16 results on '"polar metabolites"'
Search Results
2. Optimization and Validation of a Method to Determine Enolones and Vanillin Derivatives in Wines—Occurrence in Spanish Red Wines and Mistelles
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Mónica Bueno, Julián Zapata, Laura Culleré, Ernesto Franco-Luesma, Arancha de-la-Fuente-Blanco, and Vicente Ferreira
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polar metabolites ,enolones ,vanillin derivatives ,sotolon ,furaneol ,vanillin ,wine ,mistelle ,odor activity value ,Chemistry (miscellaneous) ,Organic Chemistry ,Drug Discovery ,Molecular Medicine ,Pharmaceutical Science ,Physical and Theoretical Chemistry ,Analytical Chemistry - Abstract
Understanding the chemical nature of wine aroma demands accurate quantitative determinations of different odor-active compounds. Quantitative determinations of enolones (maltol, furaneol, homofuraneol, and sotolon) and vanillin derivatives (vanillin, methyl vanillate, ethyl vanillate, and acetovanillone) at low concentrations are complicated due to their high polarity. For this reason, this paper presents an improved and automated version for the accurate measure of these common trace wine polar compounds (enolones and vanillin derivatives). As a result, a faster and more user-friendly method with a reduction of organic solvents and resins was developed and validated. The optimization of some stages of the solid phase extraction (SPE) process, such as washing with an aqueous solution containing 1% NaHCO3 at pH 8, led to cleaner extracts and solved interference problems. Due to the polarity of these type of compounds, an optimization of the large volume injection was also carried out. Finally, a programmable temperature vaporization (PTV) quartz glass inlet liner without wool was used. The injector temperature was raised to 300 °C in addition to applying a pressure pulse of 180 kPa for 4 min. Matrix effects were solved by the use of adequate internal standards, such as ethyl maltol and 3′,4′-(methylenedioxy)acetophenone. Method figures of merit were highly satisfactory: good linearity (r2 > 0.98), precision (relative standard deviation, RSD < 10%), high recovery (RSD > 89%), and low detection limits ( 1) while homofuraneol and sotolon could also produce changes in their aroma perceptions (0.1 < OAV < 1).
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- 2023
3. Antibacterial activity and HPLC analysis of extracts from Mediterranean brown algae
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Rosa Alduina, Antonio Palumbo Piccionello, Elisabetta Oddo, Antonella Maggio, Anna Maria Mannino, Maggio A., Alduina R., Oddo E., Palumbo Piccionello A., and Mannino A.M.
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Mediterranean climate ,methanolic extract ,Plant Science ,Biology ,Cystoseira ,brown algae ,Settore BIO/19 - Microbiologia Generale ,01 natural sciences ,03 medical and health sciences ,Sensu ,Botany ,Mediterranean Sea ,Settore BIO/04 - Fisiologia Vegetale ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,0303 health sciences ,Hplc analysis ,010405 organic chemistry ,Settore BIO/02 - Botanica Sistematica ,Settore CHIM/06 - Chimica Organica ,biology.organism_classification ,phenol compound ,polar metabolites ,0104 chemical sciences ,Brown algae ,Antibacterial activity - Abstract
The antibacterial activity of methanolic extracts of eight Mediterranean brown algae belonging to the genera Cystoseira sensu lato, Dictyopteris and Padina was investigated. Extracts from Sargassaceae showed antibacterial activity. Specifically, Carpodesmia crinita and Carpodesmia brachycarpa extracts inhibited the growth of the Gram-positive strain Kocuria rhizophila whereas Cystoseira compressa and Carpodesmia amentacea extracts showed antibacterial activity against the Gram-positive strains, Kocuria rhizophila and Staphylococcus aureus. None of them inhibited the growth of the Gram-negative bacterium Escherichia coli. The extracts of Cystoseira sp. pl. and Treptacantha ballesterosii did not show any antibacterial activity. HPLC-MS analysis of the extracts allowed identification of polar compounds. Taxa of the genus Cystoseira sensu lato showed a higher number of compounds than Dictyopteris poly- podioides and Padina pavonica. Some differences found among and within taxa can be attributed to the different depths where these algae live, to thallus morphology and herbivore pressure. Particularly interesting were the phenol compounds, which showed differences among the genera but also within the genus Cystoseira sensu lato. Noteworthy, the almost exclusive presence of the fukiik acid isomer and of bromo-phloroglucinol in Carpodesmia amentacea, and the presence of exifone, able to bind iron metal ions, in Carpodesmia amentacea, Cystoseira compressa and Treptacantha ballesterosii.
