Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 98
Filtrar
Más filtros

Banco de datos
País/Región como asunto
Tipo del documento
Intervalo de año de publicación
1.
Mass Spectrom Rev ; 42(5): 1625-1646, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-34870867

RESUMEN

Conservation of historic artifacts has been a multidisciplinary field from its very beginning. Traditionally, it has been and still is associated with the history of art. It applies knowledge from technical and basic sciences, adapting their solutions to its goals. At present, however, a new tendency is clearly emerging-scientific research is starting to play an increasingly important role not only as a service, but also by proposing new solutions both in the traditional conservation areas and in new areas of conservation activities. The above trend opens up new perspectives for the field of preservation of our heritage but may also create new threats. Therefore, the conservators' caution in introducing new technologies should always be justified; after all, they are responsible for the effects of any activities on the historic objects. This, quite selective review, discusses application of mass spectrometry techniques for the detection of various components that are important to the conservators of our heritage with particular focus on paintings. The text also contains some basic knowledge of technical details to introduce the methodology to a broader group of professionals.

2.
Mass Spectrom Rev ; 39(5-6): 453-470, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-31793697

RESUMEN

Expectations for continuous miniaturization in mass spectrometry are not declining for years. Portable instruments are highly welcome by the industry, science, space agencies, forensic laboratories, and many other units. All are striving for the small, cheap, and as good as possible instruments. This review describes the recent developments of miniature mass spectrometers and also provides selected applications where these devices are used. Upcoming perspectives of further development are also discussed. @ 2019 John Wiley & Sons Ltd. Mass Spec Rev.

3.
J Enzyme Inhib Med Chem ; 36(1): 183-187, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33401948

RESUMEN

We have applied a recently developed HPLC-MS enzymatic assay to investigate the cryptic peptides generated by the action of the insulin-degrading enzyme (IDE) on some neuropeptides (NPs) involved in the development of tolerance and dependence to opioids. Particularly, the tested NPs are generated from the NPFF precursor (pro-NPFF (A)): NPFF (FLFQPQRF) and NPAF (AGEGLSSPFWSLAAPQRF). The results show that IDE is able to cleave NPFF and NPAF, generating specific cryptic peptides. As IDE is also responsible for the processing of many other peptides in the brain (amyloid beta protein among the others), we have also performed competitive degradation assays using mixtures of insulin and the above mentioned NPs. Data show that insulin is able to slow down the degradation of both NPs tested, whereas, surprisingly, NPAF is able to accelerate insulin degradation, hinting IDE as the possible link responsible of the mutual influence between insulin and NPs metabolism.


Asunto(s)
Insulina/química , Insulisina/química , Antagonistas de Narcóticos/química , Neuropéptidos/química , Oligopéptidos/química , Fragmentos de Péptidos/química , Secuencia de Aminoácidos , Analgésicos Opioides/efectos adversos , Animales , Cromatografía Líquida de Alta Presión , Tolerancia a Medicamentos , Humanos , Insulina/metabolismo , Espectrometría de Masas , Neuropéptidos/metabolismo , Fragmentos de Péptidos/análisis , Proteolisis , Ratas , Proteínas Recombinantes/química , Soluciones
4.
Molecules ; 26(12)2021 Jun 09.
Artículo en Inglés | MEDLINE | ID: mdl-34207930

RESUMEN

Jacek Namiesnik, who died at the age of 69, was one of the most influential analytical chemists in Poland at the second half of the 20th century and the first two decades of the 21st century [...].


