RESUMEN
The tricarboxylic acid (TCA) cycle is one of the most important metabolic pathway for cellular respiration in aerobic organisms. It provides and collects intermediates for many other interconnecting pathways and acts as a hub connecting metabolism of carbohydrates, fatty acids, and amino acids. Alteration in intracellular levels of its intermediates has been linked with a wide range of illnesses ranging from cancer to cellular necrosis or liver cirrhosis. Therefore, there exists an intrinsic interest in monitoring such metabolites. Our goal in this study was to evaluate whether, at least the most volatile metabolites of the TCA cycle, could be detected in breath in vivo and in real time. We used secondary electrospray ionization coupled with high-resolution mass spectrometry (SESI-HRMS) to conduct this targeted analysis. We enrolled six healthy individuals who provided full exhalations into the SESI-HRMS system at different times during 3 days. For the first time, we observed exhaled compounds that appertain to the TCA cycle: fumaric, succinic, malic, keto-glutaric, oxaloacetic, and aconitic acids. We found high intraindividual variability and a significant overall difference between morning and afternoon levels for malic acid, oxaloacetic acid, and aconitic acid, supporting previous studies suggesting circadian fluctuations of these metabolites in humans. This study provides first evidence that TCA cycle could conveniently be monitored in breath, opening new opportunities to study in vivo this important metabolic pathway.
Asunto(s)
Pruebas Respiratorias/métodos , Ciclo del Ácido Cítrico , Espectrometría de Masa por Ionización de Electrospray/métodos , Ácidos Tricarboxílicos/análisis , Adulto , Pruebas Respiratorias/instrumentación , Diseño de Equipo , Espiración , Femenino , Humanos , Masculino , Espectrometría de Masa por Ionización de Electrospray/instrumentación , Espectrometría de Masas en Tándem/instrumentación , Espectrometría de Masas en Tándem/métodos , Ácidos Tricarboxílicos/metabolismoRESUMEN
Tumor markers are most popularly used in diagnosis of various cancers clinically. However, the confounding factors of individual background diversities, such as genetics, food preferences, living styles, physical exercises, etc., greatly challenge the identification of tumor markers. Study of the metabolic impact of inoculated tumors on model animals can facilitate the identification of metabolomic markers relevant to tumor insult. In this study, serum metabolites from nude mice (n = 14) inoculated with human H460 cells (human nonsmall cell lung carcinoma) were profiled using gas chromatography time-of-flight mass spectrometry. The mice with inoculated tumors showed an obviously different metabolic pattern from the control; identification of the discriminatory metabolites suggested the metabolic perturbation of free fatty acids, amino acids, glycolysis and tricarboxylic acid (TCA) cycle turnover. The significantly decreased TCA intermediates, free fatty acids, 3-hydroxybutyric acid and fluctuating amino acids (t-test, p < 0.05) in serum of tumor-bearing mice characterized the metabolic impact of local inoculated H460 tumor cells on the whole system. This indicates that they are candidate metabolomic markers for translational study of lung cancer, clinically.
Asunto(s)
Metaboloma/fisiología , Metabolómica/métodos , Neoplasias Experimentales/metabolismo , Aminoácidos/análisis , Aminoácidos/metabolismo , Animales , Línea Celular Tumoral , Ácidos Grasos/análisis , Ácidos Grasos/metabolismo , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Análisis de Componente Principal , Ácidos Tricarboxílicos/análisis , Ácidos Tricarboxílicos/metabolismoRESUMEN
The tricarboxylic acid (TCA) cycle is an interface among glycolysis, lipid metabolism, and amino acid metabolism. Increasing interest in cancer metabolism has created a demand for rapid and sensitive methods for quantifying the TCA cycle intermediates and related organic acids. We have developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify the TCA cycle intermediates in a 96-well format after O-benzylhydroxylamine (O-BHA) derivatization under aqueous conditions. This method was validated for quantitation of all common TCA cycle intermediates with good sensitivity, including α-ketoglutarate, malate, fumarate, succinate, 2-hydroxyglutarate, citrate, oxaloacetate, pyruvate, isocitrate, and lactate using a 8-min run time in cancer cells and tissues. The method was used to detect and quantify changes in metabolite levels in cancer cells and tumor tissues treated with a pharmacological inhibitor of nicotinamide phosphoribosyl transferase (NAMPT). This method is rapid, sensitive, and reproducible, and it can be used to assess metabolic changes in cancer cells and tumor samples.