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- 2020
4. Differential Cell Metabolic Pathways in Gills and Liver of Fish (White Seabream Diplodus sargus) Coping with Dietary Methylmercury Exposure
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Giuseppe De Marco, Barbara Billè, Fátima Brandão, Mariachiara Galati, Patrícia Pereira, Tiziana Cappello, and Mário Pacheco
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organic mercury ,dietary exposure ,gills ,Chemical Health and Safety ,white seabream ,total Hg accumulation ,Health, Toxicology and Mutagenesis ,NMR-based metabolomics ,fish metabolome ,liver ,Toxicology ,polar metabolites - Abstract
Mercury (Hg) is a dangerous and persistent trace element. Its organic and highly toxic form, methylmercury (MeHg), easily crosses biological membranes and accumulates in biota. Nevertheless, understanding the mechanisms of dietary MeHg toxicity in fish remains a challenge. A time-course experiment was conducted with juvenile white seabreams, Diplodus sargus (Linnaeus, 1758), exposed to realistic levels of MeHg in feed (8.7 μg g−1, dry weight), comprising exposure (E; 7 and 14 days) and post-exposure (PE; 28 days) periods. Total Hg levels increased with time in gills and liver during E and decreased significantly in PE (though levels of control fish were reached only for gills), with liver exhibiting higher levels (2.7 times) than gills. Nuclear magnetic resonance (NMR)-based metabolomics revealed multiple and often differential metabolic changes between fish organs. Gills exhibited protein catabolism, disturbances in cholinergic neurotransmission, and changes in osmoregulation and lipid and energy metabolism. However, dietary MeHg exposure provoked altered protein metabolism in the liver with decreased amino acids, likely for activation of defensive strategies. PE allowed for the partial recovery of both organs, even if with occurrence of oxidative stress and changes of energy metabolism. Overall, these findings support organ-specific responses according to their sensitivity to Hg exposure, pointing out that indications obtained in biomonitoring studies may depend also on the selected organ.
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- 2023
5. Intrapersonal Stability of Plasma Metabolomic Profiles over 10 Years among Women
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Oana A. Zeleznik, Clemens Wittenbecher, Amy Deik, Sarah Jeanfavre, Julian Avila-Pacheco, Bernard Rosner, Kathryn M. Rexrode, Clary B. Clish, Frank B. Hu, and A. Heather Eliassen
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Endocrinology, Diabetes and Metabolism ,lipids and lipid-related metabolites ,polar metabolites ,within-person stability ,unknown metabolite features ,Molecular Biology ,Biochemistry - Abstract
In epidemiological studies, samples are often collected long before disease onset or outcome assessment. Understanding the long-term stability of biomarkers measured in these samples is crucial. We estimated within-person stability over 10 years of metabolites and metabolite features (n = 5938) in the Nurses’ Health Study (NHS): the primary dataset included 1880 women with 1184 repeated samples donated 10 years apart while the secondary dataset included 1456 women with 488 repeated samples donated 10 years apart. We quantified plasma metabolomics using two liquid chromatography mass spectrometry platforms (lipids and polar metabolites) at the Broad Institute (Cambridge, MA, USA). Intra-class correlations (ICC) were used to estimate long-term (10 years) within-person stability of metabolites and were calculated as the proportion of the total variability (within-person + between-person) attributable to between-person variability. Within-person variability was estimated among participants who donated two blood samples approximately 10 years apart while between-person variability was estimated among all participants. In the primary dataset, the median ICC was 0.43 (1st quartile (Q1): 0.36; 3rd quartile (Q3): 0.50) among known metabolites and 0.41 (Q1: 0.34; Q3: 0.48) among unknown metabolite features. The three most stable metabolites were N6,N6-dimethyllysine (ICC = 0.82), dimethylguanidino valerate (ICC = 0.72), and N-acetylornithine (ICC = 0.72). The three least stable metabolites were palmitoylethanolamide (ICC = 0.05), ectoine (ICC = 0.09), and trimethylamine-N-oxide (ICC = 0.16). Results in the secondary dataset were similar (Spearman correlation = 0.87) to corresponding results in the primary dataset. Within-person stability over 10 years is reasonable for lipid, lipid-related, and polar metabolites, and varies by metabolite class. Additional studies are required to estimate within-person stability over 10 years of other metabolites groups.
