Ultra-performance Liquid Chromatography-mass Spectrometer Research Articles

Comprehensive insights into the mechanism of flavor formation driven via inoculation with mixed starter cultures in dry-fermented tilapia sausages: Integration of macrogenomics, volatilomics, and lipidomics

To investigate the mechanisms of flavor formation in dry-fermented tilapia sausages, the volatiles, bacterial community, and lipid composition during fermentation were analyzed using gas chromatography-ion mobility spectrometry, 16S high throughput sequencing, and ultra-performance liquid chromatography-mass spectrometer. Pediococcus pentosaceus, Staphylococcus xylosus, and Staphylococcus carnosus became dominant bacteria during the fermentation. A total of 66 volatiles and 293 lipids (48 differential lipids) were identified. PC and PE content decreased. Aldehyde and 1-octen-3-ol content decreased. Most esters and ketones content increased during fermentation. Six metabolic pathways associated with differential lipids were identified by enrichment analysis. Glycerophospholipid metabolism was the main metabolic pathway. Correlation analysis revealed that PC and PE were precursors for volatiles, including PC 16:0/18:2 and PE 18:0/22:6. The dominant bacteria facilitate the hydrolysis of PC and PE, leading to the formation of esters and ketones. This study provides a theoretical basis for the targeted regulation of fermented sausage flavors.

Reaction mechanism, degradation pathway and toxicity assessment of NH4+ enhanced potassium ferrate removal of levofloxacin

Considering that ammonium ion (NH4+) was widely distributed in natural water and many sewage, it was of great practical significance to study NH4+ enhanced potassium ferrate (Fe(VI)) to rapid removal of levofloxacin (LEV) from water. In this experiment, firstly, the reaction kinetic of NH4+ enhanced Fe(VI) removal of LEV was studied, and the results showed that the enhanced removal reaction conformed to the second-order kinetic equation. When pH ≤ 9, the second-order rate constant kapp was linearly related to NH4+ dosage, but when pH= 10, the above linear relationship was broken, and the slope of the fitting line would change toward a smaller trend. Subsequently, tert-butanol (TBA) and Methyl phenyl sulfoxide (PMSO) were used as probes for·OH and high-valent iron-based intermediate (Fe(V)/Fe(IV)) respectively, which further revealed the reaction mechanism that NH4+ enhanced Fe(VI) to remove LEV as follows: NH4+ could promote Fe(VI) to produce a large amount of Fe(V)/Fe(IV), thus accelerating the degradation of LEV. In addition, 10 degradation intermediates of LEV were identified by quadrupole time-of-flight tandem ultra-performance liquid chromatography mass spectrometer (Q-TOF LC-MS), and then combined with density functional theory (DFT) calculation, two possible degradation pathways were inferred. Through the luminescence inhibition experiment of Vibrio fischeri and the toxicity evaluation of ECOSAR, it could be concluded that in the process of NH4+ enhancing Fe(VI) to remove LEV, the toxicity of the solution showed a trend of increasing first and then decreasing, and its toxicity was effectively improved and reduced after full oxidation. Finally, the experiments in actual water showed that it was feasible to use NH4+ in actual water to enhance Fe(VI) for rapid removal of LEV.

Peroxydisulphate activated FTO-WO3 nanorods based photoelectrocatalytic degradation of tetracycline: Intermediate products, degradation pathway and ecotoxicity studies

This work reports sulphate radical assisted photoelectrocatalytic (SR-PEC) degradation of tetracycline using a visible light active fluorine-doped tin oxide – tungsten trioxide nanorods (FTO-WO3 NRs) photoanode. The WO3 NRs were synthesised via the hydrothermal method and then conducted on the FTO glass to form a photoanode. When the photoanode was applied without sulphate radicals for PEC degradation, 10 % of the tetracycline was degraded. Conversely, when 3 mM persulphate was added, the extent of tetracycline degraded was 88 % using the UV–vis spectrophotometer and 99 % using the ultra-performance liquid chromatography mass spectrometer (UPLC-MS) within 90 min at 1.5 V. The mechanism of tetracycline degradation was proposed based on the intermediate products identified using UPLC-MS and the extent of toxicity was evaluated using quantitative structure activity relationship (QSAR) analysis. Trapping experiment revealed that the photogenerated holes, sulphate radicals, and hydroxyl radicals were the oxidants that significantly took part in the degradation of tetracycline. Overall, the electrode was stable and reusable, therefore suggesting the suitability of FTO-WO3 NRs photoanode in the presence of sulphate radicals towards the decontamination of water laden with pharmaceutical pollutants.