Asunto(s)
Técnicas de Química Analítica/métodos , Investigadores/historia , Toxicología/métodos , Vino/análisis , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Polonia
5.
Molecules ; 26(13)2021 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-34201982

RESUMEN

During the last three decades, a variety of different studies on bioactive peptides that are opioid receptor ligands, have been carried out, with regard to their isolation and identification, as well as their molecular functions in living organisms. Thus, in this review, we would like to summarize the present state-of-the art concerning hemorphins, methodological aspects of their identification, and their potential role as therapeutic agents. We have collected and discussed articles describing hemorphins, from their discovery up until now, thus presenting a very wide spectrum of their characteristic and applications. One of the major assets of the present paper is a combination of analytical and pharmacological aspects of peptides described by a team who participated in the initial research on hemorphins. This review is, in part, focused on the analysis of endogenous opioid peptides in biological samples using advanced techniques, description of the identification of synthetic/endogenous hemorphins, their involvement in pharmacology, learning, pain and other function. Finally, the part regarding hemorphin analogues and their synthesis, has been added.


Asunto(s)
Péptidos Opioides/metabolismo , Dolor/metabolismo , Fragmentos de Péptidos/metabolismo , Receptores Opioides/metabolismo , Animales , Humanos
6.
Molecules ; 25(4)2020 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-32069913

RESUMEN

This work presents the synthesis of the novel covalent inhibitor of cysteine proteases where epoxide has been replaced by the iodoacetyl functional group. The molecule, similar in action to E-64 and DCG-04, the commonly applied inhibitors, is additionally biotinylated and contains tyrosyl iodination sites. The Fmoc solid phase synthesis has been applied. Conjugation of iodoacetic acid with the peptide was optimized by testing different conjugation agents. The purity of the final product was verified by mass spectrometry and its bioactivity was tested by incubation with a model cysteine protease-staphopain C. Finally, it was shown that the synthesized inhibitor binds to the protein at the ratio of 1:1. More detailed analysis by means of tandem mass spectrometry proved that the inhibitor binds to the cysteine present in the active site of the enzyme.


Asunto(s)
Cisteína Endopeptidasas/química , Cisteína Endopeptidasas/síntesis química , Inhibidores de Cisteína Proteinasa/química , Inhibidores de Cisteína Proteinasa/síntesis química , Ácido Yodoacético/química , Biotinilación , Leucina/análogos & derivados , Leucina/química , Estructura Molecular , Técnicas de Síntesis en Fase Sólida/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
7.
Int J Mol Sci ; 20(18)2019 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-31509943

RESUMEN

Insulin-degrading enzyme (IDE) was applied to catalyze hydrolysis of Nociceptin/Orphanin 1-16 (OFQ/N) to show the involvement of the enzyme in degradation of neuropeptides engaged in pain transmission. Moreover, IDE degradative action towards insulin (Ins) was inhibited by the OFQ/N fragments, suggesting a possible regulatory mechanism in the central nervous system. It has been found that OFQ/N and Ins affect each other degradation by IDE, although in a different manner. Indeed, while the digestion of OFQ/N is significantly affected by the presence of Ins, the kinetic profile of the Ins hydrolysis is not affected by the presence of OFQ/N. However, the main hydrolytic fragments of OFQ/N produced by IDE exert inhibitory activity towards the IDE-mediated Ins degradation. Here, we present the results indicating that, besides Ins, IDE cleaves neuropeptides and their released fragments act as inhibitors of IDE activity toward Ins. Having in mind that IDE is present in the brain, which also contains Ins receptors, it cannot be excluded that this enzyme indirectly participates in neural communication of pain signals and that neuropeptides involved in pain transmission may contribute to the regulation of IDE activity. Finally, preliminary results on the metabolism of OFQ/N, carried out in the rat spinal cord homogenate in the presence of various inhibitors specific for different classes of proteases, show that OFQ/N proteolysis in rat spinal cord could be due, besides IDE, also to a cysteine protease not yet identified.


