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Objective:To investigate the anti-inflammatory effects of water extract of the<italic> Iris halophila</italic> root on lipopolysaccharide（LPS） stimulated RAW264.7 cells and analyze its chemical constituents. Method:The supernatant of YWG prepared by water extraction and alcohol precipitation was separated by AB-8 macroporous adsorption resin column chromatography to obtain ethanol eluates with different concentrations （YWG，YWG-0%，YWG-20%，YWG-40%，and YWG-60%）. Cell counting kit-8（CCK-8） assay was used to determine the effects of YWG-0%，YWG-20%，YWG-40%，and YWG-60% on the viability of RAW264.7 cells. Griess assay was employed to detect the nitric oxide （NO） level in LPS-stimulated RAW264.7 cells. The release of tumor necrosis factor（TNF）-<italic>α</italic>，interleukin（IL）-6，IL-10，and IL-1<italic>β</italic> was detected by enzyme-linked immunosorbent assay（ELISA）. YWG and the elution site with the most robust anti-inflammatory activity were identified and compared by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-Q-TOF-MS/MS）. Result:Ethanol eluates with different concentrations inhibited the release of NO，TNF-α，IL-1<italic>β</italic>， and IL-6 in the supernatant of LPS induced RAW264.7 cells （<italic>P<</italic>0.05），and promoted the release of IL-10 （<italic>P<</italic>0.05）. YWG-60% displayed a highly significant effect （<italic>P</italic><0.01）. A total of 127 constituents were detected from the comparison of YWG and YWG-60% by UHPLC-Q-TOF-MS/MS in the positive and negative ion modes，including 61 flavonoids. YWG-60% contained 25 flavonoids with elevated content as compared with YWG. Conclusion:YWG-60% showed potent anti-inflammatory effect，and the effective anti-inflammatory constituents were presumedly flavonoids. The findings of this study are expected to provide a scientific theoretical basis for the basic research on the medicinal effect of the water extract of YWG.
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Objective:To identify the chemical constituents of Platycladi Cacumen<italic> </italic>before and after being carbonized. Method:Chemical constituents in 3 batches of Platycladi Cacumen and its carbonized products<italic> </italic>were identified and compared by ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-QTOF-MS/MS）. Chromatographic separation was performed on an ACQUITY UPLC BEH C<sub>18</sub> column （2.1 mm×100 mm， 1.7 μm） with 0.1% formic acid aqueous solution （A）- acetonitrile （B） as mobile phase for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B； 18-22 min， 100%B）. The flow rate was 0.4 mL·min<sup>-1</sup> and the injection volume was 5 μL. Mass spectrometry was performed by an electrospray ionization， and the primary and secondary mass spectrometry data were collected with the full scan mode of positive and negative ions， the peaks containing MS/MS data were identified by self-established secondary mass spectrometry database and corresponding fragmentation law matching method. Result:A total of 77 and 76 substances with the same change trend were identified under positive and negative ion modes. After being<italic> </italic>carbonized， the disappeared components of Platycladi Cacumen were mainly amino acids， ketone aldehydes and other volatile components. Among newly produced components， there were 6 kinds of flavonoid aglycones （rhamnetin， 6，7，3'-trihydroxyflavone， 3，6，3'-trihydroxyflavone， 4'-hydroxy-2'-methyl-3，4，5-trimethoxychalcone， herbacetin and 3'，5'-dimethoxy-3，5，7，4'-tetrahydroxyflavone）， 3 kinds of coumarins （7-hydroxycoumarin， 7，8-dihydroxycoumarin and 8-acetyl-7-hydroxycoum-arin） and 3 kinds of benzoic acids （3-methylcatechol， pyrocatechol and chromone-3-carboxylic acid）. There were a total of 40 flavonoids （quercitrin， quercetin， kaempferol， etc.） among these identified chemical constituents. Conclusion:There are significant quantitative and qualitative changes in the chemical compositions of Platycladi Cacumen after being carbonized. The flavonoids， the identified main active ingredients， can provide data reference for further study on the material basis of efficacy changes of Platycladi Cacumen<italic> </italic>before and after being carbonized.
