, VEGF signaling pathway, AGE-RAGE signaling pathway in diabetic complications and Estrogen signaling pathway. Molecular bonding results showed that the top 5 active ingredients, quercetin, kaempferol, schisandrin a, α-linolenic acid and schisandrol B, had good binding activities with the top 4 core targets, AKT1, HSP90AA1, SRC and MAPK1, and their binding free energies were all less than −5 kcal/mol. The docking conformation of the molecule was stable. Conclusion: SC may improve the symptoms of NAFLD by acting on lipid metabolism, oxidative stress, angiogenesis and inflammation related pathways and targets.

Ebony (Diospyros celebica Bakh) is an endemic forest tree species found in Sulawesi whose basic information on its metabolite profile is still lacking. The objective of this research was to separate and identify putatively metabolite present in the leaves and wood of ebony across Sulawesi. Separation and identification of ebony metabolites were carried out using UHPLC-Q-Orbitrap HRMS analysis. Using ultrasonication with ethanol as the extracting solvent, we used powdered ebony leaves and wood. The results showed that the metabolites contained in the leaves and wood of ebony were 59 metabolites. About 14 compounds were found in the leaves and wood, 21 compounds in the wood, and 24 compounds in the leaves. The identified metabolites are flavonoids, terpenoids, amino aldehydes, alkaloids, quinones, steroids, amino acids, fatty acids, and saccharides. Clustering of ebony using principal component analysis obtained leaves and wood groups using peak area of known compounds as the variable.

Gleditsiae Sinensis Fructus (GSF) is commonly used in traditional medicine to treat respiratory diseases such as bronchial asthma. However, there is a lack of research on the chemical composition of GSF and the pharmacological substance and mechanism of action for GSF in treating bronchial asthma. The chemical constituents of GSF were analyzed using ultrahigh-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). In this study, we combined network pharmacology, molecular docking techniques, and experimental validation to explore the therapeutic efficacy and underlying mechanism of GSF in the treatment of bronchial asthma. Characterization of the chemical constituents of GSF was conducted using UHPLC-Q-Orbitrap HRMS. The identified chemical components were subjected to screening for active ingredients in the Swiss Absorption, Distribution, Metabolism, and Excretion (ADME) database. Relevant databases were utilized to retrieve target proteins for the active ingredients and targets associated with bronchial asthma disease, and the common targets between the two were selected. Subsequently, the protein-protein interaction (PPI) network was constructed using the String database and Cytoscape software to identify key targets. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the Metascape database. The "component-common target" network was constructed using Cytoscape to identify the primary active ingredients. Molecular docking validation was conducted using AutoDock software. The bronchial asthma mouse model was established using ovalbumin (OVA), and the lung organ index of the mice was measured. Lung tissue pathological changes were observed using hematoxylin and eosin (HE), Periodic Acid-Schiff (PAS), and Masson staining. The respiratory resistance (Penh) of the mice was assessed using a pulmonary function test instrument. An enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of IgE, IL-4, IL-5, and IL-13 in the mouse serum. Immunofluorescence staining was performed to detect the protein expression levels of AKT and PI3K in the lung tissues. An in vitro experiment was performed to observe the effects of echinocystic acid (EA) on IL-4 stimulated Human ASMCs (hASMCs). Cell viability was measured using a CCK-8 assay to calculate the IC50 value of the EA. A wound healing test was conducted to observe the effect of EA on degree of healing. RT-qPCR was performed to detect the influence of EA on the mRNA expression levels of ALB, SRC, TNF-α, AKT1, and IL6 in the cells. A total of 95 chemical constituents were identified from the GSF. Of these, 37 were identified as active ingredients. There were 169 overlapping targets between the active ingredients and the disease targets. A topological analysis of the protein-protein interaction (PPI) network identified the core targets as IL6, TNF, ALB, AKT1, and SRC. An enrichment analysis revealed that the treatment of bronchial asthma with GSF primarily involved the AGE-RAGE signaling pathway and the PI3K-Akt signaling pathway, among others. The primary active ingredients included 13(s)-HOTRE, linolenic acid, and acacetin. The molecular docking results demonstrated a favorable binding activity between the critical components of GSF and the core targets. Animal experimental studies indicated that GSF effectively improved symptoms, lung function, and lung tissue pathological changes in the OVA-induced asthmatic mice, while alleviating inflammatory responses. GSF decreased the fluorescent intensity of the AKT and PI3K proteins. The IC50 value of EA was 30.02μg/ml. EA (30) significantly promoted the proliferation of IL4-stimulated hASMCs cells. EA (30) significantly increased the expression of ALB and SRC mRNA and decreased the expressions of TNF-α, AKT, and IL6 mRNA. The multiple active ingredients found in GSF exerted their anti-inflammatory effects through multiple targets and pathways. This preliminary study revealed the core target and the mechanism of action underlying its treatment of bronchial asthma. These findings provided valuable insights for further research on the pharmacological substances and quality control of GSF. [Display omitted] [ABSTRACT FROM AUTHOR]