Asunto(s)
Ciclo del Ácido Cítrico , Hidroxilaminas/química , Espectrometría de Masas/métodos , Neoplasias/metabolismo , Ácidos Tricarboxílicos/metabolismo , Línea Celular Tumoral , Cromatografía Liquida , Citocinas/antagonistas & inhibidores , Citocinas/metabolismo , Inhibidores Enzimáticos/farmacología , Humanos , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/metabolismo , Neoplasias/patología , Nicotinamida Fosforribosiltransferasa/antagonistas & inhibidores , Nicotinamida Fosforribosiltransferasa/metabolismo , Ácidos Tricarboxílicos/análisis , Ácidos Tricarboxílicos/químicaRESUMEN
The tricarboxylic acid (TCA) cycle is involved in the complete oxidation of organic acids to carbon dioxide in aerobic cells. It not only uses the acetyl-CoA derived from glycolysis but also uses breakdown products of proteins, fatty acids, and nucleic acids. Therefore, the TCA cycle involves numerous carbon fluxes through central metabolism to produce reductant power and transfer the generated electrons to the aerobic electron transport system where energy is formed by oxidative phosphorylation. Although the TCA cycle plays a crucial role in aerobic organisms and tissues, the lack of direct isotopic labeling information in its intermediates (organic acids) makes the quantification of its metabolic fluxes rather approximate. This is the major technical gap that this study intended to fill. In this work, we established and validated liquid and gas chromatography-mass spectrometry methods to determine (13)C labeling in organic acids involved in the TCA cycle using scheduled multiple reaction monitoring and single ion monitoring modes, respectively. Labeled samples were generated using maize embryos cultured with [(13)C]glucose or [(13)C]glutamine. Once steady-state labeling was reached, (13)C-labeled organic acids were extracted and purified. When applying our mass spectrometric methods to those extracts, mass isotopomer abundances of seven major organic acids were successfully determined.
Asunto(s)
Isótopos de Carbono/química , Ciclo del Ácido Cítrico , Cromatografía de Gases y Espectrometría de Masas/métodos , Marcaje Isotópico/métodos , Ácidos Tricarboxílicos/análisis , Glucosa/metabolismo , Glutamina/metabolismo , Reproducibilidad de los Resultados , Semillas/química , Semillas/fisiología , Técnicas de Cultivo de Tejidos , Zea mays/metabolismoRESUMEN
Porous Cu-BTC material was synthesized by the solvothermal method. Powder X-ray diffraction (PXRD) was used to test the phase purity of the synthesized material and investigate its structural stability under the influence of flue gas components. The thermal stability of the material was determined through thermal gravimetric (TG) analysis. Scanning electron microscopy (SEM) was employed to study the microstructure of the material. Cu-BTC was demonstrated not only to have high CO2 adsorption capacity but also good selectivity of CO2 over N2 by means of packed bed tests. The adsorption capacity of Cu-BTC for CO2 was about 69 mL/g at 22 degrees C. The influence of the main flue gas components on the CO2 capacity of the material were discussed as well.
Asunto(s)
Contaminantes Atmosféricos/análisis , Derivados del Benceno/análisis , Derivados del Benceno/síntesis química , Dióxido de Carbono/análisis , Ácidos Tricarboxílicos/análisis , Ácidos Tricarboxílicos/síntesis química , Adsorción , Cromatografía de Gases , Humedad , Microscopía Electrónica de Rastreo , Nitrógeno/análisis , Polvos , Termogravimetría , Difracción de Rayos XRESUMEN
The quantification of trimesic acid, a constitutive organic linker from the biodegradable porous iron(III) trimesate MIL-100(Fe) (MIL stands for Materials from Institut Lavoisier), has been performed in different biological complex media (liver, spleen and urine) using a liquid-liquid extraction procedure. A recovery exceeding 92 wt% was achieved from rat tissues and urine spiked with trimesic acid. After extraction, the determination of the trimesic acid concentration was realised by using a simple and accurate high-performance liquid chromatography (HPLC) method using photodiode-array detection (PDA) and aminosalicylic acid, as internal standard. Linearity of this method was kept from 0.01 to 100mg of trimesic acid per liter of urine and from 0.05 to 5.00 wt% of trimesic acid per tissue weight. The limit of detection of the method was 0.01 µg per injection. This method was finally applied to analyze and quantify the amount of trimesic acid in rat urine and tissue samples at the different stages of degradation of MIL-100(Fe).