Saikosaponins regulate bile acid excretion in mice liver and ileum by activating farnesoid X receptor and bile acid transporter.

Radix Bupleuri exerts effective hepatoprotective and cholagogic effects through its Saikosaponins (SSs) component. Therefore, we attempted to determine the mechanism of saikosaponins used to promote bile excretion by studying their effects on intrahepatic bile flow, focusing on the synthesis, transport, excretion, and metabolism of bile acids. C57BL/6N mice were continuously gavaged with saikosaponin a (SSa), saikosaponin b2 (SSb2 ), or saikosaponin D (SSd) (200 mg/kg) for 14 days. Liver and serum biochemical indices were determined using Enzyme-linked immunosorbent assay (ELISA) kits. In addition, an ultra-performance liquid chromatography-mass spectrometer (UPLC-MS) was used to measure the levels of the 16 bile acids in the liver, gallbladder, and cecal contents. Furthermore, SSs pharmacokinetics and docking between SSs and farnesoid X receptor (FXR)-related proteins were analyzed to investigate the underlying molecular mechanisms. Administration of SSs and Radix Bupleuri alcohol extract (ESS) did not cause significant changes in alanine aminotransferase (ALT), aspartate aminotransferase (AST), or alkaline phosphatase (ALP) levels. Saikosaponin-regulated changes in bile acid (BA) levels in the liver, gallbladder, and cecum were closely related to genes involved in BA synthesis, transport, and excretion in the liver. Pharmacokinetic studies indicated that SSs were characterized by rapid elimination (t1/2 as 0.68-2.47 h), absorption (Tmax as 0.47-0.78 h), and double peaks in the drug-time curves of SSa and SSb2 . A molecular docking study revealed that SSa, SSb2 , and SSd docked well with the 16 protein FXR molecules and target genes (<-5.2 kcal/mol). Collectively, saikosaponins may maintain BA homeostasis in mice by regulating FXR-related genes and transporters in the liver and intestine.

Efficient removal of tetracycline in water by a Fe3O4-mediated persulfate activation and biodegradation coupled system: Performance, validation, and mechanism

The cost-effective removal of antibiotic contaminants from the aqueous environment has become a hot research topic now. In this study, we proposed a novel coupled system, i.e., nano-Fe3O4 (nFe3O4) mediated persulfate (PDS) activation and biodegradation process, for efficient removal of tetracycline (TCH). The feasibility of the coupled system (nFe3O4 + PDS + microbe) was proved by comparing it with a single AOP (nFe3O4 + PDS) in batch and column experiments. The highest removal efficiencies of TCH (20 mg/L) and TOC reached 99.5 % and 43.2 % by the coupled system within 60 days. SEM spectra exhibited that nFe3O4 and microorganisms were well loaded on the non-woven fabric. Meanwhile, XPS results proved that more Fe(II) was on the surface of nFe3O4 in the coupled system than that in the single AOP. The presence of abundant Fe(II) in the coupled system suggested the potential electron transfer from Fe(III) with the assistance of microbes. High-throughput sequencing results indicated Proteobacteria played an important role in the coupled system. The degradation intermediates of TCH were analyzed by ultraperformance liquid chromatography-mass spectrometer (UPLC-MS), and reasonable degradation pathways were proposed. The coupling of nFe3O4 + PDS and biological treatment improved the TCH removal efficiency and reduced the burden of subsequent water treatment, which provided a feasible idea for the treatment of antibiotic-containing wastewater.

UPLC-MS based integrated plasma proteomic and metabolomic profiling of TSC-RAML and its relationship with everolimus treatment.