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The paper aims to compare the protective effect of Salvia miltiorrhiza and Anemarrhena asphodeloides on AD cell model and investigate its protective mechanism by cell metabolomics platform. AD cell model was established by the abnormal phosphorylation of Tau protein in SH-SY5Y cells induced by okadaic acid. The protective effect of the extract of Salvia miltiorrhiza and Anemarrhena asphodeloides on the model was evaluated by cell proliferation-toxicity experiment. The metabolomics platform was used to study the efficacy of Salvia miltiorrhiza and Anemarrhena asphodeloides comprehensively, explore the potential biomarkers related to AD and the effect of drugs on the potential biomarkers. Salvia miltiorrhiza extract had a certain protective effect on the AD model (P < 0.05), while the Anemarrhena asphodeloides extract had no significant protective effect (P > 0.05). 45 significant differential metabolites and the related 12 metabolic pathways were identified using UHPLC-QTOF/MS platform, which were related to the AD cell model. After administration of Salvia miltiorrhiza extract, 30 different metabolites appeared callback, while after intervention of Anemarrhena asphodeloides extract, 7 metabolites appeared callback. The results showed that the extracts of Salvia miltiorrhiza and Anemarrhena asphodeloides had certain protective effects on the AD cell model with Tau protein abnormal phosphorylation, but Salvia miltiorrhiza had more extensive targets and could significantly improve the cell viability. The mechanism may be related to the regulation of the metabolic pathways of AD cell model induced by okadaic acid.
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An ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) method was established to identify the metabolites in rat plasma after oral administration of Zhikebao tablets. The high-resolution mass spectrometer was operated in positive and negative ion mode, respectively. First, full-scan was applied, which was dependent on a multiple mass defect filter (MMDF) combined with dynamic background subtraction (DBS). These were utilized to trigger the information dependent acquisition (IDA) function in the experiment. For the IDA criteria, the eight most intense candidate ions of per cycle were selected to do a product ion scan. Then Metabolite Pilot 2.0 software was utilized to load data to seek possible metabolites. The analytical models employed by Metabolite pilot 2.0 were established for representative compounds of the Papaveris Pericarpium and licorice in Zhikebao tablet. Finally, metabolites were identified according to accurate mass measurement and retention time. 38 components from the rat plasma after oral administration of the drug have been found, including 5 prototype opium alkaloids, liquiritin, glycyrrhizic acid and 31 relative metabolites. The metabolic transformation of Zhikebao tablet in rats was mainly induced by glucuronidation, sulfation, methylation, amine to carboxylic acid, hydrolysis and so on. In this paper, the metabolites of the main active components of Zhikebao tablet were tentatively identified, and the metabolic pathway was compared with that of single chemical drugs. Moreover, it laid the fundamental elucidation of further metabolism study of Zhikebao tablet or other compound traditional Chinese medicine preparations which containing Papaveris Pericarpium or licorice.
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UHPLC-QTOF-MS was applied to non-targeted metabolomics study of mice infected with K. pneumoniae ATCC® BAA 2146 to discover potential biomarkers and metabolic pathways that are associated with sepsis. Fifty-eight metabolites were identified by principal components analysis (PCA) and partial least-squares discriminant analysis (OPLS-DA), which was combined with variable projection importance (VIP) and nonparametric test. Eighteen of the 58 metabolites were further found to be involved in 8 metabolic pathways, including nicotinate and nicotinamide metabolism, pyrimidine metabolism, vitamin B6 metabolism, taurine and hypotaurine metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, D-glutamine and D-glutamate metabolism and glycerophospholipid metabolism.
