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Development of divergent-flow isoelectric focusing chip for separation of biological agents
Duša, Filip ; Šalplachta, Jiří ; Horká, Marie ; Lunerová, K. ; Rosenbergová, K. ; Kubíček, O.
Isoelectric focusing (IEF) is an efficient method for separation of different bacterium species. Divergent flow format of IEF (DF-IEF) enables continual separation of large volumes of samples and fractionation of the separated analytes for later analysis. The mentioned properties enabled development of DF-IEF chip for separation of H2O2 inactivated biological agents (B-agents). The chip was designed to perform separation in vertical alignment which solved issues connected with sedimentation of bacterial cells. The separation method was optimized using colored markers of isoelectric point and four different b-agents were separated using the DF-IEF chip. The obtained fractions were subsequently analyzed with matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). The mass spectra proved that the developed DF-IEF chip is an effective tool for separation of inactivated B-agents.
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Detection and identification of aflatoxins by means of portable liquid chromatograph for purpose of control of biological weapons prohibition
Šesták, Jozef ; Lunerová, K.
Method describes screening detection and identification of selected mycotoxins aflatoxins (aflatoxins B1, B2, G1 and G2), ochratoxins A, B and zearalenon by means of portable miniaturzed capillary liquid chromatograph (miniLC) with UV/VIS and fluorescence detector (FLD). The aim of the method is to provide rapid screening method for finding/excluding the presence of given mycotoxins in unknown samples. In case of positive finding, it is necessary to perform confirmation analysis using other instrumental methods (e. g. LC-MS). The method is intended namely for the needs of technical support of the State Office for Nuclear Safety for control and supervising activities in field of execution of the Act No. 281/1997 Coll (Biological Act).
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Evaluation of the 33 synthesized nitrophenol-based markers of isoelectic point
Duša, Filip ; Moravcová, Dana ; Šlais, Karel
33 nitrophenol-based compounds, which absorb at ultraviolet as well as visible wavelengths, were selected as candidates for marking isoelectric points (pI) over the pH range from 3 to 10. The evaluation and selection of the candidate structures included structure-based calculations of acid-base properties. The selected compounds were further tested experimentally using capillary isoelectric focusing. The experimental analysis excluded two compounds due to producing double peaks during the analysis. Overall, 31 novel compounds were identified as suitable pI markers.
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