Chuanhuang Qingre capsule (CHQR) is composed of Andrographis paniculata and solidago virgaurea, mainly used for treating acute respiratory infection, acute tonsillitis and acute pharyngitis. However, the mechanism of CHQR in treating pharyngitis is still unclear. In this study, UHPLC‐Q‐Orbitrap HRMS technology was used to identify the chemical constituents in CHQR. Network pharmacology combined with a molecular docking approach was used to screen for possible mechanisms to predict CHQR for treating acute pharyngitis. Finally, a rat model of acute pharyngitis was used to confirm the mechanism of CHQR in treating acute pharyngitis.The results showed that UHPLC‐Q‐Orbitrap HRMS identified 68 compounds. Network analysis showed that TP53, AKT1, TNF, IL6 and EGFR were the key targets. The results of molecular docking showed reasonable binding force. The experimental results showed that CHQR could reduce inflammatory cell infiltration and improve pharyngitis symptoms in rats with acute pharyngitis. Elisa results showed that the levels of PGE2, IL‐1β, TNF‐α and IL‐6 in serum decreased. It is suggested that CHQR can prevent and treat acute pharyngitis, mainly by inhibiting the TNF pathway and reducing of inflammatory cytokines levels, which can provide scientific references for clinical research. [ABSTRACT FROM AUTHOR]

• Total 75 diverse chemical constituents were identified from B. gymnorhiza leaves. • LC-MS analysis identified 60 compounds belonging to various classes, including flavonoids, fatty acids, terpenes, carboxylic acids, phenolic acids, quinones, and alkaloids. • GC–MS analysis revealed presence of more saturated fatty acid content. This study examines the chemical compositions of most common and widely distributed mangrove species Bruguiera gymnorhiza. Here, the metabolic profiling of B. gymnorhiza leaves via gas chromatography-mass spectrometry and an advanced ultra-high performance liquid chromatography-Q-Exactive Orbitrap high resolution mass spectrometry was performed to screen and identify the bioactive chemical constituents. Total of 75 diverse chemical constituents were identified which belong to various classes, including flavonoids, fatty acids, terpenoids, carboxylic acids, phenolic acids, quinones, and alkaloid. Among these metabolites flavonoids, fatty acids and terpenoids were found to be the major component of B. gymnorhiza. Moreover, the compounds which are key drivers of pharmacological actions of plants such as, gallic acid, hispidulin, rhamnetin, embelin and 5-O-methyl embelin were reported. This study is the first report on the HRMS analysis of chemical constituent of B. gymnorhiza and can be considered as comprehensive map of its biologically important metabolites. [ABSTRACT FROM AUTHOR]

The leaves of Annona muricata (sirsak), Gynura procumbens (sambung nyawa), and Typhonium flagelliforme (keladi tikus) have been used as traditional medicines in Indonesia. This study aims to determine the antioxidant capacity and putatively identified phenolics from the leaves of three medicinal plants forementioned above. We used the DPPH (2,2-diphenyl-1-picrylhydrazyl) method for measuring radical scavenging (antioxidant assay) while the phenolics profiling was determined using UHPLC-Q-Orbitrap HRMS. The results showed that the percentage of radical scavenging activity of G. procumbens leaves extract in ethanol was higher than the other two plants. Phenolics profiling of the three medicinal plants was identified with 38 compounds belonging to flavones and flavanols hydroxycinnamic acid, and several other groups. The number of metabolites identified putatively was 12, 31, and 19 metabolites in the extracts of A. muricata, G. procumbens, and T. flagelliforme, respectively. The results confirmed the correlation between the phenolics presence and the antioxidant capacity of three plants used in this study.

Copyright of Journal of Chinese Institute of Food Science & Technology is the property of Journal of Chinese Institute of Food Science & Technology Periodical Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Polygonum capitatum as an ethnic medicine has been used to treat urinary tract infections, pyelonephritis and urinary calculi. In our previous study, P. capitatum was found to have anti-hyperuricemia effects. Nevertheless, the active constituents of P. capitatum for treating hyperuricemia were still unclear. In this study, an ultra-high-performance liquid chromatography coupled to quadrupole/orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to comprehensively detect the chemical ingredients of P. capitatum and its absorbed constituents in the plasma of hyperuricemia rats for the first time. Xcalibur 3.0 and Compound Discoverer 2.0 software coupled to mzCloud and ChemSpider databases were utilized for qualitative analysis. A total of 114 chemical components including phenolics, flavonoids, tannins, phenylpropanoids, amino acids, amides and others were identified or tentatively characterized based on the exact mass, retention time and structural information. Compared to the previous P. capitatum study, an additional 66 different components were detected. Moreover, 68 related xenobiotics including 16 prototype components and 52 metabolites were found in the plasma of hyperuricemia rats. The metabolic pathways included ring fission, hydrolysis, decarboxylation, dehydroxylation, methylation, glucuronidation and sulfation. This work may provide important information for further investigation on the active constituents of P. capitatum and their action mechanisms for anti-hyperuricemia effects. [ABSTRACT FROM AUTHOR]

Ranunculus sceleratus L.(RS) has shown various pharmacological effects in traditional Chinese medicine. In our previous study, the positive therapeutic effect on α-naphthylisothiocyanate induced intrahepatic cholestasis in rats was obtained using TianJiu treatment with fresh RS. However, the chemical profile of RS has not been clearly clarified, which impedes the research progress on the therapeutic effect of RS. Herein, an ultra-high performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) method was developed to rapidly separate and identify multiple constituents in the 80% methanol extract of RS. A total of sixty-nine compounds (19 flavonoids, 22 organic acids, 6 coumarins, 4 lignans, 14 nitrogenous compounds, and 4 anthraquinones) were successfully characterized. A total of 12 of these compounds were unambiguously identified by standard samples. Their mass spectrometric fragmentation pathways were investigated. It is worth noting that flavonoids and lignans were identified for the first time in RS. In this study, we successfully provide the first comprehensive report on identifying major chemical constituents in RS by UHPLC-Q-Orbitrap HRMS. The obtained results enrich the RS chemical profile, paving the way for further phytochemical study, quality control, and pharmacological investigation of RS. [ABSTRACT FROM AUTHOR]