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Scopolin (SC-1), scopoletin (SC-2) and isofraxidin (IS-1) are the main active constituents in Chimonanthi Radix. However, the in vivo metabolism of SC-1, SC-2 and IS-1 have not been comprehensively clarified. In this study, the in vivo metabolic profiles of these three coumarins in the rat plasma, urine and feces were analyzed. Ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS/MS) method was applied to characterize the prototypes and metabolites of SC-1, SC-2 and IS-1 in rat feces, urine, and plasma after intravenous administration. A total of 11 metabolites of the three parent compounds were tentatively identified. The main metabolic pathways were analyzed by identification of metabolites, and it was found that these three coumarins underwent multiple in vivo metabolic reactions including glucuronidation, sulfonation, isomerism and reduction. In this study, the analysis of metabolites of three coumarins basically demonstrated their in vivo metabolic process, providing basis for the further pharmacokinetics and pharmacological evaluations of SC-1, SC-2 and IS-1.
Subject(s)
Animals , Rats , Calycanthaceae , Chemistry , Chromatography, High Pressure Liquid , Coumarins , Metabolism , Pharmacokinetics , Drugs, Chinese Herbal , Metabolism , Pharmacokinetics , Tandem Mass SpectrometryABSTRACT
Abstract Plants are a very rich source of pharmacologically relevant metabolites. However, the relative concentrations of these compounds are subject to the genetic make-up, the physiological state of the plant as well as environmental effects. Recently, metabolic perturbations through the use of abiotic stressors have proven to be a valuable strategy for increasing the levels of these compounds. Oxidative stress-associated stressors, including ionizing radiation, have also been reported to induce metabolites with various biological activities in plants. Hence, the aim of the current study was to investigate the effect of gamma radiation on the induction of purported anti-cancerous metabolites, glucomoringin and its derivatives, in Moringa oleifera Lam., Moringaceae. Here, an UHPLC-qTOF-MS-based targeted metabolic fingerprinting approach was used to evaluate the effect of gamma radiation treatment on the afore-mentioned health-beneficial secondary metabolites of M. oleifera. Following radiation, an increase in glucomoringin and three acylated derivatives was noted. As such, these molecules can be regarded as components of the inducible defense mechanism of M. oleifera as opposed to being constitutive components as it has previously been assumed. This might be an indication of a possible, yet unexplored role of moringin against the effects of oxidative stress in M. oleifera plants. The results also suggest that plants undergoing photo-oxidative stress could accumulate higher amounts of glucomoringin and related molecules.
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ABSTRACT Gentiana veitchiorum Hemsl., Gentianaceae, a traditional Tibetan medicine, was used for the treatment of liver jaundice with damp-heat pathogen, as well as for headache and chronic pharyngitis. A rapid ultra-performance liquid chromatography, photodiode array detector, quadrupole time-of-flight mass spectrometry method was developed for the fast and accurate identification and quantification of the chemical constituents of G. veitchiorum. In fact, eighteen compounds were detected and identified on the basis of their mass spectra, fragment characteristics and comparison with published data. Especially, the MS fragmentation pathways of iridoid glycosides and flavone C-glycosides were illustrated. Five compounds among them were quantified by UHPLC-PDA, including swertiamarin, gentiopicroside, sweroside, isoorientin, and isovitexin. The proposed method was then validated based on the analyses of linearity, accuracy, precision, and recovery. The overall recoveries for the five analytes ranged from 96.54% to 100.81%, with RSD from 1.05% to 1.82%. In addition, ten batches of G. veitchiorum from different areas were also analyzed. The developed method was rapid and reliable for both identification and quantification of the chemical constituents of G. veitchiorum, especially for simultaneous qualitative and quantitative analysis of iridoid glycosides and flavone C-glycosides.
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An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF/MS)method has been established to analyze the bioactive components of the aqueous extract of Euodiae Fructusand detect the metabolites in rats after oral administration of the aqueous extract. Compounds were detected by analyzing their retention times, high resolution mass data, mass spectra and comparing with the reference substances. As a result, 27 compounds were characterized from the aqueous extract of Euodiae Fructus, and 25 compounds were identified on the basis of the reference substances. 16 parent compounds and 35 metabolites were detected in rats' plasma, urine and feces samples, and all of the parent compounds were determined unambiguously through comparison with the reference substances. Four classes of compounds were identified from the aqueous extract of Euodiae Fructus, including phenolic acids, limonins, flavonoid glycosides and alkaloids. Phenolic acids, flavonoid glycosides and alkaloids were detected in rats' plasma, urine and feces, while limonins were detected in the urine and feces. The main metabolism pathways of these compounds in rats might include hydroxyl, hydrogenation, methylation, sulfating, glucuronidation, and so on. The results of this study provide references in the material basis of Euodiae Fructus in vivo.