Asunto(s)
Insulina/metabolismo , Insulisina/metabolismo , Péptidos Opioides/metabolismo , Médula Espinal/metabolismo , Secuencia de Aminoácidos , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Cromatografía Liquida/métodos , Insulina/química , Insulisina/antagonistas & inhibidores , Espectrometría de Masas/métodos , Neuropéptidos/química , Neuropéptidos/metabolismo , Neuropéptidos/farmacología , Péptidos Opioides/química , Dolor/prevención & control , Dimensión del Dolor/métodos , Fragmentos de Péptidos/química , Fragmentos de Péptidos/farmacología , Ratas , Receptor de Insulina/metabolismo , Médula Espinal/efectos de los fármacos , Nociceptina
8.
J Proteome Res ; 17(6): 2174-2181, 2018 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-29703078

RESUMEN

Ever since the development of the process known as the systematic evolution of ligands by exponential enrichment (SELEX), aptamers have been widely used in a variety of studies, including the exploration of new diagnostic tools and the discovery of new treatment methods. Aptamers' ability to bind to proteins with high affinity and specificity, often compared to that of antibodies, enables the search for potential cancer biomarkers and helps us understand the mechanisms of carcinogenesis. The blind spot of those investigations is usually the difficulty in the selective extraction of targets attached to the aptamer. There are many studies describing the cell SELEX for the prime choice of aptamers toward living cancer cells or even whole tumors in the animal models. However, a dilemma arises when a large number of proteins are being identified as potential targets, which is often the case. In this article, we present a new analytical approach designed to selectively target proteins bound to aptamers. During studies, we have focused on the unambiguous identification of the molecular targets of aptamers characterized by high specificity to the prostate cancer cells. We have compared four assay approaches using electrophoretic and chromatographic methods for "fishing out" aptamer protein targets followed by mass spectrometry identification. We have established a new methodology, based on the fluorescent-tagged oligonucleotides commonly used for flow-cytometry experiments or as optic aptasensors, that allowed the detection of specific aptamer-protein interactions by mass spectrometry. The use of atto488-labeled aptamers for the tracking of the formation of specific aptamer-target complexes provides the possibility of studying putative protein counterparts without needing to apply enrichment techniques. Significantly, changes in the hydrophobic properties of atto488-labeled aptamer-protein complexes facilitate their separation by reverse-phase chromatography combined with fluorescence detection followed by mass-spectrometry-based protein identification. These comparative results of several methodological approaches confirmed the universal applicability of this method to studying aptamer-protein interactions with high sensitivity, showing superior properties compared with pull-down techniques.


Asunto(s)
Aptámeros de Péptidos/metabolismo , Fluoresceínas , Técnica SELEX de Producción de Aptámeros/métodos , Cromatografía , Electroforesis , Colorantes Fluorescentes , Humanos , Espectrometría de Masas , Métodos , Unión Proteica
9.
J Proteome Res ; 16(4): 1436-1444, 2017 04 07.
Artículo en Inglés | MEDLINE | ID: mdl-28244758

RESUMEN

After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. Most candidate biomarkers are focused on the protein component. Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. Glycosylation is one of the most common and functionally important PTMs. It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. Cancer-associated aberrant glycosylation has been identified in various types of cancer. Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls.


Asunto(s)
Biomarcadores de Tumor/sangre , Haptoglobinas/análisis , Subunidad alfa del Receptor del Factor Inhibidor de Leucemia/sangre , Proteínas de Microtúbulos/sangre , Proteínas Nucleares/sangre , Neoplasias Pancreáticas/sangre , Proteínas de Transporte Vesicular/sangre , Anciano , Epítopos/biosíntesis , Epítopos/química , Epítopos/genética , Femenino , Glicosilación , Interacciones Huésped-Patógeno/genética , Humanos , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Polisacáridos/biosíntesis , Polisacáridos/química , Polisacáridos/genética , Pliegue de Proteína , Procesamiento Proteico-Postraduccional/genética , Proteómica
10.
Mass Spectrom Rev ; 35(1): 22-34, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-25988731

RESUMEN

Plasma-based ambient ionization mass spectrometry techniques are gaining growing interest due to their specific features, such as the need for little or no sample preparation, its high analysis speed, and the ambient experimental conditions. Samples can be analyzed in gas, liquid, or solid forms. These techniques allow for a wide range of applications, like warfare agent detection, chemical reaction control, mass spectrometry imaging, polymer identification, and food safety monitoring, as well as applications in biomedical science, e.g., drug and pharmaceutical analysis, medical diagnostics, biochemical analysis, etc. Until now, the main drawback of plasma-based techniques is their quantitative aspect, but a lot of efforts have been done to improve this obstacle.