Aim: To profile the plasma proteomics and metabolomics of patients with renal cysts, sporadic angiomyolipoma (S-AML) and tuberous sclerosis complex related angiomyolipoma (TSC-RAML) before and after everolimus treatment, and to find potential diagnostic and prognostic biomarkers as well as reveal the underlying mechanism of TSC tumorigenesis. Materials and Methods: We retrospectively measured the plasma proteins and metabolites from November 2016 to November 2017 in a cohort of pre-treatment and post-treatment TSC-RAML patients and compared them with renal cyst and S-AML patients by ultra-performance liquid chromatography-mass spectrometer (UPLC-MS). The tumor reduction rates of TSC-RAML were assessed and correlated with the plasma protein and metabolite levels. In addition, functional analysis based on differentially expressed molecules was performed to reveal the underlying mechanisms. Results: Eighty-five patients with one hundred and ten plasma samples were enrolled in our study. Multiple proteins and metabolites, such as pre-melanosome protein (PMEL) and S-adenosylmethionine (SAM), demonstrated both diagnostic and prognostic effects. Functional analysis revealed many dysregulated pathways, including angiogenesis synthesis, smooth muscle proliferation and migration, amino acid metabolism and glycerophospholipid metabolism. Conclusion: The plasma proteomics and metabolomics pattern of TSC-RAML was clearly different from that of other renal tumors, and the differentially expressed plasma molecules could be used as prognostic and diagnostic biomarkers. The dysregulated pathways, such as angiogenesis and amino acid metabolism, may shed new light on the treatment of TSC-RAML.

An optimal medicinal and edible Chinese herbal formula attenuates particulate matter-induced lung injury through its anti-oxidative, anti-inflammatory and anti-apoptosis activities

ObjectiveIdentifying novel strategies to prevent particulate matter (PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases. The combined intervention represented by herbal formulae for simultaneously targeting multiple pathological processes can provide a more beneficial effect than the single intervention. The aim of this paper is therefore to design a safe and effective medicinal and edible Chinese herbs (MECHs) formula against PM-induced lung injury. MethodsPM-induced oxidative stress, inflammatory response and apoptosis A549 cell model were used to screen anti-oxidant, anti-inflammatory and anti-apoptotic MECHs, respectively. A network pharmacology method was utilized to rationally design a novel herbal formula. Ultra performance liquid chromatography-mass spectrometer was utilized to assess the quality control of MECHs formula. The excretion of magnetic iron oxide nanospheres of the MECHs formula was estimated in zebrafish. The MECH formula against PM-induced lung injury was investigated with mice experiments. ResultsFive selected herbs were rationally designed to form a new MECH formula, including Citri Exocarpium Rubrum (Juhong), Lablab Semen Album (Baibiandou), Atractylodis Macrocephalae Rhizoma (Baizhu), Mori Folium (Sangye) and Polygonati Odorati Rhizoma (Yuzhu). The formula effectively promoted the magnetic iron oxide nanospheres excretion in zebrafish. The mid/high dose formula significantly prevented PM-induced lung damage in mice by enhancing the activity of SOD and GSH-Px, reducing the MDA and ROS level and attenuating the upregulation of pro-inflammatory cytokine (IL-6, IL-8, IL-1β and TNF-α), down regulating the protein expression of NF-κB, STAT3 and Caspase-3. ConclusionOur findings suggest that the effective MECHs formula will become a novel strategy for preventing PM-induced lung injury and provide a paradigm for the development of functional foods using MECHs.

A newly-integrated FeCo-layered double hydroxides photocatalytic system for UV-induced degradation of various heterocyclic amines against complex sample matrix

For several decades, toxic thermally processed hazards, represented by heterocyclic amines (HCAs), have gradually become a challenge in the field of purification owing to their increasing exposure, wide variety, and the complexity of the matrix. In this study, the first example of degradation-removal of HCAs was illustrated by using a newly-integrated Fe/Co co-doped layered double hydroxide (FeCo-LDH) as photocatalysts based on sulfate radical advanced oxidation processes (SR-AOPs). Under the excitation of UV light, a FeCo-LDH/sodium persulfate (PS) photocatalytic degradation system combined with ultra-performance liquid chromatography mass spectrometer (UPLC-MS) was established. The synthesized photocatalysts were optimized by varying the Fe/Co ratio, and a series of photochemical activities and reaction conditions were investigated for a better performance. As a result, the FeCo-LDH/PS system was able to degrade 12 kinds of common HCAs efficiently and simultaneously, and was also well applicable in real edible oil samples with completely degraded. Moreover, it exhibited well reusability, preserving 97.75% of initial activity after being used for 5 cycles. It is the first application of photocatalytic combined with SR-AOPs technology for the degradation of HCAs and provides a promising example for the regulation of thermally processed hazards.