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ABSTRACT Plants from the Momordica genus, Curcubitaceae, are used for several purposes, especially for their nutritional and medicinal properties. Commonly known as bitter gourds, melon and cucumber, these plants are characterized by a bitter taste owing to the large content of cucurbitacin compounds. However, several reports have shown an undisputed correlation between the therapeutic activities and polyphenolic flavonoid content. Using ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry in combination with multivariate data models such as principal component analysis and hierarchical cluster analysis, three Momordica species (M. foetida Schumach., M. charantia L. and M. balsamina L.) were chemo-taxonomically grouped based on their flavonoid content. Using a conventional mass spectrometric-based approach, thirteen flavonoids were tentatively identified and the three species were found to contain different isomers of the quercetin-, kaempferol- and isorhamnetin-O-glycosides. Our results indicate that Momordica species are overall very rich sources of flavonoids but do contain different forms thereof. Furthermore, to the best of our knowledge, this is a first report on the flavonoid content of M. balsamina L.
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A high throughput UHPLC-QTOF/MS method was established to analyze and identify the constituents in rat plasma after oral administration of honey-fried Stemona tuberosa Lour. extract. Automated data analysis software MetaboLynx was developed in analysis and identification of the bioactive components and their metabolites in rat plasma following oral administration of honey-fried S. tuberosa Lour. extract. As a result, 40 compounds including 12 prototype components and 28 metabolites were characterized. The constituents absorbed into blood and the possible metabolites which were demonstrated to originate from the active fraction of honey-fried S. tuberosa Lour. were responsible for the observed efficacy. Its serum pharmacochemistry should be subjected to complete investigation to illuminate the pharmacology and active mechanism of honey-fried S. tuberosa Lour.
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In present study, an ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS)-based plant metabolomics approach was established to investigate the metabolic profiles of the leaves, main root, branch root, and rhizome of Mountain Cultivated Ginseng (MCG). The UHPLC-QTOF/MS data were subjected to principal component analysis (PCA) and orthogonal partial least squared discrimination analysis (OPLS-DA) to find the potential characteristic components of the four parts of MCG in a quick way. The four different parts could be separated into four different groups of phytochemicals according to the PCA scores. The chemical constituents in four parts of MCG were obviously different. The identities of 81 major peaks that were detected in the four parts of MCG and the potential markers were identified by comparison with the reference compounds or were tentatively assigned by matching the retention time,empirical molecular formula and fragment ions with those of the published compounds of the Panax species. This proposed analytical method is fast, accurate, and reliable for differentiating the different parts of MCG. Moreover, this study supplied a new method for the quality evaluation of other Chinese medicinal materials.
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UHPLC-QTOF/MS technique was used to study the differences of lignans and their metabolites derived from Schisandra chinensis and vinegar Schisandra chinensis in rat plasma, bile, urine and faeces by the data processing techniques such as the dynamic background subtract (DBS), mass defect filtering (MDF) and enhance peak list (EPL) in analysis. In order to enhance accuracy for Schisandra chinensis hepatoprotective effect, we established rat acute alcoholic liver injury model in this experiment, and studied the prototype components and metabolisms of Schisandra lignans in vivo under pathological condition. The main ingredients of alcohol extract are lignans, including deoxyschizandrin, schisandrin B, schizandrin C, schizandrol, schizandrol B, schisantherin, schisantherin B, schisanhenol, gomisin G, gomisin J. The metabolic transformation of lignans in rats was mainly induced by methylation, hydroxyl, oxidation, and so on. Finally, we identified 6 kinds of prototype components and their 20 potential metabolites in Schisandra chinensis group and vinegar Schisandra chinensis group.