Organophosphate esters (tri-OPEs), which are a significant group of emerging organic pollutants, have been widely found in aquatic products worldwide. While tri-OPEs have been extensively detected in aquatic products and are known to represent potential risks to human health, there is limited information available regarding their metabolites (di-OPEs). Currently, the techniques for analyzing the composition of tri- and di-OPEs have been distinguished and have certain drawbacks in terms of their expense, time requirements, and reliability. The objective of the study is to develop, validate, and utilize a modified QuEChERS technique for extracting tri- and di-OPEs in marine fish. As a result, EMR-Lipid and WAX were selected as adsorbents for the QuEChERS method to purify the samples and a combination of centrifugal vacuum and nitrogen at a temperature of 1 °C was selected as a technique to enhance the sensitivity of the developed method. The parameters of the method validation are in accordance with the criteria specified in SANTE/11312/2021 (a recovery ranging from 84.0% to 107.4% and RSD of precision below 20%). The aforementioned methodology was employed to assess the tri-OPEs and their metabolites in a total of 72 fish muscle samples obtained from the presence of OPEs in all recorded instances. The average concentration of ΣOPEs was determined to be 87.4±55.0 ng/g dw, with TBOEP and BEHP identified as the predominant compounds. Furthermore, no significant variation in the content of tri- and di-OPEs was observed in fish samples, regardless of species or geographical location. [Display omitted] • The modified QuEChERS extracts tri- and di-OPEs from lipid-rich fish matrices. • UHPLC-HRMS was used to measure tri- and di-OPEs in the fish samples. • Good quantification with high recovery and repeatability was achieved. • Vietnamese marine fish have strong di- and tri-OPE connections. [ABSTRACT FROM AUTHOR]

Infant milk formulas are designed to substitute human milk when breastfeeding is unavailable. In addition to human milk and milk-derived products, these formulas can be a vehicle of contaminants. In this work, a multiclass method based on the QuEChERS (quick, easy, cheap, effective, rugged, and safe) approach was developed for the simultaneous determination of contaminants (n = 45), including mycotoxins and veterinary drug residues, occurring in infant milk formulas. By using an ultra-high-performance liquid chromatography quadrupole-Orbitrap coupled with high-resolution mass spectrometry analysis (UHPLC-Q-Orbitrap HRMS; Thermo Fisher Scientific), further retrospective analysis of 337 contaminants, including pesticides, was achieved. The method was validated in accordance with European regulations and applied for the analysis of 54 infant milk samples. Risk assessment was also performed. Dexamethasone was detected in 16.6% of samples (range: 0.905–1.131 ng/mL), and procaine benzyl penicillin in 1 sample at a concentration of 0.295 ng/mL. Zearalenone was found in 55.5% of samples (range: 0.133–0.638 ng/mL) and α-zearalenol in 16.6% of samples (range: 1.534–10.408 ng/mL). Up to 49 pesticides, 11 veterinary drug residues, and 5 mycotoxins were tentatively identified via retrospective analysis based on the mass spectral library. These findings highlight the necessity of careful evaluation of contaminants in infant formulas, considering that they are intended for a vulnerable part of the population. [ABSTRACT FROM AUTHOR]

The dessert bananas genotype are typically AA, AAA, and AAB, while ABB is classified as cooking bananas. Their nutritional content can be accessed much better than cooking bananas by the human body because they can be consumed directly without processing. This present study aimed to investigate the metabolite profiles of the ripe pulp of ten Indonesian dessert bananas cultivars represents the commercial interest and unpopular cultivars groups by UHPLC-Q-Orbitrap HRMS. They were Ambon Putih, Ambon Kuning, Ambon Badak, Ambon Lumut, Raja Sereh, Raja Bulu, Lampung, Papan, Rejang, and Udang. Nineteen metabolites were putatively identified in all of the cultivars studied. The main groups were fatty acids and the others catecholamine, quinone, phenol, flavonoid, cinnamic acid, organic acid, indole, and amino acid groups. Some of them were important bioactive compounds such as linolenic acid, palmitelaidic acid, linoleic acid, p-coumaric acid, dopamine, myricetin-deoxyhexose-hexoside, eugenol, 1,4-benzoquinone, and antroquinonol. The PCA score and loading plot showed the ten cultivars were separated into two groups based on linolenic acid as the first (PC1, 57%) and p-coumaric acid (PC2, 38%) as the second principal component with the 95% total variance explained. There was no relationship between PCA grouping and the genome group in terms of composition and metabolite content. These results provide valuable information for plant breeding programs, especially in banana biofortification for the future. [ABSTRACT FROM AUTHOR]