Astragalus membranaceus ultrafine powder alleviates hyperuricemia by regulating the gut microbiome and reversing bile acid and adrenal hormone biosynthesis dysregulation

Metabolic syndrome is closely related to the intestinal microbiota and disturbances in the host metabolome. Hyperuricemia (HUA), a manifestation of metabolic syndrome, can induce various cardiovascular diseases and gout, seriously affecting a patient’s quality of life. Astragalus membranaceus has a long history as a commonly used traditional Chinese medicine to treat kidney disease in China and East Asia. We compared the therapeutic effect of benzbromarone and two different doses Astragalus membranaceus ultrafine powder (AMUP) in rats with HUA. Ultra-performance liquid chromatography-mass spectrometer was used to analyze the AMUP metabolism in the plasma, urine, and feces. Further, 16S ribosome RNA sequencing and feces metabolomic were performed to capture the variation of the gut microbiota and metabolites changes before and after drug administration. AMUP had a notable impact on reducing blood uric acid levels while protecting the liver and kidney. Drug metabolism analysis demonstrated that effective constituent flavonoids are distributed in the blood, whereas saponins remain in the intestine. Gut microbiota analysis showed that low-dose AMUP ameliorated HUA-induced gut dysbiosis by reducing the abundance of harmful bacteria and increasing that of some beneficial bacteria with anti-inflammatory properties, such as Clostridia, Lachnospiraceae, and Muribaculaceae. In addition, HUA-induced changes in metabolite contents in bile acid and adrenal hormone biosynthesis pathways were restored after treatment with AMUP. Low-dose AMUP exerts remarkable therapeutic effects on HUA by regulating the gut microbiome and mediating gut metabolism pathways associated with uric acid excretion.

Plasma metabolomic profiles reveal regulatory effect of chitosan oligosaccharides on loperamide-induced constipation in mice

Chitosan oligosaccharides (COS) can improve the symptoms of constipation. In this study, we further explored the regulator effect of COS on aberrant plasma metabolomics in constipated mice. Using untargeted metabolomic analysis by ultra-performance liquid chromatography-mass spectrometer (UPLC-MS), we identified several most significantly changed metabolic pathways in plasma of constipated mice induced by loperamide, including those correlated with the metabolisms of sphingolipid, glycerophospholipid, tryptophan, bile acids, unsaturated fatty acids, and amino acids. The changes in these metabolic pathways were reversed by COS treatment largely. Furthermore, the mRNA levels of some key target genes related to the above metabolic pathways in colon samples were detected by reverse transcription-polymerase chain reaction analysis. We showed that COS significantly suppressed the abnormal expression of these genes, including ceramide glucosyltransferase (CGT), sphingolipid 4-desaturase (DEGS2), alkaline ceramidase (ACER1), sphingosine kinase 2 (SPHK2), lysophosphatidylcholine acyltransferase (LPCAT1), and aromatic-L-amino-acid (DDC). These data provide insight into the mechanisms by which COS ameliorates loperamide-induced constipation in mice.

Dysregulated Metabolites Serve as Novel Biomarkers for Metabolic Diseases Caused by E-Cigarette Vaping and Cigarette Smoking.