Asunto(s)
Técnicas de Química Analítica/métodos , Espectrometría de Masas/métodos , Diseño de Equipo , Espectrometría de Masas/instrumentación , Presión , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray/métodos
11.
Anal Bioanal Chem ; 409(13): 3393-3405, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28321502

RESUMEN

The application of molecularly imprinted polymers (MIPs) as molecular scavengers for ambient plasma ionization mass spectrometry has been reported for the first time. MIPs were synthesized using methacrylic acid as functional monomer; nicotine, propyphenazone, or methylparaben as templates; ethylene glycol dimethacrylate as a cross-linker; and 2,2'-azobisisobutyronitrile as polymerization initiator. To perform ambient plasma ionization experiments, a setup consisting of the heated crucible, a flowing atmospheric-pressure afterglow (FAPA) plasma ion source, and a quadrupole ion trap mass spectrometer has been used. The heated crucible with programmable temperature allows for desorption of the analytes from MIPs structure which results in their direct introduction into the ion stream. Limits of detection, linearity of the proposed analytical procedure, and selectivities have been determined for three analytes: nicotine, propyphenazone, and methylparaben. The analytes used were chosen from various classes of organic compounds to show the feasibility of the analytical procedure. The limits of detections (LODs) were 10 nM, 10, and 0.5 µM for nicotine, propyphenazone, and methylparaben, respectively. In comparison with the measurements performed for the non-imprinted polymers, the values of LODs were improved for at least one order of magnitude due to preconcentration of the sample and reduction of background noise, contributing to signal suppression. The described procedure has shown linearity in a broad range of concentrations. The overall time of single analysis is short and requires ca. 5 min. The developed technique was applied for the determination of nicotine, propyphenazone, and methylparaben in spiked real-life samples, with recovery of 94.6-98.4%. The proposed method is rapid, sensitive, and accurate which provides a new option for the detection of small organic compounds in various samples. Graphical abstract The experimental setup used for analysis.

12.
Molecules ; 22(11)2017 Oct 28.
Artículo en Inglés | MEDLINE | ID: mdl-29143778

RESUMEN

The 46th EuroCongress on Drug Synthesis and Analysis (ECDSA-2017) was arranged within the celebration of the 65th Anniversary of the Faculty of Pharmacy at Comenius University in Bratislava, Slovakia from 5-8 September 2017 to get together specialists in medicinal chemistry, organic synthesis, pharmaceutical analysis, screening of bioactive compounds, pharmacology and drug formulations; promote the exchange of scientific results, methods and ideas; and encourage cooperation between researchers from all over the world. The topic of the conference, "Drug Synthesis and Analysis," meant that the symposium welcomed all pharmacists and/or researchers (chemists, analysts, biologists) and students interested in scientific work dealing with investigations of biologically active compounds as potential drugs. The authors of this manuscript were plenary speakers and other participants of the symposium and members of their research teams. The following summary highlights the major points/topics of the meeting.