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Hígado/química , Bazo/química , Ácidos Tricarboxílicos/análisis , Orina/química , Animales , Cromatografía Líquida de Alta Presión/instrumentación , Femenino , Ratas , Ratas Wistar , Ácidos Tricarboxílicos/orinaRESUMEN
BACKGROUND: Citric, malic, tartaric, oxalic, ascorbic, aconitic and fumaric acids, and sucrose, fructose and glucose, were determined in ten potato cultivars (Azucena negra, Bonita, Bonita negra, Borralla, Colorada de baga, Mora, Negra, Palmera lagarteada, Peluca blanca and Terrenta) from the Canary Islands in order to differentiate them. The influence of the production zone was also considered. RESULTS: Citric acid and sucrose were the most abundant organic acid and sugar, respectively. There were significant differences in all the analysed sugars and organic acids between the potato cultivars, except for oxalic and malic acids. Differences associated with the production zone were found in the contents of organic acids for some cultivars. Linear discriminant analysis is a useful tool to differentiate the potato samples according to the cultivar and the zone of production. CONCLUSIONS: Seven organic acids (citric, oxalic, malic, tartaric, aconitic, fumaric, and ascorbic acids) were identified and quantified in potatoes. The main sugars (sucrose, glucose and fructose) were also determined. The genetic information of the cultivars determines marked differences in sugars and organic acid contents among the analysed potato cultivars. The soil and climatic conditions affect the organic acid and sugar contents of some potato cultivars.
Asunto(s)
Ácido Ascórbico/análisis , Ácidos Dicarboxílicos/análisis , Sacarosa en la Dieta/análisis , Hexosas/análisis , Tubérculos de la Planta/química , Solanum tuberosum/química , Ácidos Tricarboxílicos/análisis , Islas del Atlántico , Cromatografía Líquida de Alta Presión , Fructosa/análisis , Glucosa/análisis , Concentración de Iones de Hidrógeno , Suelo , España , Especificidad de la Especie , Estadística como AsuntoRESUMEN
The migration of trimellitic acid and its esters from epoxy anhydride coatings was determined in simulants as well as in canned foods. The most appropriate simulant was a combination of EC simulants B and C: 2% acetic acid/10% ethanol in water. The average migration into food was 900 microg kg(-1). This far exceeds the 50 microg kg(-1) for which the safety of trimellitic acid and its anhydride is ensured and the Swiss legal limit (QM(T) of 5 mg kg(-1) coating). Furthermore, much trimellitic acid migrated as (unidentified) esters, i.e. toxicological testing of free trimellitic acid is inadequate for the material that in reality migrates.
Asunto(s)
Compuestos Epoxi , Contaminación de Alimentos/análisis , Embalaje de Alimentos/métodos , Ácidos Tricarboxílicos/análisis , Cromatografía de Gases/métodos , Cromatografía Líquida de Alta Presión/métodos , Ésteres/análisis , Malus/química , Metilación , Pisum sativum/química , Estándares de Referencia , Ácidos Tricarboxílicos/químicaRESUMEN
Fumonisins are a structurally related group of mycotoxins, characterized by a 19-20 carbon aminopolyhydroxy-alkyl chain which is diesterified with propane-1,2,3-tricarboxylic acid (tricarballylic acid). These mycotoxins are commonly found in corn and corn-based food products and have been linked to a variety of animal toxicities. The widespread prevalence of fumonisins and the toxicity associated with ingestion has resulted in a number of analytical methods for determining the amount of fumonisins present in foods. Among the most common of these methods are liquid chromatographic (LC) separation with fluorescence detection, enzyme-linked immunosorbent assay (ELISA) and LC/mass spectrometry. LC and ELISA give quantitative results while LC/MS provide quantitative analysis as well as confirmation of identity of the fumonisins.