Metabolites are essential intermediate products in metabolism, and metabolism dysregulation indicates different types of diseases. Previous studies have shown that cigarette smoke dysregulated metabolites; however, limited information is available with electronic cigarette (e-cig) vaping. We hypothesized that e-cig vaping and cigarette smoking alters systemic metabolites, and we propose to understand the specific metabolic signature between e-cig users and cigarette smokers. Plasma from non-smoker controls, cigarette smokers, and e-cig users was collected, and metabolites were identified by UPLC-MS (ultra-performance liquid chromatography mass spectrometer). Nicotine degradation was activated by e-cig vaping and cigarette smoking with increased concentrations of cotinine, cotinine N-oxide, (S)-nicotine, and (R)-6-hydroxynicotine. Additionally, we found significantly decreased concentrations in metabolites associated with tricarboxylic acid (TCA) cycle pathways in e-cig users versus cigarette smokers, such as d-glucose, (2R,3S)-2,3-dimethylmalate, (R)-2-hydroxyglutarate, O-phosphoethanolamine, malathion, d-threo-isocitrate, malic acid, and 4-acetamidobutanoic acid. Cigarette smoking significant upregulated sphingolipid metabolites, such as d-sphingosine, ceramide, N-(octadecanoyl)-sphing-4-enine, N-(9Z-octadecenoyl)-sphing-4-enine, and N-[(13Z)-docosenoyl]-sphingosine, versus e-cig vaping. Overall, e-cig vaping dysregulated TCA cycle-related metabolites while cigarette smoking altered sphingolipid metabolites. Both e-cig and cigarette smoke increased nicotinic metabolites. Therefore, specific metabolic signatures altered by e-cig vaping and cigarette smoking could serve as potential systemic biomarkers for early pathogenesis of cardiopulmonary diseases.

Efficient removal of tetracycline in water by a novel chemical and biological coupled system with non-woven cotton fabric as carrier

As a novel wastewater treatment strategy, the intimate coupling of photocatalysis and biodegradation (ICPB) has been attracted attention, which is ascribed to its combination of the advantages of photocatalytic reactions and biological treatment. The selection of carriers is important since it affects the stability of the system and the removal efficiency of pollutants. In this study, a novel ICPB system was successfully constructed by loading photocatalytic materials (i.e., TiO2, N-TiO2, and Ag-TiO2) and microbes onto non-woven cotton fabric. The photocatalysts were characterized by scanning electron microscope-energy dispersive spectrometer (SEM-EDS), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). This system exhibited good performance in degrading tetracycline (TC) in water. The results showed that Ag-TiO2-ICPB had the maximum removal efficiency of tetracycline (94.7%) in 5 h, which was 16.5% higher than the photocatalysis alone. After five cycles, 82.9% of tetracycline could be still degraded through Ag-TiO2-ICPB. SEM spectrum showed microbes on the material changed little before and after the reactions. This result implied the materials were stable, and then beneficial for degrading of pollutants continuously. The intermediates were detected through ultraperformance liquid chromatography-mass spectrometer (UPLC-MS) and the plausible degradation pathways were proposed. Electron paramagnetic resonance (EPR) analysis showed OH and O2− were the main reactive oxygen species for TC degradation. In conclusion, the ICPB system with non-woven cotton fabric as a carrier has certain application prospects for antibiotic-containing wastewater.

The protective effect of Veronica ciliata Fisch. Extracts on relieving oxidative stress-induced liver injury via activating AMPK/p62/Nrf2 pathway