Xiaofang Jiang,1,* Lin Zhou,2,3,* Lihua Zuo,2,3 Xiaohui Wang,4 Yingying Shi,2,3 Xiangyu Du,5 Jun Zhang,2,3 Liwei Liu,2,3 Zhuolun Li,2,3 Lianping Xue,2,3 Xin Liu,1 Zhi Sun2,3 1Department of Traditional Chinese Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province 450052, People’s Republic of China; 2Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province 450052, People’s Republic of China; 3Henan Key Laboratory of Precision Clinical Pharmacy, Zhengzhou, Henan Province 450052, People’s Republic of China; 4Department of Ultrasonography, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province 450052, People’s Republic of China; 5School of Basic Medical Science, Henan University, Kaifeng, Henan Province 475001, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhi Sun; Lin ZhouDepartment of Pharmacy, The First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Erqi District, Zhengzhou 450052, Henan Province, People’s Republic of ChinaTel/Fax +86 371 66862570Email sunzhi2013@163.com; fcczhoul@zzu.edu.cnPurpose: Shenkang injection, a traditional Chinese herbal prescription, had been widely used in renal disease due to its perfect curative effect. In this research, a novel, sensitive, accurate and rapid liquid chromatography-tandem mass spectrometric method was developed to simultaneously detect the seven active ingredients in rat plasma of Shenkang injection and investigate its pharmacokinetic behaviors with metabolism profiling meanwhile.Methods: For accurate pharmacokinetic quantitation, a WATERS ACQUITY UPLC® BEH C18 column was used to perform a separation and acetonitrile-water (0.1% formic acid) was selected as mobile phase for gradient elution with a flow rate of 0.20 mL/min. A heated electrospray ionization with selective reaction monitoring mode was used to monitor the precursor-product ion transitions for all the analytes and IS.Results: They all showed good linearity over a wide concentration range (r> 0.996 3) and the lower limit of quantification (LLOQ) was 0.1– 1.0 ng/mL for analytes. The validation parameters were all within the acceptable limits. Furthermore, for metabolism profiling study, metabolites of the seven ingredients were identified from the rat plasma based on the accurate mass and fragment ions. The metabolic pathways mainly focus on reduction, dehydration and conjugation.Conclusion: This study provided an overview of disposition of Shenkang injection, which is highly instructive for better understanding the effectiveness and toxicity of this drug.Keywords: Shenkang injection, pharmacokinetics, metabolism profiling, UHPLC-MS/MS, UHPLC-Q-orbitrap HRMS

Dehydrocostus lactone (DL) is among the representative ingredients of traditional Chinese medicine (TCM), with excellent anticancer, antibacterial, and anti-inflammatory activities. In this study, an advanced strategy based on ultra-high-performance liquid chromatography–quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC–Q-Orbitrap HRMS) was integrated to comprehensively explore the metabolic fate of DL in rats. First, prior to data collection, all biological samples (plasma, urine, and feces) were concentrated and purified using solid-phase extraction (SPE) pre-treatment technology. Then, during data collection, in the full-scan (FS) data-dependent acquisition mode, FS-ddMS2 was intelligently combined with FS-parent ion list (PIL)-dynamic exclusion (DE) means for targeted monitoring and deeper capture of more low-abundance ions of interest. After data acquisition, data-mining techniques such as high-resolution extracted ion chromatograms (HREICs), multiple mass defect filters (MMDFs), diagnostic product ions (DPIs), and neutral loss fragments (NLFs) were incorporated to extensively screen and profile all the metabolites in multiple dimensions. As a result, a total of 71 metabolites of DL (parent drug included) were positively or tentatively identified. The results suggested that DL in vivo mainly underwent hydration, hydroxylation, dihydrodiolation, sulfonation, methylation, dehydrogenation, dehydration, N-acetylcysteine conjugation, cysteine conjugation, glutathione conjugation, glycine conjugation, taurine conjugation, etc. With these inferences, we successfully mapped the “stepwise radiation” metabolic network of DL in rats, where several drug metabolism clusters (DMCs) were discovered. In conclusion, not only did we provide a refined strategy for inhibiting matrix effects and fully screening major-to-trace metabolites, but also give substantial data reference for mechanism investigation, in vivo distribution visualization, and safety evaluation of DL.

Microgreens represent a new generation of food products, commonly used to garnish and embellish culinary dishes, and recently associated with an increasing interest in their nutraceutical and phytochemical profiles. Four Apiaceae species: Pimpinella anisum L. (anise), Anthriscus cerefolium L. (chervil), Carum carvi L. (caraway), and Anethum graveolens L. (dill) were assessed for fresh yield, macro- and microminerals, total chlorophylls, total ascorbic acid, carotenoids, polyphenols, and their antioxidant activity. Anise was the species yielding the most (2.53 kg m−2) and having the highest lutein content (18.4 µg g−1 dry weight (DW)). Chervil and dill were characterized by the highest total ascorbic acid content (~151 mg AA g−1 fresh weight (FW)). The phenolic profile highlighted the presence of five flavonoid derivatives and 12 phenolic acid derivatives, with quinic acid derivatives being the most abundant phenols in the species tested. In addition, anise, caraway, and dill proved to be considerably rich in total polyphenols (~11056 μg g−1 DW). Caraway and dill were characterized by the highest antioxidant activity measured by the DPPH and ABTS methods, whereas the FRAP method revealed caraway as having the highest antioxidant activity. Such results highlight the potential of Apiaceae species as an alternative to other families which are commonly used for microgreens production.

Pueraria is a rich source of bioactive compounds, but there is a lack of comprehensive information concerning its composition. Therefore, a UHPLC-Q-Orbitrap HRMS method was developed to identify and quantify bioactive compounds in pueraria. Twelve isoflavones were quantified, with puerarin being the most abundant, followed by puerarin 6″-O-xyloside, 3′-methoxy puerarin, and 3′-hydroxy puerarin. A further 88 bioactive components in eight categories were also tentatively identified. The 12 isoflavones, except for genistein, exhibited α-glucosidase inhibitory activity. The binding of these compounds to the active site of α-glucosidase was confirmed via molecular docking analysis. These findings provide a basis for identifying pueraria as a promising functional food ingredient.

