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This article describes the history, development, and current state of two analytical methods to which Czech scientists have made significant contributions. It describes the first steps of these methods in Czechoslovakia during the 1970s and 1980s, the development of the methods through innovation, and the establishment of cooperation with foreign scientists. Today, Czech scientists from the Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, and Institute of Analytical Chemistry of the Czech Academy of Sciences in Brno are among the major contributors to these fields of analytical chemistry. The milestones in the development and the prominent scientists mentioned in the individual chapters give a chance for a bright future for these methods, not only in the Czech Republic.
- MeSH
- dějiny 20. století MeSH
- dějiny 21. století MeSH
- elektroforéza kapilární * dějiny metody MeSH
- lidé MeSH
- průtoková injekční analýza MeSH
- Check Tag
- dějiny 20. století MeSH
- dějiny 21. století MeSH
- lidé MeSH
- Publikační typ
- historické články MeSH
- Geografické názvy
- Česká republika MeSH
Intoxikace metanolem může způsobit diagnostické potíže, pokud neexistuje údaj o požití závadné látky. V úvodu epidemie metanolových otrav v ČR v roce 2012 jsme se s negativní anamnézou konzumace alkoholu potýkali hned několikrát. Hladiny metanolu byly u těchto několika pacientů stopové, někdy dokonce nulové. Pacienti trpěli četnými přidruženými nemocemi, někteří z nich museli být během přijetí resuscitováni. Jednoduchá metoda stanovení hladiny kyseliny mravenčí nebyla v prvních dnech vlny otrav běžně dostupná. Intoxikace metanolem byla definitivně potvrzena v těchto případech až dodatečným vyšetřením kyseliny mravenčí v séru ve specializované laboratoři s odstupem několika dalších dnů. Tyto skutečnosti nás vedly k ověření možnosti detekce nadlimitní koncentrace formiátu v séru rychlým screeningovým vyšetřením. Analýza vzorků krevního séra, odebraných během léčby intoxikace metanolem, kapilární elektroforézou ukázala, že lze dobře monitorovat hladinu formiátu v séru jak před zahájením terapie, tak během ní. Klíčová slova: metanol – kyselina mravenčí – acidóza – kapilární elektroforéza – hemodialýza
Methanol intoxication can be difficult to diagnose, especially if there is no supporting evidence of ingestion of the toxic compound. At the begining of the methanol intoxication epidemic in the Czech Republic (2012) we faced a negative anamnesis of alcohol consumption several times. The measured levels of methanol in these patients were very low, sometimes methanol was even not detected. The patients were suffering from concomitant illnesses and some of them had to be resuscitated during admission. Simple screening methods able to detect formic acid were not available at the beginning of the epidemic. Eventually, methanol intoxication was confirmed a few days later in a specialized toxicological laboratory. Therefore, we tested a novel capillary electrophoretic method for the rapid screening of concentration of formic acid in the serum above its normal physiological range. Analysis of the serum of the patients intoxicated with methanol proved that monitoring of formate concentration in the serum with the newly developed method before and during the medical management is possible. Keywords: methanol – formic acid – acidosis – capillary electrophoresis – haemodialysis
- Klíčová slova
- kyselina mravenčí,
- MeSH
- acidóza etiologie patofyziologie terapie MeSH
- antidota terapeutické užití MeSH
- časná diagnóza MeSH
- časové faktory MeSH
- dialýza ledvin MeSH
- elektroforéza kapilární * metody statistika a číselné údaje využití MeSH
- ethanol aplikace a dávkování terapeutické užití MeSH
- fomepizol MeSH
- formiáty * krev metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- methanol * krev metabolismus otrava MeSH
- otrava diagnóza patofyziologie terapie MeSH
- péče o pacienty v kritickém stavu MeSH
- pyrazoly terapeutické užití MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
Scaffolds modified with nanostructures are recently finding use in a broad range of applications spanning from chromatographic separations to tissue engineering. This continuation of the review series on design and applications of monolithic materials covers some of the less common monoliths including use of nanostructures in preparation, modifications, and applications.
