Specific salts effect is intensively studied from the prospective of modification of different physico-chemical properties of biomacromolecules. Limited knowledge of the specific salts effect on enzymes led us to address the influence of five sodium anions: sulfate, phosphate, chloride, bromide, and perchlorate, on catalytic and conformational properties of human rhinovirus-14 (HRV) 3C protease. The enzyme conformation was monitored by circular dichroism spectrum (CD) and by tyrosines fluorescence. Stability and flexibility of the enzyme have been analyzed by CD in the far-UV region, differential scanning calorimetry and molecular dynamics simulations, respectively. We showed significant influence of the anions on the enzyme properties in accordance with the Hofmeister effect. The HRV 3C protease in the presence of kosmotropic anions, in contrast with chaotropic anions, exhibits increased stability, rigidity. Correlations of stabilization effect of anions on the enzyme with their charge density and the rate constant of the enzyme with the viscosity B-coefficients of anions suggest direct interaction of the anions with HRV 3C protease. The role of stabilization and decreased fluctuation of the polypeptide chain of HRV 3C protease on its activation in the presence of kosmotropic anions is discussed within the frame of the macromolecular rate theory.
- MeSH
- anionty chemie MeSH
- lidé MeSH
- prospektivní studie MeSH
- proteasy * MeSH
- soli * chemie MeSH
- srdeční frekvence MeSH
- virové proteasy 3C MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A set of new mixed-mode ion-exchange stationary phases is presented. The backbone of organic selectors is formed by a linear hydrocarbon chain, which is divided into two parts of various lengths by a heteroatom (oxygen or nitrogen). In all studied cases, there is a sulfonic acid moiety as the terminal group. Therefore, selectors bearing oxygen gave rise to strong cation ion-exchange stationary phases, while selectors with an embedded nitrogen atom (inducing a weak anion exchange capacity) were used to create zwitterion ion-exchange stationary phases. The new mixed-mode stationary phases were chromatographically evaluated in high performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) using isocratic elution conditions to disclose their chromatographic potential. In HPLC mode, aqueous-rich reversed phase chromatography, acetonitrile-rich hydrophilic interaction liquid chromatography and methanolic ion-exchange chromatography mobile phases were employed. In these chromatographic modes, retention factors and selectivity values for a test set of basic and zwitterionic analytes were determined. The results were compared and principal component analysis for each chromatographic mode was performed. For all chromatographic modes, the component 1 in the principal component analysis reflected the elution order. The application of different mobile phases on a particular column resulted not only in variation in retention, but also in modified selectivity, and different elution order of the analytes. The orthogonality of the elution order depending on the employed mobile phase conditions was especially reflected for structurally closely related analytes, such as melatonin and N-acetyl-serotonin, tryptamine and serotonin or noradrenalin and octopamine. However, ion-exchange interactions remain the main driving force for retention. From all investigated stationary phases, the SCX 2 (C5-linker and C4-spacer) seems to be the best choice for the separation of basic analytes using different mobile phase conditions.
- MeSH
- acetonitrily chemie MeSH
- anionty chemie MeSH
- biogenní aminy izolace a purifikace MeSH
- chemie farmaceutická metody MeSH
- chromatografie metody MeSH
- hydrofobní a hydrofilní interakce MeSH
- iontová výměna MeSH
- kationty chemie MeSH
- léčivé přípravky chemie izolace a purifikace MeSH
- methanol chemie MeSH
- voda chemie MeSH
- Publikační typ
- časopisecké články MeSH
Asymmetric and symmetric dimethylarginines are toxic non-coded amino acids. They are formed by post-translational modifications and play multifunctional roles in some human diseases. Their determination in human blood plasma is performed using capillary electrophoresis with contactless conductivity detection. The separations are performed in a capillary covered with covalently bonded PAMAPTAC polymer, which generates anionic electroosmotic flow and the separation takes place in the counter-current regime. The background electrolyte is a 750 mM aqueous solution of acetic acid with pH 2.45. The plasma samples for analysis are treated by the addition of acetonitrile and injected into the capillary in a large volume, reaching 94.5% of the total volume of the capillary, and subsequently subjected to electrophoretic stacking. The attained LODs are 16 nm for ADMA and 22 nM for SDMA. The electrophoretic resolution of both isomers has a value of 5.3. The developed method is sufficiently sensitive for the determination of plasmatic levels of ADMA and SDMA. The determination does not require derivatization and the individual steps in the electrophoretic stacking are fully automated. The determined plasmatic levels for healthy individuals vary in the range 0.36-0.62 µM for ADMA and 0.32-0.70 µM for SDMA.