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- 2021
6. Organ-Specific Metabolome Deciphering Cell Pathways to Cope with Mercury in Wild Fish (Golden Grey Mullet
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Giuseppe, De Marco, Fátima, Brandão, Patrícia, Pereira, Mário, Pacheco, and Tiziana, Cappello
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mercury pollution ,gills ,Chelon auratus ,NMR-based metabolomics ,fish metabolome ,liver ,polar metabolites ,Article - Abstract
Simple Summary Metabolomics is a powerful approach that is based on the identification in biological samples of metabolites, which production and levels may vary due to factors intrinsic to the environment and the organism. For a correct data interpretation, it is, therefore, necessary to first evaluate the metabolome of the tissue/organ under investigation when it is exposed to no stressor. In this study, the complete set of metabolites of liver and gills of wild golden grey mullet (Chelon auratus) that were collected from a reference area was compared by using metabolomics, which was able to reveal metabolites that are commonly present in both organs but with different levels to be attributed to organ-specific functions. The same metabolomic approach was applied also to study the metabolite changes that were induced in mullet gills and liver after environmental exposure to mercury (Hg), and a variety of organ-specific metabolic disturbances were observed. The findings from this study validate the use of metabolomics in ecotoxicological studies to assess organ-specific functions and the cytotoxicity mechanisms of Hg in fish. Abstract Metabolomics is a powerful approach in evaluating the health status of organisms in ecotoxicological studies. However, metabolomics data reflect metabolic variations that are attributable to factors intrinsic to the environment and organism, and it is thus crucial to accurately evaluate the metabolome of the tissue/organ examined when it is exposed to no stressor. The metabolomes of the liver and gills of wild golden grey mullet (Chelon auratus) from a reference area were analyzed and compared by proton nuclear magnetic resonance (1H NMR)-based metabolomics. Both organs were characterized by amino acids, carbohydrates, osmolytes, nucleosides and their derivatives, and miscellaneous metabolites. However, similarities and differences were revealed in their metabolite profile and related to organ-specific functions. Taurine was predominant in both organs due to its involvement in osmoregulation in gills, and detoxification and antioxidant protective processes in liver. Environmental exposure to mercury (Hg) triggered multiple and often differential metabolic alterations in fish organs. Disturbances in ion-osmoregulatory processes were highlighted in the gills, whereas differential impairments between fish organs were pointed out in energy-producing metabolic pathways, protein catabolism, membrane stabilization processes, and antioxidant defense system, reflecting the induction of organ-specific adaptive and defensive strategies. Overall, a strict correlation between metabolites and organ-specific functions of fish gills and liver were discerned in this study, as well as organ-specific cytotoxicity mechanisms of Hg in fish.
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- 2021
7. Metabolomic and biochemical analysis of mesocarp tissues from table grape berries with contrasting firmness reveals cell wall modifications associated to harvest and cold storage
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Iván Balic, Patricio Olmedo, Baltasar Zepeda, Bárbara Rojas, Troy Ejsmentewicz, Miriam Barros, Daniel Aguayo, Adrián A. Moreno, Romina Pedreschi, Claudio Meneses, and Reinaldo Campos-Vargas
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Polar metabolites ,Horticulture & Product Physiology ,General Medicine ,PE&RC ,Pectin ,Analytical Chemistry ,Cell Wall ,Fruit ,Vitis vinifera ,Metabolomics ,Vitis ,Calcium ,Texture ,Tuinbouw & Productfysiologie ,Food Science - Abstract
Tissue texture influences the grape berry consumers acceptance. We studied the biological differences between the inner and outer mesocarp tissues in hard and soft berries of table grapes cv NN107. Texture analysis revealed lower levels of firmness in the inner mesocarp as compared with the outer tissue. HPAEC-PAD analysis showed an increased abundance of cell wall monosaccharides in the inner mesocarp of harder berries at harvest. Immunohistochemical analysis displayed differences in homogalacturonan methylesterification and cell wall calcium between soft and hard berries. This last finding correlated with a differential abundance of calcium measured in the alcohol-insoluble residues (AIR) of the inner tissue of the hard berries. Analysis of abundance of polar metabolites suggested changes in cell wall carbon supply precursors, providing new clues in the identification of the biochemical factors that define the texture of the mesocarp of grape berries.