Asunto(s)
Composición de Medicamentos , Química Farmacéutica , Humanos , Colaboración Intersectorial , Farmacéuticos , Relación Estructura-Actividad Cuantitativa , Investigadores , Eslovaquia
13.
Anal Bioanal Chem ; 408(3): 815-23, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26563110

RESUMEN

A thin-layer chromatography-mass spectrometry (TLC-MS) setup for characterization of low molecular weight compounds separated on standard TLC plates has been constructed. This new approach successfully combines TLC separation, laser ablation, and ionization using flowing atmospheric pressure afterglow (FAPA) source. For the laser ablation, a low-priced 445-nm continuous-wave diode laser pointer, with a power of 1 W, was used. The combination of the simple, low-budget laser pointer and the FAPA ion source has made this experimental arrangement broadly available, also for small laboratories. The approach was successfully applied for the characterization of low molecular weight compounds separated on TLC plates, such as a mixture of pyrazole derivatives, alkaloids (nicotine and sparteine), and an extract from a drug tablet consisting of paracetamol, propyphenazone, and caffeine. The laser pointer used was capable of ablating organic compounds without the need of application of any additional substances (matrices, staining, etc.) on the TLC spots. The detection limit of the proposed method was estimated to be 35 ng/cm(2) of a pyrazole derivative.


Asunto(s)
Alcaloides/análisis , Cromatografía en Capa Delgada/métodos , Preparaciones Farmacéuticas/análisis , Pirazoles/análisis , Presión Atmosférica , Cromatografía en Capa Delgada/instrumentación , Terapia por Láser
14.
Analyst ; 140(17): 6138-44, 2015 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-26222687

RESUMEN

In this paper, a procedure for the preconcentration and transport of mixtures of acids, bases, and drug components to a mass spectrometer using magnetic scavengers is presented. Flowing atmospheric pressure afterglow mass spectrometry (FAPA-MS) was used as an analytical method for identification of the compounds by thermal desorption from the scavengers. The proposed procedure is fast and cheap, and does not involve time-consuming purification steps. The developed methodology can be applied for trapping harmful substances in minute quantities, to transport them to specialized, remotely located laboratories.


Asunto(s)
Espectrometría de Masas , Ácido Benzoico/análisis , Ácidos Cafeicos/análisis , Óxido Ferrosoférrico/química , Ácidos Ftálicos/análisis , Poliaminas/análisis
15.
Anal Chem ; 86(22): 11226-9, 2014 Nov 18.
Artículo en Inglés | MEDLINE | ID: mdl-25338295

RESUMEN

Storage and preconcentration of various molecules by molecular scavengers for thermal desorption and identification by mass spectrometry is presented. A dielectric barrier discharge ionization source combined with a heating element for the chemical characterization of amines and organic acids, initially trapped by molecular scavengers, is described. The developed technique can be applied for preconcentration of minute amounts of molecules in liquid and gaseous phases, as well as their transportation and thorough analysis. The method, operating at ambient pressure, can also be complementary to electron impact ionization, with no need for sample derivatization.

16.
Langmuir ; 30(17): 5015-25, 2014 May 06.
Artículo en Inglés | MEDLINE | ID: mdl-24697681

RESUMEN

Poly(ethylene glycol)s (PEGs) with different lengths were used as linkers during the preparation of peptide surfaces for protease detection. In the first approach, the PEG monolayers were prepared using a "grafting to" method on 3-aminopropyltrietoxysilane (APTES)-modified silicon wafers. Protected peptides with a fluorescent marker were synthesized by Fmoc solid phase synthesis. The protected peptide structures enabled their site-specific immobilization onto the PEG surfaces. Alternatively, the PEG-peptide surface was obtained by immobilizing a PEG-peptide conjugate directly onto the modified silicon wafer. The surfaces (composition, grafting density, hydrophilicity, and roughness) were characterized by time-of-flight-secondary ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS), contact angle (CA), and atomic force microscopy (AFM). Introducing the PEG linker between the peptide and surface increased their resistance toward nonspecific protein adsorption. The peptide surfaces were examined as analytical platforms to study the action of trypsin as a representative protease. The products of the enzymatic hydrolysis were analyzed by fluorescence spectroscopy, electrospray ionization-mass spectrometry (ESI-MS), and ToF-SIMS. Conclusions about the optimal length of the PEG linker for the analytical application of PEG-peptide surfaces were drawn. This work demonstrates an effective synthetic procedure to obtain PEG-peptide surfaces as attractive platforms for the development of peptide microarrays.