In this work, two commercialized anion-exchange membranes (AEMs), AMI-7001 and AF49R27, were applied in microbial electrolysis cells (MECs) and compared with a novel AEM (PSEBS CM DBC, functionalized with 1,4-diazabicyclo[2.2.2]octane) to produce biohydrogen. The evaluation regarding the effect of using different AEMs was carried out using simple (acetate) and complex (mixture of acetate, butyrate and propionate to mimic dark fermentation effluent) substrates. The MECs equipped with various AEMs were assessed based on their electrochemical efficiencies, H2 generation capacities and the composition of anodic biofilm communities. pH imbalances, ionic losses and cathodic overpotentials were taken into consideration together with changes to substantial AEM properties (particularly ion-exchange capacity, ionic conductivity, area- and specific resistances) before and after AEMs were applied in the process to describe their potential impact on the behavior of MECs. It was concluded that the MECs which employed the PSEBS CM DBC membrane provided the highest H2 yield and lowest internal losses compared to the two other separators. Therefore, it has the potential to improve MECs.
- MeSH
- anionty chemie MeSH
- design vybavení MeSH
- elektrolýza MeSH
- Geobacter metabolismus MeSH
- kvartérní amoniové sloučeniny chemie MeSH
- membrány umělé * MeSH
- piperaziny chemie MeSH
- studie proveditelnosti MeSH
- vodík metabolismus MeSH
- zdroje bioelektrické energie * mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
In this work, we have generalized the use of capillary isotachophoresis as a universal method for determination of effective charge of anionic and cationic (co)polymers on ordinary capillary electrophoresis instruments. This method is applicable to a broad range of strong or weak polyelectrolytes with good repeatability. Experimental parameters (components and concentrations of leading and terminating electrolytes, capillary diameters, constant electric current intensity) were optimized for implementation in 100 μm i.d. capillaries for both polyanions and polycations. Determined values of polymer effective charge were in a very good agreement with those obtained by capillary electrophoresis with indirect UV detection. Uncertainty of the effective charge measurement using isotachophoresis was addressed and estimated to be ∼5-10% for solutes with mobilities in the 20-50 × 10(-9)m(2)V(-1)s(-1) range.
- MeSH
- anionty chemie MeSH
- elektroforéza kapilární přístrojové vybavení metody MeSH
- elektrolyty MeSH
- izotachoforéza přístrojové vybavení metody MeSH
- kationty chemie MeSH
- koncentrace vodíkových iontů MeSH
- polymery chemie MeSH
- reprodukovatelnost výsledků MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A micro-electromembrane extraction (μ-EME) technique using electrically induced transfer of charged analytes across free liquid membranes (FLMs) was presented. A disposable extraction unit was proposed and it was made of a short segment of transparent perfluoroalkoxy tubing, which was successively filled with three liquid plugs serving as acceptor solution, FLM and donor solution. These plugs formed a three-phase extraction system, which was precisely defined, that was stable and required μL to sub-μL volumes of all respective solutions. Basic instrumental set-up and extraction principles of μ-EME were examined using an anionic and a cationic dye, 4,5-dihydroxy-3-(p-sulfophenylazo)-2,7-naphthalene disulfonic acid trisodium salt (SPADNS) and crystal violet, respectively. Transfers of the charged dyes from donor into acceptor solutions across FLMs consisting of 1-pentanol were visualized by a microscope camera and quantitative measurements were performed by UV-vis spectrophotometry. The effects of operational parameters of μ-EME system were comprehensively investigated and experimental measurements were accompanied with theoretical calculations. Extraction recoveries above 60% were achieved for 5min μ-EME of 1mM SPADNS at 100V with repeatability values below 5%. Selectivity of FLMs was additionally examined by capillary electrophoretic analyses of acceptor solutions and the potential of FLMs for μ-EME pretreatment of samples with artificial complex matrices was demonstrated.