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- 2022
8. A Sample Preparation Method for the Simultaneous Profiling of Signaling Lipids and Polar Metabolites in Small Quantities of Muscle Tissues from a Mouse Model for Sarcopenia
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Yupeng He, Marlien van Mever, Wei Yang, Luojiao Huang, Rawi Ramautar, Yvonne Rijksen, Wilbert P. Vermeij, Jan H. J. Hoeijmakers, Amy C. Harms, Peter W. Lindenburg, Thomas Hankemeier, and Molecular Genetics
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SDG 3 - Good Health and Well-being ,Endocrinology, Diabetes and Metabolism ,Molecular Biology ,Biochemistry ,metabolomics extraction ,signaling lipids ,polar metabolites ,muscle tissue ,muscle ageing and sarcopenia - Abstract
The metabolic profiling of a wide range of chemical classes relevant to understanding sarcopenia under conditions in which sample availability is limited, e.g., from mouse models, small muscles, or muscle biopsies, is desired. Several existing metabolomics platforms that include diverse classes of signaling lipids, energy metabolites, and amino acids and amines would be informative for suspected biochemical pathways involved in sarcopenia. The sample limitation requires an optimized sample preparation method with minimal losses during isolation and handling and maximal accuracy and reproducibility. Here, two developed sample preparation methods, BuOH-MTBE-Water (BMW) and BuOH-MTBE-More-Water (BMMW), were evaluated and compared with previously reported methods, Bligh-Dyer (BD) and BuOH-MTBE-Citrate (BMC), for their suitability for these classes. The most optimal extraction was found to be the BMMW method, with the highest extraction recovery of 63% for the signaling lipids and 81% for polar metabolites, and an acceptable matrix effect (close to 1.0) for all metabolites of interest. The BMMW method was applied on muscle tissues as small as 5 mg (dry weight) from the well-characterized, prematurely aging, DNA repair-deficient Ercc1∆/− mouse mutant exhibiting multiple–morbidities, including sarcopenia. We successfully detected 109 lipids and 62 polar targeted metabolites. We further investigated whether fast muscle tissue isolation is necessary for mouse sarcopenia studies. A muscle isolation procedure involving 15 min at room temperature revealed a subset of metabolites to be unstable; hence, fast sample isolation is critical, especially for more oxidative muscles. Therefore, BMMW and fast muscle tissue isolation are recommended for future sarcopenia studies. This research provides a sensitive sample preparation method for the simultaneous extraction of non-polar and polar metabolites from limited amounts of muscle tissue, supplies a stable mouse muscle tissue collection method, and methodologically supports future metabolomic mechanistic studies of sarcopenia.
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- 2022
9. An exploratory approach for an oriented development of an untargeted hydrophilic interaction liquid chromatography-mass spectrometry platform for polar metabolites in biological matrices
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Katyeny Manuela Da Silva, Matthias Cuykx, Adrian Covaci, Elias Iturrospe, Joost Boeckmans, Alexander L.N. van Nuijs, Tamara Vanhaecke, Begoña Talavera Andújar, Faculty of Medicine and Pharmacy, Experimental in vitro toxicology and dermato-cosmetology, Pharmaceutical and Pharmacological Sciences, Connexin Signalling Research Group, and Biology
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Polar metabolites ,Method optimization ,Health, Toxicology and Mutagenesis ,Dermatology ,010402 general chemistry ,Mass spectrometry ,01 natural sciences ,Biochemistry ,Mass Spectrometry ,Analytical Chemistry ,Biological pathway ,Liquid chromatography-high resolution mass spectrometry ,Plasma ,Metabolomics ,Metabolome ,Humans ,Biology ,chemistry.chemical_classification ,Chromatography ,Chemistry ,Hydrophilic interaction chromatography ,Liver cell ,010401 analytical chemistry ,Organic Chemistry ,Hydrophilic Interaction Chromatography ,General Medicine ,0104 chemical sciences ,Amino acid ,Body Fluids ,Metabolic pathway ,Human medicine ,Hydrophobic and Hydrophilic Interactions ,Chromatography, Liquid - Abstract
The analysis of polar metabolites based on liquid chromatography-mass spectrometry (LC-MS) methods should take into consideration the complexity of interactions in LC columns to be able to cover a broad range of metabolites of key biological pathways. Therefore, in this study, different chromatographic columns were tested for polar metabolites including reversed-phase and hydrophilic interaction liquid chromatography (HILIC) columns. Based on a column screening, two new generations of zwitterionic HILIC columns were selected for further evaluation. A tree-based method optimization was applied to investigate the chromatographic factors affecting the retention mechanisms of polar metabolites with zwitterionic stationary phases. The results were evaluated based on a scoring system which was applied for more than 80 polar metabolites with a high coverage of key human metabolic pathways. The final optimized methods showed high complementarity to analyze a wide range of metabolic classes including amino acids, small peptides, sugars, amino sugars, phosphorylated sugars, organic acids, nucleobases, nucleosides, nucleotides and acylcarnitines. Optimized methods were applied to analyze different biological matrices, including human urine, plasma and liver cell extracts using an untargeted approach. The number of high-quality features (< 30% median relative standard deviation) ranged from 3,755 for urine to 5,402 for the intracellular metabolome of liver cells, showing the potential of the methods for untargeted purposes.