Asunto(s)
Bioensayo/métodos , Péptido Hidrolasas/metabolismo , Péptidos/química , Péptidos/metabolismo , Polietilenglicoles/química , Espectroscopía de Fotoelectrones , Espectrometría de Masa de Ion Secundario , Propiedades de Superficie
17.
Mol Cell Biochem ; 386(1-2): 189-98, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24104455

RESUMEN

Prostate cancer is one of the most common malignancies in men and is predicted to be the second leading cause of cancer-related deaths. After 6-18 months, hormone ablation treatment results in androgen-independent growth of cancer cells, metastasis and progression. The mechanism of androgen-independent growth of prostatic carcinoma cells is still unknown. Identification of factors that facilitate the transition from androgen-dependent to independent states is crucial in designing future diagnostics and medication strategies. To understand the biochemical meaning of hormone dependency deprivation, glycoproteins enriched profiles were compared between DU145 (hormone non-responding) and LNCaP (hormone responding) prostate cancer cells. These results allow for anticipation on the important role of glycosylation in malignant transformation. Both Tn antigen and complex antennary N-oligosaccharides were recognized. Their occurrence might be involved in the development and progression of tumor, and failure of hormone ablation therapy. Among identified proteins in androgen-sensitive cells nucleolin (P19338) was found that is widely described as apoptosis inhibitor, and also transporter of molecules from the membrane to the cytoplasm or nucleus. In addition, 14-3-3 protein family (P27348, P31946, P61981, P63104, P62258, Q04917, and P31947) was investigated across available databases as it forms stable complexes with glycoproteins. Our studies indicate that isoforms: sigma and eta were found in androgen-dependent prostate cancer cells, while other isoforms were present in androgen non-responding cells. 14-3-3 binding partners are involved in cancer pathogenesis. These findings may contribute to a better understanding of prostate cancer tumorigenesis and to a more efficient prognosis and individual therapy in a future. However, it still remains to be revealed how important those changes are for androgen dependency loss in prostate cancer patients carried out on clinically relevant populations.


Asunto(s)
Andrógenos/fisiología , Glicopéptidos/metabolismo , Neoplasias Hormono-Dependientes/fisiopatología , Proteoma , Western Blotting , Línea Celular Tumoral , Transformación Celular Neoplásica , Cromatografía de Afinidad/métodos , Electroforesis en Gel de Poliacrilamida , Humanos , Masculino , Neoplasias Hormono-Dependientes/metabolismo , Espectrometría de Masas en Tándem
18.
Analyst ; 139(17): 4350-5, 2014 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-25010982

RESUMEN

The flowing atmospheric pressure afterglow (FAPA) ion source operates in the ambient atmosphere and has been proven to be a promising tool for direct and rapid determination of numerous compounds. Here we linked a FAPA-MS system to an electrochemical flow cell for the identification of drug metabolites generated electrochemically in order to study simulated metabolic pathways. Psychostimulants and their metabolites produced by electrochemistry (EC) were detected on-line by FAPA-MS. The FAPA source has never been used before for an on-line connection with liquid flow, neither for identification of products generated in an electrochemical flow cell. The system was optimized to achieve the highest ionization efficiency by adjusting several parameters, including distances and angles between the ion source and the outlet of the EC system, the high voltage for plasma generation, flow-rates, and EC parameters. Simulated metabolites from tested compounds [methamphetamine (MAF), para-methoxy-N-methylamphetamine (PMMA), dextromethorphan (DXM), and benzydamine (BAM)] were formed in the EC cell at various pH levels. In all cases the main products were oxidized substrates and compounds after N-demethylation. Generation of such products and their thorough on-line identification confirm that the cytochrome P450 - driven metabolism of pharmaceuticals can be efficiently simulated in an electrochemical cell; this approach may serve as a step towards predictive pharmacology using a fast and robust design.