Asunto(s)
Micotoxinas/análisis , Cromatografía Líquida de Alta Presión , Ensayo de Inmunoadsorción Enzimática , Fusarium/química , Estándares de Referencia , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray , Ácidos Tricarboxílicos/análisis , Zea mays/químicaRESUMEN
Three unknown di- and tricarboxylic acids were characterized in the fine size fraction of aerosols which were collected during the wet season in the Amazon basin (Rondonia, Brazil). For the structural characterization of the methyl esters of these unknown compounds, mass spectrometry with electron ionization (EI) and tandem mass spectral techniques combined with gas chromatographic (GC) separation were employed. Fragment and parent ion spectra were recorded during elution of the GC peaks by linked scanning of the B and E sectors in combination with high-energy collision-induced dissociation. The fragmentation patterns of significant ions in the first-order EI spectra were also obtained for nonanedioic acid, which was examined as a model compound. The compounds were tentatively identified as 4-acetyloxyheptanedioic acid and cis and trans isomers of 5-hexene-1,1,6-tricarboxylic acid. Since there were indications of biomass burning during the aerosol sampling the di- and tricarboxylic acids characterized in the present work could be markers for biomass burning. Furthermore, the characterization of di- and tricarboxylic acids in the fine size fraction of atmospheric aerosols may be important for assessing the effects of organic aerosols in cloud formation.
Asunto(s)
Contaminantes Atmosféricos/análisis , Atmósfera/análisis , Ácidos Dicarboxílicos/análisis , Ácidos Tricarboxílicos/análisis , Clima Tropical , Aerosoles/química , Cromatografía de Gases y Espectrometría de Masas , Lluvia , Estaciones del Año , Espectrometría de Masa por Ionización de Electrospray/métodosRESUMEN
The vnf-encoded nitrogenase from Azotobacter vinelandii contains an iron-vanadium cofactor (FeV-co) in its active site. Little is known about the synthesis pathway of FeV-co, other than that some of the gene products required are also involved in the synthesis of the iron-molybdenum cofactor (FeMo-co) of the widely studied molybdenum-dinitrogenase. We have found that VnfX, the gene product of one of the genes contained in the vnf-regulon, accumulates iron and vanadium in a novel V-Fe cluster during synthesis of FeV-co. The electron paramagnetic resonance (EPR) and metal analyses of the V-Fe cluster accumulated on VnfX are consistent with a VFe7-8Sx precursor of FeV-co. The EPR spectrum of VnfX with the V-Fe cluster bound strongly resembles that of isolated FeV-co and a model VFe3S4 compound. The V-Fe cluster accumulating on VnfX does not contain homocitrate. No accumulation of V-Fe cluster on VnfX was observed in strains with deletions in genes known to be involved in the early steps of FeV-co synthesis, suggesting that it corresponds to a precursor of FeV-co. VnfX purified from a nifB strain incapable of FeV-co synthesis has a different electrophoretic mobility in native anoxic gels than does VnfX, which has the V-Fe cluster bound. NifB-co, the Fe and S precursor of FeMo-co (and presumably FeV-co), binds to VnfX purified from the nifB strain, producing a shift in its electrophoretic mobility on anoxic native gels. The data suggest that a precursor of FeV-co that contains vanadium and iron accumulates on VnfX, and thus, VnfX is involved in the synthesis of FeV-co.
Asunto(s)
Azotobacter vinelandii/enzimología , Metaloproteínas/biosíntesis , Nitrogenasa/química , Proteínas Bacterianas/química , Sitios de Unión , Espectroscopía de Resonancia por Spin del Electrón , Hierro/química , Metaloproteínas/química , Molibdeno/química , Molibdoferredoxina/biosíntesis , Molibdoferredoxina/química , Ácidos Tricarboxílicos/análisis , Vanadio/químicaRESUMEN
The alternative nitrogenase from a nifH mutant of the photosynthetic bacterium Rhodospirillum rubrum has been purified and characterized. The dinitrogenase protein (ANF1) contains three subunits in an apparent alpha2beta2gamma2 structure and contains Fe but no Mo or V. A factor capable of activating apo-dinitrogenase (lacking the FeMo cofactor) from Azotobacter vinelandii was extracted from the alternative dinitrogenase protein with N-methylformamide. The electron paramagnetic resonance (EPR) signal of the dinitrogenase protein is not characteristic of the EPR signals of molybdenum- or vanadium-containing dinitrogenases. The alternative dinitrogenase reductase (ANF2) was purified as an alpha2 dimer containing an Fe4S4 cluster and exhibited an EPR spectrum characteristic of dinitrogenase reductases. The enzyme complex reduces protons to H2 very well but reduces N2 to ammonium poorly. Acetylene is reduced to a mixture of ethylene and ethane.