Polygonum hydropiper L. (PL) is widely used in treating enteritis in China. This study interpreted the active ingredient and mechanism of PL against enteritis through UHPLC‐Q‐Orbitrap HRMS, network pharmacology and molecular docking. UHPLC‐Q‐Orbitrap HRMS of PL manifested 68 compounds. In addition, the primary candidate genes and potential active components were identified by topological analysis of the single‐component disease gene interaction network. The interaction between the active ingredient, whose drug‐likeness properties were confirmed by Lipinski's rule, and the therapeutic gene was confirmed by molecular docking analysis. AutoDock Vina in AutoDock Tools was used to conduct molecular docking between significant components and critical genes. The advantage of this experiment is that the UHPLC‐Q‐Orbitrap HRMS method increases the total chemical composition of PL. Secondly, we obtained active compounds through network pharmacology and found that PL mainly acts on multiple inflammatory pathways in the treatment of enteritis. 61 potential genes of PL for the treatment of enteritis were obtained. The genes were mainly involved in biological processes such as response to oxidative stress, inhibition of inflammatory factors and inflammatory pathways, and role in treating enteritis by participating in AR, NOS2, MMP9, MMP2, SRC, and other signaling pathways. [ABSTRACT FROM AUTHOR]

Introduction: Individuals with metabolic syndrome (MetS) are at increasing risk of coronary artery disease (CAD). We investigated the common metabolic perturbations of CAD and MetS via serum metabolomics to provide insight into potential associations. Methods: Non-targeted serum metabolomics analyses were performed using ultra high-performance liquid chromatography coupled with Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry (UHPLC-Q-Orbitrap HRMS) in samples from 492 participants (272 CAD vs. 121 healthy controls (HCs) as cohort 1, 55 MetS vs. 44 HCs as cohort 2). Cross-sectional data were obtained when the participants were recruited from the First Affiliated Hospital of Zhengzhou University. Multivariate statistics and Student's t test were applied to obtain the significant metabolites [with variable importance in the projection (VIP) values >1.0 and p values <0.05] for CAD and MetS. Logistic regression was performed to investigate the association of identified metabolites with clinical cardiac risk factors, and the association of significant metabolic perturbations between CAD and MetS was visualized by Cytoscape software 3.6.1. Finally, the receiver operating characteristic (ROC) analysis was evaluated for the risk prediction values of common changed metabolites. Results: Thirty metabolites were identified for CAD, mainly including amino acids, lipid, fatty acids, pseudouridine, niacinamide; 26 metabolites were identified for MetS, mainly including amino acids, lipid, fatty acids, steroid hormone, and paraxanthine. The logistic regression results showed that all of the 30 metabolites for CAD, and 15 metabolites for MetS remained significant after adjustments of clinical risk factors. In the common metabolic signature association analysis between CAD and MetS, 11 serum metabolites were significant and common to CAD and MetS outcomes. Out of this, nine followed similar trends while two had differing directionalities. The nine common metabolites exhibiting same change trend improved risk prediction for CAD (86.4%) and MetS (90.9%) using the ROC analysis. Conclusion: Serum metabolomics analysis might provide a new insight into the potential mechanisms underlying the common metabolic perturbations of CAD and MetS. [ABSTRACT FROM AUTHOR]

IntroductionIndividuals with metabolic syndrome (MetS) are at increasing risk of coronary artery disease (CAD). We investigated the common metabolic perturbations of CAD and MetS via serum metabolomics to provide insight into potential associations.MethodsNon-targeted serum metabolomics analyses were performed using ultra high-performance liquid chromatography coupled with Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry (UHPLC-Q-Orbitrap HRMS) in samples from 492 participants (272 CAD vs. 121 healthy controls (HCs) as cohort 1, 55 MetS vs. 44 HCs as cohort 2). Cross-sectional data were obtained when the participants were recruited from the First Affiliated Hospital of Zhengzhou University. Multivariate statistics and Student’s t test were applied to obtain the significant metabolites [with variable importance in the projection (VIP) values >1.0 and p values

The current study aimed to explore the mechanism of Guizhi Shaoyao Zhimu Decoction in the treatment of gouty arthritis based on the combination of Vanquish Ultra High Performance Liquid Chromatography Q Exactive IV Pole-electrostatic field orbital trap high resolution mass spectrometer (UHPLC-Q-Orbitrap HRMS) and network pharmacology. Firstly, UHPLC-Q-Orbitrap HRMS was established and applied to separation and identification of the chemical components of GSZD. Then, using network pharmacology analysis to obtain the potential active components and candidate targets underlying the effect of GSZD on gouty arthritis (GA). Through targeted and untargeted analysis, a total of 79 components, including 3 chromones, 26 flavonoids, 10 alkaloids, 10 phenolic acids, 7 nucleosides and nucleobases, and 23 other components, were characterized from GSZD. 11 compounds were considered to potentially key active compounds against GA which may be via interacting with multiple inflammatory factors and multiple inflammatory pathways.