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- 2020
10. Human and Aquatic Toxicity Potential of Petroleum Biodegradation Metabolite Mixtures in Groundwater from Fuel Release Sites
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Kirk T. O'Reilly, Natasha Sihota, Rachel E. Mohler, Lauren Kristofco, Catalina Espino Devine, Dawn A. Zemo, Renae I. Magaw, Sungwoo Ahn, Timothy J. Patterson, and Asheesh K. Tiwary
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Polar metabolites ,Salinity ,Health, Toxicology and Mutagenesis ,Metabolite ,Cyprinidae ,Fresh Water ,010501 environmental sciences ,medicine.disease_cause ,Ecotoxicology ,01 natural sciences ,Aquatic toxicology ,03 medical and health sciences ,chemistry.chemical_compound ,Raphidocelis subcapitata ,Biotransformation ,Water Quality ,medicine ,Environmental Chemistry ,Animals ,Humans ,Hazard/Risk Assessment ,Oxygen‐containing organic compounds (OCOCs) ,Toxicity Tests, Chronic ,Groundwater ,030304 developmental biology ,0105 earth and related environmental sciences ,0303 health sciences ,Mixture toxicology ,biology ,Chemistry ,Ceriodaphnia dubia ,biology.organism_classification ,Cladocera ,Biodegradation, Environmental ,Petroleum ,Environmental chemistry ,Toxicity ,Environmental toxicology ,Genotoxicity ,Water Pollutants, Chemical - Abstract
The potential toxicity to human and aquatic receptors of petroleum fuel biodegradation metabolites (oxygen‐containing organic compounds [OCOCs]) in groundwater has been investigated as part of a multi‐year research program. Whole mixtures collected from locations upgradient and downgradient of multiple fuel release sites were tested using: 1) in vitro screening assays for human genotoxicity (the gamma‐H2AX assay) and estrogenic effects (estrogen receptor transcriptional activation assay), and 2) chronic aquatic toxicity tests in 3 species (Ceriodaphnia dubia, Raphidocelis subcapitata, and Pimephales promelas). In vitro screening assay results demonstrated that the mixtures did not cause genotoxic or estrogenic effects. No OCOC‐related aquatic toxicity was observed and when aquatic toxicity did occur, upgradient samples typically had the same response as samples downgradient of the release, indicating that background water quality was impacting the results. This information provides additional support for previous work that focused on the individual compounds and, taken together, indicates that OCOCs from petroleum degradation at fuel release sites are unlikely to cause toxicity to human or freshwater receptors at the concentrations present. Environ Toxicol Chem 2020;39:1634–1645. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
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- 2020
11. New supported liquid membrane for electromembrane extraction of polar basic endogenous metabolites
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Julie Schappler, Serge Rudaz, and Nicolas Drouin
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Male ,Spectrometry, Mass, Electrospray Ionization ,Polar metabolites ,Supported liquid membrane ,Bioanalysis ,Clinical Biochemistry ,Pharmaceutical Science ,02 engineering and technology ,01 natural sciences ,Capillary electrophoresis–mass spectrometry ,Analytical Chemistry ,Matrix (chemical analysis) ,Electromembrane extractionPa-EME ,Drug Discovery ,Humans ,Spectroscopy ,ddc:615 ,Chromatography ,Chemistry ,Chemical polarity ,010401 analytical chemistry ,Extraction (chemistry) ,Membranes, Artificial ,Electrochemical Techniques ,021001 nanoscience & nanotechnology ,Body Fluids ,0104 chemical sciences ,Partition coefficient ,Membrane ,Solvents ,Endogenous compound ,Polar ,Female ,Capillary electrophoresis-mass spectrometry ,0210 nano-technology - Abstract