Asunto(s)
Proteínas Bacterianas/química , Proteínas Hierro-Azufre/química , Isoenzimas/química , Nitrogenasa/química , Oxidorreductasas , Rhodospirillum rubrum/enzimología , Acetileno/metabolismo , Apoenzimas/química , Apoenzimas/metabolismo , Proteínas Bacterianas/metabolismo , Dinitrogenasa Reductasa/química , Dinitrogenasa Reductasa/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Electroforesis en Gel de Poliacrilamida , Etano/metabolismo , Hierro/análisis , Proteínas Hierro-Azufre/metabolismo , Isoenzimas/metabolismo , Metales/análisis , Mutación , Nitrogenasa/genética , Nitrogenasa/metabolismo , Oxidación-Reducción , Conformación Proteica , Compuestos de Amonio Cuaternario/metabolismo , Rhodospirillum rubrum/genética , Especificidad por Sustrato , Ácidos Tricarboxílicos/análisisRESUMEN
BACKGROUND: The development of myocardial protective strategies depends on a complete understanding of the pathophysiology of myocardial ischemia and reperfusion. This article reviews the rationale for inclusion of metabolic substrates in cardioplegic solutions on the basis of our current understanding of the underlying pathophysiologic pathways and speculates on the inclusion of future additives that await further investigation. METHODS: The pathophysiology of myocardial ischemia and reperfusion was evaluated from an extensive review of the pertinent literature. Experimental and clinical studies supporting the inclusion of metabolic substrates in clinical cardioplegic solutions were reviewed and summarized. Speculation on possible future additives to these formulas was made on the basis of encouraging, albeit preliminary, experimental data. RESULTS: Sound experimental and clinical evidence supports the inclusion of glucose, amino acids, calcium chelators, and oxygen as fundamental substrate additives to current cardioplegic solutions. Antioxidants, calcium-channel blockers, and tricarboxylic acid cycle intermediates may be of value. Adenosine, potassium-adenosine triphosphate channel modulators, and nitric oxide may join these lists after further research. CONCLUSIONS: Substrate enhancement of clinical cardioplegic solutions is based on physiologic principles that have been confirmed in the clinical setting. Further definition of the intricacies of myocardial ischemia and reperfusion promises to expand the current list of additives.
Asunto(s)
Soluciones Cardiopléjicas/análisis , Aminoácidos/análisis , Animales , Antioxidantes/análisis , Calcio/análisis , Bloqueadores de los Canales de Calcio/análisis , Soluciones Cardiopléjicas/metabolismo , Glucosa/análisis , Humanos , Isquemia Miocárdica/fisiopatología , Reperfusión Miocárdica , Miocardio/metabolismo , Especies Reactivas de Oxígeno/análisis , Ácidos Tricarboxílicos/análisisRESUMEN
It has been discovered that 1,2,3-benzenetricarboxylic acid and some 5-substituted derivatives are specific and competitive inhibitors of the mitochondrial tricarboxylate carrier. In order to characterize these acids a study was carried out using electron ionization (EI), chemical ionization (CI) and fast atom bombardment (FAB) mass spectrometry on the free acids, the potassium salt of 1,2,3-benzenetricarboxylic acid and the corresponding methyl esters. Unimolecular decomposition processes were also studied using B/E linked scan experiments in order to correlate peaks present in the FAB mass spectra of the above compounds. The FAB ionization method, as expected, was the most suitable for analysing the very polar and thermolabile acids and the only method which could be used for the analysis of the potassium salt. EI and CI turned out to be appropriate methods for analysing the less polar and thermostable trimethyl esters.
Asunto(s)
Derivados del Benceno/análisis , Proteínas Portadoras/antagonistas & inhibidores , Mitocondrias/química , Ácidos Tricarboxílicos/análisis , Derivados del Benceno/farmacología , Espectrometría de Masas/métodos , Espectrometría de Masa Bombardeada por Átomos Veloces , Ácidos Tricarboxílicos/farmacologíaRESUMEN
When apodinitrogenase (lacking FeMo-co) was activated with FeMo-co synthesized in vitro in the presence of 3H-labeled homocitrate, label was incorporated into dinitrogenase. The physical association of the label with FeMo-co was demonstrated by reisolation and purification of the cofactor from dinitrogenase. The presence of homocitrate in FeMo-co was established by NMR analysis of the organic acid extracted from dinitrogenase. Quantitation of homocitrate in dinitrogenase showed it to be present at a 1:1 ratio with molybdenum.