- MeSH
- chromatografie přístrojové vybavení metody MeSH
- DNA izolace a purifikace MeSH
- lidé MeSH
- nanostruktury chemie MeSH
- peptidy izolace a purifikace MeSH
- proteiny izolace a purifikace MeSH
- syntetické pryskyřice chemie MeSH
- tkáňové inženýrství přístrojové vybavení MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
We report an efficient and streamlined way to improve the analysis and identification of peptides and proteins in complex mixtures of soluble proteins, cell lysates, etc. By using the shotgun proteomics methodology combined with bioaffinity purification we can remove or minimize the interference contamination of a complex tryptic digest and so avoid the time-consuming separation steps before the final MS analysis. We have proved that by means of enzymatic fragmentation (endoproteinases with Arg-C or/and Lys-C specificity) connected with the isolation of specific peptides we can obtain a simplified peptide mixture for easier identification of the entire protein. A new bioaffinity sorbent was developed for this purpose. Anhydrotrypsin (AHT), an inactive form of trypsin with an affinity for peptides with arginine (Arg) or lysine (Lys) at the C-terminus, was immobilized onto micro/nanoparticles with superparamagnetic properties (silica magnetite particles (SiMAG)-Carboxyl, Chemicell, Germany). This AHT carrier with a determined binding capacity (26.8 nmol/mg of carrier) was tested with a model peptide, human neurotensin, and the resulting MS spectra confirmed the validity of this approach.
- MeSH
- bioreaktory MeSH
- časové faktory MeSH
- chromatografie afinitní metody přístrojové vybavení MeSH
- enzymy imobilizované chemie MeSH
- financování organizované MeSH
- lidé MeSH
- ligandy MeSH
- magnetismus MeSH
- metaloendopeptidasy chemie MeSH
- nanočástice chemie MeSH
- neurotensin analýza MeSH
- oxid křemičitý MeSH
- peptidy analýza chemie MeSH
- proteomika MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- serinové endopeptidasy chemie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- trypsin chemie izolace a purifikace MeSH
- Check Tag
- lidé MeSH
Open tubular capillary enzyme reactors were studied for rapid protein digestion and possible on-line integration into a CE/ESI/MS system. The need to minimize the time of the analyte molecules to diffuse towards the surface immobilized enzyme and to maximize the surface-to-volume (S/V) ratio of the open tubular reactors dictated the use of very narrow bore capillaries. Extremely small protein amounts (atto-femtomoles loaded) could be digested with enzymes immobilized directly on the inside wall of a 10 microm I.D. capillary. Covalently immobilized L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK)-trypsin and pepsin A were tested for the surface immobilization. The enzymatic activity was characterized in the flow-through mode with on-line coupling to electrospray ionization-time of flight-mass spectrometer (ESI/TOF-MS) under a range of protein concentrations, buffer pH's, temperatures and reaction times. The optimized reactors were tested as the nanospray needles for fast identification of proteins using CE-ESI/TOF-MS.
- MeSH
- biokompatibilní potahované materiály MeSH
- elektroforéza kapilární metody MeSH
- elektroforéza mikročipová MeSH
- elektrolyty MeSH
- enzymy imobilizované chemická syntéza klasifikace MeSH
- financování organizované MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- mikrochemie MeSH
- on-line systémy MeSH
- pepsin A metabolismus MeSH
- proteiny analýza chemie MeSH
- sekvenční analýza proteinů metody MeSH
- senzitivita a specificita MeSH
- studie proveditelnosti MeSH
- trypsin metabolismus MeSH
A pressurized liquid junction nanoelectrospray interface was designed and optimized for reliable on-line CE-MS coupling. The system was constructed as an integrated device for highly sensitive and selective analyses of proteins and peptides with the separation and spray capillaries fixed in a pressurized spray liquid reservoir equipped with the electrode for connection of the electrospray potential. The electrode chamber on the injection side of the separation capillary and the spray liquid reservoir were pneumatically connected by a Teflon tube filled with pressurized nitrogen. This arrangement provided precisely counterbalanced pressures at the inlet and outlet of the separation capillary. The pressure control system was driven by an electrically operated valve and maintained the optimum flow rate for the electrospray stability. All parts of the interface being in contact with the CEBGE, spray liquid and/or sample were made of glass or Teflon. The use of these materials minimized the electrospray chemical noise often caused by plastic softeners or material degradation. During optimization, the transfer of the separated zones between the separation and electrospray capillaries was monitored by UV absorbance and contactless conductivity detectors placed at the outlet of the separation capillary and inlet of the electrospray tip, respectively. This arrangement allowed independent monitoring of the effects of pressure, CE voltage and geometry of the liquid junction on the spreading and dilution of the separated zones after passage through the interface.