Extraction of polar endogenous compounds remains an important issue in bioanalysis although different techniques have been evaluated. Among them, electromembrane extraction (EME) is a relevant approach but supported liquid membranes (SLMs) dedicated to polar molecules are still lacking. In this study 22 organic solvents were evaluated as SLMs on a set of 45 polar basic metabolites (log P from −5.7 to 1.5) from various biochemical families. To investigate a large variety of organic solvents, a parallel electromembrane extraction device was used and a constant current approach was applied to circumvent the heterogeneous conductivities of the different SLMs. Among the tested organic solvents, 2-nitrophenyl pentyl ether (NPPE) appeared the most efficient SLM with the extraction of a large variety of polar cationic metabolites, high extraction yields, and low extraction variabilities. The applied current and the composition of the acceptor and donor solutions were also evaluated and 300 μA per well and acetic acid 1% (v/v), both as acceptor and donor compartments, were the most efficient conditions. The new SLM and the optimized experimental parameters were successfully applied to the extraction of precipitated plasma samples. Although the extraction recovery decreased for most compounds in the biological matrix, process efficiency (PE) up to 90% and low extraction variability (RSD between 2 and 18%) were obtained for several very polar compounds such as choline or acetylcholine, emphasizing the potential of EME for polar compounds.
- Published
- 2018
12. A Quantitative HILIC-MS/MS Assay of the Metabolic Response of Huh-7 Cells Exposed to 2,3,7,8-Tetrachlorodibenzo
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Andrew D. Patterson, Jingwei Cai, Zhang Jintao, Yuan Tian, Qing Liu, Robert G. Nichols, Chao Yan, Yan Wang, and Philip B. Smith
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0301 basic medicine ,TCDD ,Arginine ,Endocrinology, Diabetes and Metabolism ,Metabolite ,lcsh:QR1-502 ,Tandem mass spectrometry ,Biochemistry ,lcsh:Microbiology ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,targeted metabolomics ,Proline ,Molecular Biology ,Alanine ,chemistry.chemical_classification ,Chromatography ,biology ,Hydrophilic interaction chromatography ,Aryl hydrocarbon receptor ,polar metabolites ,Amino acid ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,biology.protein ,HILIC–UHPLC–MS/MS - Abstract
A hydrophilic interaction liquid chromatography (HILIC)&ndash, ultra high-pressure liquid chromatography (UHPLC) coupled with tandem mass spectrometry (MS/MS) method was developed and applied to profile metabolite changes in human Huh-7 cells exposed to the potent aryl hydrocarbon receptor (AHR) ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Comparisons of sensitivity (limit of detection as low as 0.01 µ, M) and reproducibility (84% of compounds had an interday relative standard deviation (RSD) less than 10.0%, 83% of compounds had an intraday RSD less than 15.0%) were assessed for all the metabolites. The exposure of Huh-7 cells to the hepatotoxic carcinogen TCDD at low doses (1 nM and 10 nM for 4 h and 24 h, respectively) was reflected by the disturbance of amino acid metabolism, energy metabolism (glycolysis, TCA cycle), and nucleic acid metabolism. TCDD caused a significant decrease in amino acids such as serine, alanine, and proline while promoting an increase in arginine levels with 24 h treatment. Energy metabolism intermediates such as phosphoenolpyruvate and acetyl&ndash, CoA and nucleosides such as UMP, XMP, and CMP were also markedly decreased. These results support the application of HILIC&ndash, UHPLC&ndash, MS/MS for robust and reliable analysis of the cellular response to environmentally relevant toxicants at lower doses.