3,5-Dinitrobenzoate and picrate are light absorbing anions pertinent to indirect photometric detection of boron cluster anions in buffered methanolic background electrolytes (BGEs). Tris(hydroxymethyl)aminomethane and morpholine have been used as buffering bases, which eliminated baseline steps, and minimized the baseline noise. In methanolic BGEs, mobilities of boron cluster anions depend on both ionic constituents of the BGE buffer. This dependence can be explained by ion pair interaction of detected anions with BGE cations, which are not bonded into ion pairs with the BGE anions. The former ion pair interaction decreases sensitivity of the indirect photometric detection.
Anion exchange polystyrene nanofiber materials (AE) were prepared by electrospinning followed by two-step functionalization of the nanofiber surface by chlorosulfonic acid and ethylendiamine. The photoactive character of these materials was introduced through adsorption of the tetra-anionic 5,10,15,20-tetrakis-(4-sulfonatophenyl)porphyrin photosensitizer (TPPS-AE) on the nanofiber surface or by encapsulation of the nonpolar 5,10,15,20-tetraphenylporphyrin photosensitizer (AE(TPP)) into the nanofibers. Anion exchange nanofiber materials with porphyrins are characterized by a high ion-exchange capacity, photogeneration of singlet oxygen O2((1)Δg), and singlet oxygen-sensitized delayed fluorescence. Due to the photogeneration of cytotoxic O2((1)Δg), the nanofibers exhibited oxidation of the external substrates in aqueous solution and an efficient antibacterial effect when activated by simulated daylight. Adsorption of both TPPS and I(-) on the surface of AE led to the formation of more efficient I-TPPS-AE materials. Rapid photooxidation of I(-) by O2((1)Δg), and the formation of another cytotoxic species, I3(-), on the surface of the nanofibers were responsible for the increased antibacterial properties of I-TPPS-AE and the prolonged antibacterial effect in the dark.
- MeSH
- adsorpce MeSH
- anionty chemie MeSH
- Escherichia coli účinky léků metabolismus účinky záření MeSH
- fotosenzibilizující látky chemie farmakologie MeSH
- iontová výměna MeSH
- kinetika MeSH
- nanovlákna chemie MeSH
- oxidace-redukce MeSH
- polystyreny chemie MeSH
- porfyriny chemie farmakologie MeSH
- singletový kyslík chemie metabolismus MeSH
- světlo * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The subject of this work is the definition of a simple model based on general ITP theory that allows describing and predicting the behavior of ITP systems compatible with ESI-MS detection. The model is exemplified by anionic ITP of weak acids that represent an interesting potential application field of ITP-ESI-MS. Suitable ESI-compatible electrolyte systems of very simple composition are proposed including a special free-acid ITP arrangement. The properties of these systems are discussed using illustrative diagrams of their stacking windows. The use of anionic ITP-ESI-MS in negative-ion ESI mode is reported for the first time and its suitability for sensitive trace analysis is demonstrated. The presented ITP-ESI-MS application example comprises a free-acid ITP system formed of formic and propionic acids and direct injection analysis of ibuprofen and diclofenac in waters with quantitation limits of the order 10(-10) M.
- MeSH
- anionty chemie MeSH
- antiflogistika nesteroidní analýza MeSH
- diklofenak analýza MeSH
- elektroforéza kapilární MeSH
- elektrolyty chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- ibuprofen analýza MeSH
- izotachoforéza metody MeSH
- kyseliny chemie MeSH
- limita detekce MeSH
- neopioidní analgetika analýza MeSH
- pitná voda analýza MeSH
- řeky chemie MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
A 2-hydroxyphenacyl moiety absorbing below 370 nm is proposed as a new photoremovable protecting group for carboxylates and sulfonates. Laser flash photolysis and steady-state sensitization studies show that the leaving group is released from a short-lived triplet state. In addition, DFT-based quantum chemical calculations were performed to determine the key reaction steps. We found that triplet excited state intramolecular proton transfer represents a major deactivation channel. Minor productive pathways involving the triplet anion and quinoid triplet enol intermediates have also been identified.