The incidence of cerebral ischemic stroke characterized by high mortality is increasing every year. Danshen Chuanxiongqin Injection (DSCXQ), a traditional Chinese medicine (TCM) preparation, is often applied to treat cerebral apoplexy and its related sequelae. However, there is a lack of systematic research on how DSCXQ mediates its protective effects against cerebral ischemia stroke. Metabolomic analysis based on UHPLC-Q-Orbitrap HRMS was employed to explore the potential mechanisms of DSCXQ on ischemic stroke induced by transient middle cerebral artery occlusion (MCAO). Pattern analysis and metabolomic profiling, combined by multivariate analysis disclosed that 55 differential metabolites were identified between Sham group and Model group, involving sphingolipid metabolism, glycerophospholipid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, primary bile acid biosynthesis, pantothenate and CoA synthesis and valine, leucine and isoleucine biosynthesis pathways. DSCXQ could reverse brain metabolic deviations in stroke by significantly upregulating the levels of L-tryptophan, Lyso (18:0/0:0), LPC (18:2), Indole-3-methyl acetate, and downregulating the levels of sphinganine 1-phosphate, L-threonic acid, glutaconic acid and N6,N6,N6-Trimethyl-L-lysine. In our study, we focused on the neuroprotective effects of DSCXQ against neuroinflammatory responses and neuronal apoptosis on a stroke model based on sphingolipid metabolism. The expressions of Sphk1, S1PR1, CD62P, Bcl-2, Bax, and cleaved Caspase-3 in brain tissue were evaluated. The neurological deficit, cerebral infarct size and behavioral abnormality were estimated. Results showed that DSCXQ intervention significantly reduced cerebral infarct size, ameliorated behavioral abnormality, inhibited the expression of Sphk1, S1PR1, CD62P, Bax, Cleaved Caspase-3, while increased the level of Bcl-2, and prevented neuronal apoptosis. The limitations are that our study mainly focused on the verification of sphingolipid metabolism pathway in stroke, and while other metabolic pathways left unverified. Our study indicates that SphK1-SIP axis may potentiate neuroinflammatory responses and mediate brain damage through neuronal apoptosis, and DSCXQ could suppress the activity of SphK1-SIP axis to protect brain tissue in cerebral ischemia. In conclusion, this study facilitates our understanding of metabolic changes in ischemia stroke and the underlying mechanisms related to the clinical application of DSCXQ.

Ranunculus sceleratus L.(RS) has shown various pharmacological effects in traditional Chinese medicine. In our previous study, the positive therapeutic effect on α-naphthylisothiocyanate induced intrahepatic cholestasis in rats was obtained using TianJiu treatment with fresh RS. However, the chemical profile of RS has not been clearly clarified, which impedes the research progress on the therapeutic effect of RS. Herein, an ultra-high performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) method was developed to rapidly separate and identify multiple constituents in the 80% methanol extract of RS. A total of sixty-nine compounds (19 flavonoids, 22 organic acids, 6 coumarins, 4 lignans, 14 nitrogenous compounds, and 4 anthraquinones) were successfully characterized. A total of 12 of these compounds were unambiguously identified by standard samples. Their mass spectrometric fragmentation pathways were investigated. It is worth noting that flavonoids and lignans were identified for the first time in RS. In this study, we successfully provide the first comprehensive report on identifying major chemical constituents in RS by UHPLC-Q-Orbitrap HRMS. The obtained results enrich the RS chemical profile, paving the way for further phytochemical study, quality control, and pharmacological investigation of RS.

Polygonum capitatum as an ethnic medicine has been used to treat urinary tract infections, pyelonephritis and urinary calculi. In our previous study, P. capitatum was found to have anti-hyperuricemia effects. Nevertheless, the active constituents of P. capitatum for treating hyperuricemia were still unclear. In this study, an ultra-high-performance liquid chromatography coupled to quadrupole/orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to comprehensively detect the chemical ingredients of P. capitatum and its absorbed constituents in the plasma of hyperuricemia rats for the first time. Xcalibur 3.0 and Compound Discoverer 2.0 software coupled to mzCloud and ChemSpider databases were utilized for qualitative analysis. A total of 114 chemical components including phenolics, flavonoids, tannins, phenylpropanoids, amino acids, amides and others were identified or tentatively characterized based on the exact mass, retention time and structural information. Compared to the previous P. capitatum study, an additional 66 different components were detected. Moreover, 68 related xenobiotics including 16 prototype components and 52 metabolites were found in the plasma of hyperuricemia rats. The metabolic pathways included ring fission, hydrolysis, decarboxylation, dehydroxylation, methylation, glucuronidation and sulfation. This work may provide important information for further investigation on the active constituents of P. capitatum and their action mechanisms for anti-hyperuricemia effects.

Kadsura heteroclita (Roxb) Craib stem(KHS) is amedicinal plant used for the treatment of rheumatismarthritis diseases in Tujia ethnomedicine. Thus far, the complex chemical compositions in KHS are not clear, and the levels of the major compounds in KHS are not well understood. In this study, a novel UHPLC-Q-Orbitrap HRMS method was established for the simultaneous quali-quantitative analysis of KHS. A total of 204 compounds were identified, including triterpenoids, lignans, sesquiterpenes, fatty acids, phenolic acids, and flavonoids, more than 100 of which were first discovered in KHS. Using the same method, 12 representative bioactive components were successfully quantified. The method was fully validated by linearity, LOD, LOQ, precision, stability, recovery, and matrix effects, and it was applied to quantify the 12 representative compounds in 4 batches ofKHS.As thismethod enables retrospective data analysis and has no upper limit to the number of analytes in a single run, it can be applied to quantify more active components of KHS in the future. [ABSTRACT FROM AUTHOR]