Asunto(s)
Ferredoxinas/biosíntesis , Klebsiella pneumoniae/enzimología , Molibdoferredoxina/biosíntesis , Nitrogenasa/biosíntesis , Ácidos Tricarboxílicos/metabolismo , Cromatografía DEAE-Celulosa , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Cinética , Espectroscopía de Resonancia Magnética , Molibdoferredoxina/aislamiento & purificación , Nitrogenasa/aislamiento & purificación , Ácidos Tricarboxílicos/análisisRESUMEN
A rapid reversed-phase high-performance liquid chromatographic method for the simultaneous determination of glutaric acid, phenylephrine, and benzyl alcohol in nasal spray has been developed. UV detection was utilized at 210 nm for the assay of glutaric acid and phenylephrine with an adjustment to 254 nm for the measurement of benzyl alcohol. Linearity and recovery data were obtained for each component in spiked placebo studies. An investigation of the retention mechanisms of the three components showed that phenylephrine was retained by ion-pairing with octanesulfonate anion while glutaric acid and benzyl alcohol partitioned as a suppressed ion and a neutral molecule, respectively. The method has been further extended to the reversed-phase separation of di- and tricarboxylic acids using a totally aqueous 0.0074 M phosphoric acid mobile phase. The retention of these acids was related to their octanol-water partition coefficients and structural variation.
Asunto(s)
Ácidos Alcanesulfónicos , Alcoholes Bencílicos/análisis , Compuestos de Bencilo/análisis , Ácidos Dicarboxílicos/análisis , Glutaratos/análisis , Fenilefrina/análisis , Ácidos Tricarboxílicos/análisis , Aerosoles , Alcanosulfonatos , Alcohol Bencilo , Cromatografía Líquida de Alta Presión , Indicadores y Reactivos , Ácidos Fosfóricos/análisis , Espectrofotometría UltravioletaRESUMEN
Methods are described for simultaneous quantitation of Krebs cycle and related acids by gas chromatography--mass spectrometry using deuterium-labelled acids and n-butyl-d9-esters of the organic acids as internal standards. Using sulphuric acid as esterification catalyst, only lactic, succinic, fumaric, malic, maleic and citric acids were found to be stable to hydrogen exchange and could be used as reference standards in the deuterated form. In contrast, pyruvic, oxalacetic, alpha-ketoglutaric and malonic acids were found to exchange their deuterium readily and could not be employed for this purpose. All the acids could be quantitated using n-butyl-d9-esters of reference organic acids as internal standards, following a separate preparation of the n-butyl derivatives of the unknown acids. The method is suitable for routine analysis of organic acids at the picogram level in perchloric acid extracts of tissues.
Asunto(s)
Ácidos Carboxílicos/análisis , Ácidos Tricarboxílicos/análisis , Adipatos/análisis , Animales , Ésteres/síntesis química , Cromatografía de Gases y Espectrometría de Masas/métodos , Hígado/metabolismo , RatasRESUMEN
During pregnancy the rat liver shows alterations in metabolism which apparently do not to occur in the non-pregnant animal. In our study, the following metabolite concentrations and enzyme activities in the liver of pregnant and non-pregnant rats were measured after fasting periods of 6 and 12 h: malate, fumarate, isocitrate, alpha-keto-glutarate, glutamate, malate-dehydrogenase, fumarate-hydratase, glutamate-dehydrogenase, isocitrate-dehydrogenase, aspartate-aminotransferase and after 12 h fast, the concentrations of acetyl coenzyme A and citrate. These results are discussed with regard to their possible importance for the maternal and fetal energy supply.
Asunto(s)
Ciclo del Ácido Cítrico , Hígado/metabolismo , Embarazo , Animales , Ayuno , Femenino , Hígado/análisis , Hígado/enzimología , Ratas , Ácidos Tricarboxílicos/análisisRESUMEN
Trimellitic anhydride has widespread usage in a variety of industrial processes. Inhalation of the airborne dust and fumes generated by these processes is the primary route of occupational exposure leading to a variety of respiratory maladies. Because of its extensive industrial usage and limited toxicological data, trimellitic anhydride was scheduled for toxicological evaluation. Analytical purity certification and chemical characterization of the chemical were required as prerequisites for such toxicological tests. Therefore, a sensitive and specific procedure for the analysis of trimellitic anhydride and its trimellitic acid impurity in admixture was developed. After methylation with diazomethane, the compounds were assayed by gas chromatography using flame ionization detection. Chromatography on 5% SP-2100 yielded two well separated peaks, which were used for quantitation of the parent compounds. Confirmational identity of trimellitic anhydride was determined by direct insertion probe mass spectrometry, while that of the anhydride methyl derivative and trimellitic acid trimethyl derivative was determined by gas chromatography-mass spectrometry.