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- 2019
13. The toxicity of residual hydrocarbons and polar metabolites on Antarctic soil microbial diversity
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Pudasaini, Sarita
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Polar metabolites ,Residual hydrocarbons ,Bacteria ,Eukaryotes ,Antarctic soil ,Multi-species toxicity ,Bioremediation - Abstract
Hydrocarbon contamination in terrestrial Antarctica pose serious threats to the environment and the native species thriving. A legacy of fuel spills means that partially remediated soils are likely to contain fingerprints of unresolved complex mixture (UCM) which are comprised of residual hydrocarbons and polar metabolites. However, their presence in hydrocarbon contaminated soil and ecotoxicity information is largely unknown. This lack of knowledge raises concerns on their potential toxicity. Since terrestrial Antarctica is dominated by microorganisms, the main aim of this study was to explore toxicity of residual hydrocarbons and polar metabolites on endemic Antarctic soil microorganisms of Casey station. Here, we used a synthetic residual fuel mixture (RFM) comprising five compounds that are present in Special Antarctic Blend diesel fuel, a commonly used fuel at Casey station and found that the bacterial community was more sensitive to RFM than the eukaryotic community. Dose-response modelling showed that the total DNA community produce more sensitive and reliable effective concentration (EC) values indicating DNA a better option for future ecotoxicity evaluations within these soils. Of five indices evaluated, Simpson and Shannon were the most reliable indicators for the bacterial and eukaryotic communities, respectively. For characterization of polar metabolites, out of 12 aldehydes and two ketones tested in both clean and contaminated elutriates (that mimics leachate runoff), only nine aldehydes were detected, with the majority being biogenic. Acetaldehyde was present at the highest concentration (821 µg/l) in the partially remediated soil, inferring it to be a potential chemical indicator for hydrocarbon degradation. Three major aldehydes (acetaldehyde, octanal, and undecanal) detected in the contaminated soil elutriates were selected for toxicity assessments using the single species toxicity assay Microtox (Aliivibrio fischeri) but were found below toxic level. The cumulative effect of water-soluble toxicants present in the contaminated soil elutriates were found toxic to Aliivibrio fischeri and site-specific bacterial communities. A higher proportion of bacterial genera (95-135) responded significantly (stimulated or inhibited) to contaminated soil elutriates. Our study found that both RFM and polar metabolites to have toxic effect on microbial communities, thus present a serious risk to the sensitive Antarctic environment and its microbial communities.
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- 2019
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14. Optimization of 1D
- Author
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Hyun Cheol, Kim, Yoon-Joo, Ko, Minsu, Kim, Juhui, Choe, Hae In, Yong, and Cheorun, Jo
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reconstitution buffer ,meat ,extraction solution ,qNMR ,polar metabolites ,Article - Abstract
The objective of this study was to establish an optimized 1D 1H quantitative nuclear magnetic resonance (qNMR) analytical method for analyzing polar metabolites in meat. Three extraction solutions [0.6 M perchloric acid, 10 mM phosphate buffer, water/methanol (1:1)], three reconstitution buffers [20 mM 3-morpholinopropane-1-sulfonic acid, 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, phosphate buffer], and two pulse programs (zg30, noesypr1d) were evaluated. Extraction with 0.6 M perchloric acid and 20 mM phosphate resulted in a stable baseline and no additional overlap for quantifying polar metabolites in chicken breast. In qNMR analysis, zg30 pulse program (without water-suppression) showed smaller relative standard deviation (RSD) and faster running time than noesypr1d (water-suppression). High-performance liquid chromatography was compared with qNMR analyses to validate accuracy. The zg30 pulse program showed good accuracy and lower RSD. The optimized qNMR method was able to apply for beef and pork samples. Thus, an optimized 1D 1H qNMR method for meat metabolomics was established.