Ethnopharmacological relevanceVeronica ciliata Fisch. existed in various Tibetan medicine prescriptions, which was recorded to treat liver diseases in the Tibetan medicine roll of Chinese materia medica. Hypothesis/purposeThe current study aimed to examine the effect of active constituents from V.ciliata relieving oxidative stress-mediated liver injury and clarify the underlying mechanism. Materials and methodstert-Butyl hydroperoxide (BHP) induced liver injury in mice model was established to evaluate the hepatoprotective effect of ethyl acetate extract of V. ciliata (EAFVC). Serum and liver indicators, as well as the histopathological change of liver were examined. Next, the constituents of EAFVC were separated and characterized by high-speed countercurrent chromatography (HSCCC) and Ultra performance liquid chromatography-mass spectrometer (UPLC-MS), respectively. Based on the above, the antioxidant activity of EAFVC and two fractions was evaluated using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2, 2'-azino-bis (3-ethylbenzothiazoli- ne-6-sulfonic acid) (ABTS) free radical scavenging assays. The hepatoprotective activity of EAFVC and its fractions/compounds attenuating ethanol-induced hepatocyte damage in BRL-3A cells was evaluated using the MTT method. The effect of the fraction and compounds with the strongest protective activity on ethanol-induced cytotoxicity, reactive oxygen species (ROS) accumulation, and glutathione (GSH) depletion was investigated. mRNA expression of nuclear factor-E2-related factor 2 (Nrf2) and nuclear factor of κB (NF-κB), as well as their downstream target genes, was determined by RT-qPCR. Finally, the potential mechanism of fraction 1 and luteolin on the AMPK/p62/Nrf2 signal pathway was studied using western blotting. ResultsFirstly, EAFVC could relieve liver impairment induced by t-BHP in mice. Next, fraction 1 enriched with polyphenolic compounds and luteolin derived from EAFVC were screened to yield the highest hepatoprotective activity against ethanol-induced hepatocyte damage. Further study demonstrated that fraction 1 and luteolin relieved BRL-3A cells damage by decreasing the aspartate aminotransferase (AST), alanine transaminase (ALT) and lactate dehydrogenase (LDH) activities, ROS accumulation, as well as the depletion of GSH. Also, we determined that fraction 1 and luteolin suppressed inflammation and apoptosis of BRL-3A cells. The mechanistic studies indicated that fraction 1 could attenuate oxidative stress, inflammation, and apoptosis by activating AMPK phosphorylation, which promotes autophagy associated protein expression (LC3-B, Beclin1 and p62) as well as promote phosphorylation of p62 -dependent autophagic degradation of Keap1, to induce Nrf2 dissociation from Keap1 and translocate to nuclear. Nrf2 in the nuclear activate cytoprotective related genes to exert hepatoprotective function. Finally, we found that luteolin activated the protein expression of p-AMPK, p-p62, p62, Nrf2, and its downstream target genes. ConclusionsThis study clarified that fraction 1 enriched phenolic compounds could attenuate ethanol-induced liver injury in BRL-3A cells via activating AMPK/p62/Nrf2 pathway. Luteolin could serve as the major bioactive component in the therapeutic effect of fraction 1. These active constituents in V. ciliata could be used as the potential drugs targeted activation of AMPK or p62 for relieving oxidative stress-mediated liver disorders.

Quality Parameters, Caffeine and Theobromine Contents and Antioxidant Activity of Artisan and Commercial Chocolate from Brazil

Several types of chocolate were analysed: bitter, half-bitter, milk chocolate, 70% cocoa, white chocolate and cocoa seeds. Analysis of fat, insoluble solids, total phenols, theobromine and caffeine contents and antioxidant capacity were carried out to evaluate the quality of artisan and commercial chocolate products in Brazil. Fat contents and insoluble residues were similar to those reported in the literature. We develop a simple method by HPLC-DAD (High-Performance Liquid Chromatography with Diode-Array Detection) to quantify methylxanthines whose identification has been confirmed by UPLC- MS (Ultra performance liquid chromatography—mass spectrometer). The levels of phenols, caffeine and theobromine varied between types. White chocolate had no phenolics nor caffeine or theobromine. The ones with the highest phenolic contents were those with the highest cocoa mass content. The same happened with the antioxidant activity, which is correlated to the amount of phenolics, caffeine and theobromine, shown by the statistical analysis. It is the presence of cocoa that determines the antioxidant activity of chocolates. This work contributed to the confirmation of the properties of the chocolates, highlighting the importance of the artisan product. Chocolates are important for human health and are considered functional foods.

Chemical study, antioxidant activity, and genotoxicity and cytotoxicity evaluation of Ruellia angustiflora

Ruellia angustiflora (Acanthaceae) is known as flower-of-fire, and its leaves are traditionally employed to promote wound healing. This study was aimed at extracting and characterizing the chemical constituents of the extracts of R. angustiflora obtained by ultrasound-assisted and supercritical fluid extractions, and subsequently investigate their antioxidant potential and cyto-genotoxicity. The extract obtained by ultrasound (UAE-EtOH) was characterized by ultraperformance liquid chromatography-mass spectrometer (UPLC-MS), and the extract obtained via supercritical fluid (SFE-CO2) by gas chromatograph-mass spectrometer (GC-MS). The antioxidant potential was verified by the antiradical activity against the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), and the cyto-genotoxicity evaluation to test cell viability and DNA damage was performed in peripheral blood mononuclear cells (PBMC) cultures. The SFE-CO2 extract presented some fatty acids, triterpenes, tetraterpenes, tocopherols and phytosterols. The UAE-EtOH extract contained phenolic acids and flavonoids, and showed the highest antioxidant capacity. Neither extract was genotoxic or cytotoxic at the tested concentrations.