Background: Oral lichen planus (OLP) is a T-cell-mediated chronic inflammatory disorder and precancerous oral lesion with high incidence. The current diagnostic method of OLP is very limited and metabolomics may provide a new approach for quantitative evaluation.Methods: The Ultra-Performance Liquid Chromatography-Quadrupole/Orbitrap High Resolution Mass Spectrometry (UHPLC-Q-Orbitrap HRMS) was applied to analyze the change of metabolites in serum of patients with OLP. A total of 115 OLP patients and 124 healthy controls were assigned to either a training set (n = 160) or a test set (n = 79). The potential biomarkers and the change of serum metabolites were profiled and evaluated by multivariate analysis.Results: Totally, 23 differential metabolites were identified in the training set between OLP group and healthy group. Three prominent metabolites in receiver operating characteristic (ROC) were selected as a panel to distinguish OLP or healthy individuals in the test set, and the diagnostic accuracy was 86.1%.Conclusions: This study established a new method for the early detection of OLP by analyzing serum metabolomics using UHPLC-Q-Orbitrap HRMS, which will help in understanding the pathological processes of OLP and identifying precancerous lesions in oral cavity.

Trepibutone was widely used for cholelithiasis, cholecystitis, biliary tract dyskinesia, cholecystectomy syndrome, and chronic pancreatitis in clinic. However, few investigations on trepibutone have been conducted. In this study, an accurate, sensitive, and selective analytical method was developed and successfully applied to assess the pharmacokinetic behavior of trepibutone in rats. Trepibutone and carbamazepine (internal standard, IS) were quantified using multiple reaction monitoring (MRM) mode with the transitions of m/z 311.09→265.08 and m/z 237.06→194.08, respectively. The linearity, precision, accuracy, extraction recovery, matrix effect, and stability of the established method were all excellent within acceptable range. A total of 30 metabolites were identified in plasma and urine by Q-Exactive high resolution mass spectrometry, and several common metabolic pathways were observed such as dealkylation, oxidation, reduction, glucuronidation, and so on. This research provides more information on trepibutone in pharmacodynamics and toxicology and will assist the usage of trepibutone in clinical.

Safflower injection (SI) is a herbal injection made from the dried flower of Carthamus tinctorius L. SI is mainly used for the treatment of cerebral infarction, coronary heart disease, and other diseases. However, the active compounds responsible for the bioactivity of the SI remain unknown. In this study, a total of 46 compounds were identified in the SI by the ultra-high-performance liquid chromatography-Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry, and the molecular docking analysis showed that six compounds were probably served as the key components for the therapeutic effect of the SI against cerebral ischemia and coronary heart disease. Although the active compounds for the two diseases were same, the corresponding targets were different. The obtained results were useful for the quality control and the explanation of the pharmacological mechanism of the SI. The docking data were further subjected to the multivariate analysis, and the result showed that the active compounds were clustered together in the scatter plot, and the key targets could be determined by the VIP scores. Thus, the multivariate analysis could be used as a useful method to screen the active compounds and targets efficiently based on the docking data sets. [ABSTRACT FROM AUTHOR]

Trepibutone was widely used for cholelithiasis, cholecystitis, biliary tract dyskinesia, cholecystectomy syndrome, and chronic pancreatitis in clinic. However, few investigations on trepibutone have been conducted. In this study, an accurate, sensitive, and selective analytical method was developed and successfully applied to assess the pharmacokinetic behavior of trepibutone in rats. Trepibutone and carbamazepine (internal standard, IS) were quantified using multiple reaction monitoring (MRM) mode with the transitions of m/z 311.09→265.08 and m/z 237.06→194.08, respectively. The linearity, precision, accuracy, extraction recovery, matrix effect, and stability of the established method were all excellent within acceptable range. A total of 30 metabolites were identified in plasma and urine by Q-Exactive high resolution mass spectrometry, and several common metabolic pathways were observed such as dealkylation, oxidation, reduction, glucuronidation, and so on. This research provides more information on trepibutone in pharmacodynamics and toxicology and will assist the usage of trepibutone in clinical. [ABSTRACT FROM AUTHOR]

• Protein-peptide interaction, proteomics, computer molecular simulations were utilized. • mDIA acquisition mode demonstrated better performance than ddMS2 and vDIA modes. • Hydrogen bonds and hydrophobic interactions contributed to stability of HPFLEWAR. • HPFLEWAR mainly interacted with Lys69, Ile72, Asp53, and Glu74 of lactoglobulin. • Binding HPFLEWAR to β-lactoglobulin is effective way to mask bitterness perception. Suppressing the aversive bitterness of bioactive peptides is an arduous task as it hinders product acceptability. Three acquisition modes (ddMS2, vDIA, and mDIA) of high-resolution mass spectrometry (HRMS) were designed for structure confirmation and accurate quantification of HPFLEWAR, with the mDIA mode chosen as optimum. HRMS and isothermal titration calorimetry was used to elucidate the mechanism that β-lactoglobulin self-assemble to form association complex in 1:1 stoichiometric ratio (ΔG value − 29.36 kJ mol−1), which automatically attracted HPFLEWAR and reduces its distribution in free form, downgraded the level of bitter perception. Proteomics experiments and molecular dynamics simulations was built to discovered that HPFLEWAR bound and stabilized in the negatively charged region of β-lactoglobulin via four hydrogen bonds (Lys69, Ile72, Asp53, and Glu74) and hydrophobic interactions. These findings were considered to give theoretical foundation for strictly controlling the bitter perception of peptides and the possible application of HPFLEWAR as new functional components. [ABSTRACT FROM AUTHOR]