- Published
- 2018
15. Simultaneous Quantitative Analysis of Metabolites Using Ion-Pair Liquid Chromatography−Electrospray Ionization Mass Spectrometry
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S. Jespersen, Thomas Hankemeier, Mariët J. van der Werf, Richard C. Bas, Elwin Verheij, Leon Coulier, and TNO Kwaliteit van Leven TNO Voeding
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Polar metabolites ,Electrospray ,Unclassified drug ,Ion chromatography ,Alkalinity ,Analytical chemistry ,Metabolite ,Quantitative assay ,Analytical Chemistry ,Sugar nucleotide ,chemistry.chemical_compound ,Metabolites ,Chemical analysis ,Electrospray mass spectrometry ,Amines ,Sampling ,Nucleotides ,Chemistry ,PH effects ,Esters ,Hydrogen-Ion Concentration ,Reproducibility ,Ammonium acetate ,Hexylamine ,Bisphosphonic acid derivative ,Nucleotide ,Biotechnology ,Bacillus subtilis ,Optimization ,Spectrometry, Mass, Electrospray Ionization ,Electrospray ionization ,Liquid chromatography ,Ion pair liquid chromatography ,Sugar bisphosphonate ,Acyl coenzyme A ,Mass spectrometry ,Electrospray ionization mass spectrometry ,Analytic method ,Escherichia coli ,Metabolomics ,Validation process ,Coenzyme A ,Biology ,Detection limit ,Chemical ionization ,Chromatography ,Ion pair chromatography ,Methanol ,Acidity ,Water ,Reproducibility of Results ,Energy charge ,Nonhuman ,Energy expenditure ,Sugar Phosphates ,Quantitative analysis (chemistry) ,Process optimization ,Chromatography, Liquid - Abstract
We have developed an analytical method, consisting of ion-pair liquid chromatography coupled to electrospray ionization mass spectrometry (IP-LC-ESI-MS), for the simultaneous quantitative analysis of several key classes of polar metabolites, like nucleotides, coenzyme A esters, sugar nucleotides, and sugar bisphosphates. The use of the ion-pair agent hexylamine and optimization of the pH of the mobile phases were critical parameters in obtaining good retention and peak shapes of many of the above-mentioned polar and acidic metabolites that are impossible to analyze using standard reversed-phase LC/MS. Optimum conditions were found when using a gradient from 5 mM hexylamine in water (pH 6.3) to 90% methanol/10% 10 mM ammonium acetate (pH 8.5). The IP-LC-ESI-MS method was extensively validated by determining the linearity (R2 > 0.995), sensitivity (limit of detection 0.1-1 ng), repeatability, and reproducibility (relative standard deviation
- Published
- 2006
16. Evaluation of extraction protocols for simultaneous polar and non-polar yeast metabolite analysis using multivariate projection methods
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Raymond J. Turner, Vanina Zaremberg, Nicolas P. Tambellini, and Aalim M. Weljie
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Analyte ,Endocrinology, Diabetes and Metabolism ,Metabolite ,OPLS ,lcsh:QR1-502 ,01 natural sciences ,Biochemistry ,chemoinformatics ,lcsh:Microbiology ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Metabolomics ,Methoxyamine ,chloroform/methanol/water partitioning ,Molecular Biology ,fatty acid methyl esters ,030304 developmental biology ,metabolite extraction ,chemistry.chemical_classification ,GC-MS ,polar metabolites ,PCA ,0303 health sciences ,Chromatography ,Chemistry ,010401 analytical chemistry ,Extraction (chemistry) ,Fatty acid ,0104 chemical sciences ,Polar ,Gas chromatography–mass spectrometry - Abstract
Metabolomic and lipidomic approaches aim to measure metabolites or lipids in the cell. Metabolite extraction is a key step in obtaining useful and reliable data for successful metabolite studies. Significant efforts have been made to identify the optimal extraction protocol for various platforms and biological systems, for both polar and non-polar metabolites. Here we report an approach utilizing chemoinformatics for systematic comparison of protocols to extract both from a single sample of the model yeast organism Saccharomyces cerevisiae. Three chloroform/methanol/water partitioning based extraction protocols found in literature were evaluated for their effectiveness at reproducibly extracting both polar and non-polar metabolites. Fatty acid methyl esters and methoxyamine/trimethylsilyl derivatized aqueous compounds were analyzed by gas chromatography mass spectrometry to evaluate non-polar or polar metabolite analysis. The comparative breadth and amount of recovered metabolites was evaluated using multivariate projection methods. This approach identified an optimal protocol consisting of 64 identified polar metabolites from 105 ion hits and 12 fatty acids recovered, and will potentially attenuate the error and variation associated with combining metabolite profiles from different samples for untargeted analysis with both polar and non-polar analytes. It also confirmed the value of using multivariate projection methods to compare established extraction protocols.
- Published
- 2013
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