Serum metabolomic profiles reveal the impact of BuZangTongLuo formula on metabolic pathways in diabetic mice with hindlimb ischemia

Ethnopharmacological relevanceBuZangTongLuo Formula (BZTLF) was the decoction of eight traditional Chinese medicines including Astragalus membranaceus, Dioscorea opposita, Salvia miltiorrhiza, Scrophularia ningpoensis, Ophiopogon japonicus, Panax ginseng, Fritillariae cirrhosae and Whitmania pigra. This formula has been used as an effective remedy for treatment of diabetic ischemia clinically. Aim of the studyIn previous study, we have reported the therapeutic effect of BZTLF on diabetic vascular dysfunction. However, it remains obscure about the role of metabolic pathways in BZTLF-initiated improvement on hindlimb ischemia. Materials and methodsDiabetic mice with hindlimb ischemia were orally administrated with BZTLF by gavage. The serum samples were prepared for untargeted metabolomic analysis by ultra-performance liquid chromatography-mass spectrometer. The metabolic network was built by integrating metabolite data with the Gene Expression Omnibus (GEO) dataset (GSE3313). Further, quantitative PCR was used to confirm the key target genes. ResultsBZTLF treatment remarkably led to the reversal of changed metabolite levels in serum of diabetic mice with hindlimb ischemia, which mainly derived from bacteria, plant and signaling molecules. Also, BZTLF reshaped the metabolic pathways, especially those responsible for metabolism of lipid, gluthanine and tryptophan. In addition, BZTLF led to the reduction of lysophosphatidic acids (LPAs) and increment of triglycerides (TGs) conjugation with non-saturated fatty acids in serum. BZTLF significantly restored the down-regulation of vascular endothelial growth factor receptor 2 (VEGFR2) and endothelial nitric oxide synthase (eNOS) or the up-regulation of interleukin 4-induced 1 (IL4I1) and cytochrome P450 family 1 subfamily B member 1 (CYP1B1) at mRNA level, which were key regulatory genes located in metabolic pathways of glutamate and tryptophan. ConclusionsBZTLF improved hindlimb ischemia in diabetic mice by the positive regulation of metabolome changes in serum.

Metabolic profiling of coumarins by the combination of UPLC-MS-based metabolomics and multiple mass defect filter

Coumarins have aroused high interests due to their diverse bioactivities. Understanding of its metabolism contributes to determine the druggability of coumarin in vivo.A sensitive and efficient strategy based on ultra-performance liquid chromatography-mass spectrometer (UPLC-MS) analysis combined with various data-processing techniques including metabolomics and multiple mass defect filter (MMDF) was established for the comprehensive screening and elucidation of potential coumarin metabolites.Total 20 metabolites of scoparone were identified in this study, including 14 undescribed metabolites. The metabolism of two other similar coumarins scopoletin and esculetin also could be determined using this strategy.By the established strategy, this study gives the insights about the major metabolic pathways of scoparone in vivo and in vitro metabolism, including demethylation, hydroxylation, hydration, cysteine conjugation, glucuronide conjugation and sulfate conjugation. Additionally, the metabolic pathways of scopoletin and esculetin were determined as hydroxylation, glucuronidation and sulfation. These results contribute to the understanding of metabolic characterization of coumarins, and demonstrate that the combination of UPLC-MS-based metabolomics and MMDF is a powerful approach to determine the metabolic pathways of coumarin compounds.

Changes of 5-hydroxymethyl-2-furfural in fresh and processed ginsengs.

The study estimated changes of 5‐hydroxymethyl‐2‐furfuraldehyde (5‐HMF) in different ginseng products with different temperatures and time pretreatment. Heat treatment was performed at various temperatures for 1.50, 2.00, 2.50, and 3.00 hr, respectively. Ultrasonic extraction and reflux extraction were used to evaluate the extraction rate and different solvents (such as 80% methanol, dichloromethane, ethyl acetate, and an extraction with both dichloromethane and ethyl acetate solvents) using two extraction methods (liquid–liquid extraction and solid‐phase extraction) to remove matrix interference. An ultraperformance liquid chromatography–mass spectrometer (UPLC‐MS) method was used for quantitative and changing analysis of 5‐HMF in different ginseng samples. The results indicated that the content of 5‐HMF increased dramatically with heating temperature and time, and the 5‐HMF in the ginseng samples ranged from 0.01 to 112.32 g/kg protein. The highest value was observed in the honey‐added ginseng samples with the highest amount of addition and highest temperature treatment, and the lowest value was found in the fresh ginseng samples. These results implied that 5‐HMF may be as an indicator to estimate the honey addition level and heat treatment degree during the processing of ginseng products, and the content of 5‐HMF is a promising parameter to evaluate the quality of products (ginseng). The production and regulation of potentially harmful Maillard reaction products (PHMRPs)‐5‐HMF in ginseng manufacture will provide an important reference for safe ginseng processing.

Metabolomics and 16S rRNA Gene Sequencing Analyses of Changes in the Intestinal Flora and Biomarkers Induced by Gastrodia-Uncaria Treatment in a Rat Model of Chronic Migraine.

Accumulating evidence suggests that natural medicines have notable curative effects on neurological conditions, such as migraine, that are mediated by regulating the gut microbial flora. A natural medicine pair used in traditional Chinese medicine, Gastrodia elata Blume and Uncaria rhynchophylla (Miq.) Miq. ex Havil. (GU), have shown excellent effect in treating migraine, yet the role of gut microbes in the therapeutic effect of GU in chronic migraine (CMG) is unknown. Here, we performed a 16S rRNA gene sequencing and metabolomics study of the effects of GU in a nitroglycerin (NTG)-induced rat model of CMG. Our results showed that the gut microbial community structure changed significantly and was similar to that of control rats after GU administration in CMG rats. Specifically, GU increased the relative abundance of Bacteroides and Coprococcus and reduced the abundance of Prevotella_1 and Escherichia-Shigella in CMG rats. The metabolomics profiles of the plasma and ileum contents of CMG rats obtained with an ultra-performance liquid chromatography-mass spectrometer (UPLC-MS) revealed similar biomarkers in both samples, and GU treatment reduced 3-indoxyl sulfate, glutamic acid, L-tyrosine, and L-arginine levels, and increased 5-HIAA, L-tryptophan, and linoleic acid levels in plasma. Correlation analysis showed that the affected bacteria were closely related to amino acid metabolism. Most importantly, GU treatment hardly affected biomarkers in feces samples after inhibiting the activity of gut microbes. Collectively, these findings indicate that structural changes in gut flora are closely related to host metabolism and that regulating the gut microbial community structure and function may be one of the important mechanisms underlying the therapeutic effects of GU in migraine.

New insights into Paeoniaceae used as medicinal plants in China

Paeoniaceae is an abundant germplasm resource with significant medicinal values in China, the principal medicinal components of which include paeoniflorin and paeonol. These compounds are typically obtained from air-dried root samples, which the use of freeze-drying as an alternative method has not been tested. Additionally, the presence of these two compounds in various wild Paeoniaceae species has not been previously explored, nor have the differences between various plant organs been fully evaluated. Here, freeze-drying and air-drying methods were compared to assess the changes in paeoniflorin and paeonol in root samples using ultra-performance liquid chromatography-mass spectrometer. The contents of these compounds in the roots, leaves, stems, and petals were then tested in freeze-dried materials. We also quantitatively detected the paeoniflorin and paeonol contents in the roots of 14 species collected from 20 natural habitats. Results indicated that the paeoniflorin content decreased under air-drying in comparison to freeze-drying, while the opposite trend was observed for paeonol. Our findings also demonstrated that the root xylem of species in Section Moutan, particularly Paeonia ostii, contains considerable paeonol and paeoniflorin and should thus be fully utilized as a medicinal resource. Furthermore, paeonol was mainly detected in the roots, while paeoniflorin was widely distributed in different organs; the highest content was in the leaf at the budding stage, suggesting that the leaves should be developed as a new paeoniflorin resource. Paeoniflorin contents were also found to be higher at earlier development stages. Based on the standards of the Chinese Pharmacopoeia, five species of Section Moutan and six species of Section Paeonia could be used as potential traditional Chinese medicinal resources. These findings of this study enhance our understanding of these two medicinal compounds and provide a foundation for the further development and utilization of Paeoniaceae as medicinal plant resources.