Flavor retention and consumer receptivity of goat milk are strongly influenced by the interaction between flavor constituents and proteins. Few previous works suggested the impact of interactions between β-lactoglobulin (β-LG) and flavor constituents in goat milk on the release of volatile compounds. In present research, binding of β-lactoglobulin with four model flavor constituents, octanoic acid (OA), 4-methyloctanoic acid (MCA), 2-tridecanone (TO) and tert-butanol (TB) was deciphered. UHPLC-Q-Orbitrap high-resolution mass spectrometry results identified that all flavor molecules bound to β-lactoglobulin in the 1:1 stoichiometric ratio. Docking analysis of the lowest binding energy indicated that the strongest binding relationships between β-LG and MCA/OA/TB/TO were expected to be β-LG/TO > β-LG/MCA > β-LG/OA > β-LG/TB, which were spontaneous. Via molecular dynamic simulations, TO was located at hydrophobic cavity of β-barrel causing β-LG structure to further curl. The spatial direction of MCA began to gradually approach outer surface near Trp19-Arg124. OA was located in entrance of the hydrophobic core of each β-barrel. TB remained relatively stable at surface hydrophobic sac in the groove between β-barrel and α-helix. The effect of β-LG conformation was also evaluated by fluorescence spectroscopy, which was same to the molecular dynamics simulation. Present study may help to better understand the interaction mechanism of β-LG and flavor constituents and evaluate the ability of flavor compounds having a broad array of different functional groups to react with β-LG, which provided theoretical basis for quality maintenance of goat milk. [Display omitted] • Noncovalent interaction of β-lactoglobulin/flavor constituents was investigated. • Hydrogen bond and hydrophobic contacts exist the β-lactoglobulin/flavor constituent. • 2-Tridecanone was located at hydrophobic cavity of β-barrel causing further curl. • Flavor constituents binding site remains rigid during molecular dynamics simulation. • β-Lactoglobulin was further compacted and had minor changes during docking process. [ABSTRACT FROM AUTHOR]

• LC–MS and NMR were integrated to identify the components of Shuxuening Injection. • Characteristic ions and MS network of flavonoids and TTLs were generated. • 133 compounds were identified, and the absence of toxic components was also confirmed. Shuxuening injection (SXNI) is made from the extract of Ginkgo biloba , and used for curing coronary heart disease, stenocardia and cerebral vasospasm. However, the flavonoids and terpene trilactones (TTLs) only account for about 30% of the dry weight of SXNI according to the current quality control standard. Thus, systematic investigation of the chemical composition of SXNI is needed to guarantee the drug safety. In this study, the integral approach of ultra high performance liquid chromatography-Q Exactive hybrid quadrupole-orbitrap high resolution accurate mass spectrometry (UHPLC-Q-Orbitrap HRMS) and nuclear magnetic resonance (NMR) techniques, combining with the solvent partition, were used for identifying the phytochemical compounds in SXNI. The NMR analysis revealed the possible structure types based on the characteristic chemical shifts, coupling constants, and peak shape, which gave the clues for the further UHPLC-Q-Orbitrap HRMS analysis of previously unknown or uncatalogued compounds. The characteristic ions and structural network of flavonoids and TTLs were generated and used in the UHPLC-Q-Orbitrap HRMS based structure identification. Only 72 compounds could be identified by the direct UHPLC-Q-Orbitrap HRMS analysis of SXNI, while this systematic strategy led to the identification of 133 compounds, which included 73 flavonoids, 5 biflavone, 6 proanthocyanidin, 14 TTLs, 6 fatty acids, 5 fatty amides, 12 organic acids, 1 amino acid, 3 excipients, 2 saccharides and 6 other components. In addition, the presence of steroids and triglycerides were also suggested by NMR. The absence of toxic components in SXNI was also confirmed. In addition, the NMR spectrum also revealed some unknown signals, which need to be further identified. This approach was proved to be useful for the rapid and effective identification of various compounds in the TCM preparations. [ABSTRACT FROM AUTHOR]

Curculigo orchioides and C. latifolia have been used as traditional medicines such as antidiabetic and anticancer. This study measured the total phenolics and flavonoid contents as well as analyzed the functional groups and chemical compounds using Fourier-transform infrared (FTIR) spectra and UHPLC-Q-Orbitrap-HRMS profiling for the discrimination of plant parts, geographical origin, and compounds that presumably have a significant contribution as antioxidant and α-glucosidase inhibitors on both plants. The total phenolics and flavonoids contents in Curculigo species varied from 142.09 to 452.47 mg gallic acid equivalent (GAE/g) and from 0.82 to 5.44 mg quercetin equivalent (QE/g), respectively. The lowest IC50 for antioxidant and α-glucosidase inhibitory activities is presented by C. latifolia from a higher altitude region. Principal component analysis (PCA) from FTIR and UHPLC-Q-Orbitrap-HRMS data could discriminate the plant parts and geographical origin. Partial least squares (PLS) analysis has identified several functional groups, such as O–H, C–H, C=O, C–C, C–O, and chemical compounds, unknown-185 and unknown-85, that are most likely to contribute to the antioxidant and α-glucosidase inhibitory activities.