Asunto(s)
Antiinflamatorios/análisis , Bencidamina/análisis , Estimulantes del Sistema Nervioso Central/análisis , Dextrometorfano/análisis , Antagonistas de Aminoácidos Excitadores/análisis , Espectrometría de Masas/instrumentación , Metanfetamina/análisis , Antiinflamatorios/metabolismo , Presión Atmosférica , Bencidamina/metabolismo , Estimulantes del Sistema Nervioso Central/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Dextrometorfano/metabolismo , Técnicas Electroquímicas/instrumentación , Diseño de Equipo , Antagonistas de Aminoácidos Excitadores/metabolismo , Humanos , Redes y Vías Metabólicas , Metanfetamina/análogos & derivados , Metanfetamina/metabolismo , Metilación , Oxidación-Reducción
19.
Eur J Mass Spectrom (Chichester) ; 20(4): 279-85, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25420340

RESUMEN

Knowledge of the metabolic pathways and biotransformation of the most popular drugs, such as cocaine, amphetamine, morphine and others, is crucial for the elucidation of their possible toxicity and mechanism of action in the human body. In vitro studies on metabolism are mainly based on the incubation of drugs with liver celL homogenate and utilizing Living animals. These methods need to be followed by isolation and detection of metabolic products, which makes these techniques time-consuming and technically demanding. We show here that the oxidative metabolism that occurs in the liver cells and is mainly caused by cytochrome P450 can be successfully mimicked with the electrochemical system [EC] connected on-line with electrospray ionization mass spectrometry. Cocaine was chosen as a model drug for these studies and was analyzed with a previously described system under various conditions using the boron-doped diamond working electrode. The results were compared with the number of metabolites generated by a standard procedure based on the reaction with the rat Liver microsomes. Two electrochemical products of cocaine oxidation were created, of which one was a natural metabolite of cocaine in the human body-norcocaine. The EC provides a promising platform for the screening of the addictive drug phase I metabolism. The metabolites can be directly analyzed by mass spectrometry or collected and separated by Liquid chromatog- raphy. No Liver cell homogenate or microsome is necessary to generate these metabolites, which simplifies separation of the mixtures and reduces time and costs of all experiments.


Asunto(s)
Cocaína/metabolismo , Técnicas Electroquímicas/métodos , Espectrometría de Masa por Ionización de Electrospray/métodos , Animales , Cocaína/análogos & derivados , Cocaína/análisis , Cocaína/farmacocinética , Drogas Ilícitas/farmacocinética , Masculino , Microsomas Hepáticos/metabolismo , Ratas Wistar
20.
Int J Mol Sci ; 15(9): 16787-99, 2014 Sep 22.
Artículo en Inglés | MEDLINE | ID: mdl-25247577

RESUMEN

The term "cryptome" refers to the subset of cryptic peptides with bioactivities that are often unpredictable and very different from the parent protein. These cryptic peptides are generated by proteolytic cleavage of proteases, whose identification in vivo can be very challenging. In this work, we show that insulin-degrading enzyme (IDE) is able to degrade specific amino acid sequences present in the neuropeptide pro-NPFFA (NPFF precursor), generating some cryptic peptides that are also observed after incubation with rat brain cortex homogenate. The reported experimental findings support the increasingly accredited hypothesis, according to which, due to its wide substrate selectivity, IDE is involved in a wide variety of physiopathological processes.


Asunto(s)
Corteza Cerebral/enzimología , Insulisina/fisiología , Proteínas del Tejido Nervioso/fisiología , Precursores de Proteínas/metabolismo , Receptores de Neuropéptido/metabolismo , Secuencia de Aminoácidos , Animales , Masculino , Espectrometría de Masas , Modelos Biológicos , Datos de Secuencia Molecular , Fragmentos de Péptidos/metabolismo , Proteolisis , Ratas , Ratas Wistar , Espectrometría de Masa por Ionización de Electrospray , Resonancia por Plasmón de Superficie , Extractos de Tejidos
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA