UV–vis detection Dotaz Zobrazit nápovědu
Hemoglobinopathies, hereditary disorders affecting the structure or production of hemoglobin, were detected by routine HbA1c measurements by capillary electrophoresis (CE) at the University Hospital Motol, Prague. The potential of ultraviolet-visible (UV-Vis) and Fourier-transform infrared (FTIR) spectroscopy for the detection and characterization of hemoglobinopathies was investigated. FTIR spectra were recorded with a very high resolution (0.5 cm-1) with 128 scans. The broad amide I peak, located at 1700-1600 cm-1, can be formed by superimposition of the conformational structures of hemoglobin. These secondary protein structures were subjected to mathematical analysis. The application of band narrowing techniques, followed by curve fitting and integration processes, provided the basis for the quantitative estimation of protein secondary structure. As a result, unambiguous differences in UV-Vis spectra among patients with presumably normal hemoglobin, an HbC or a hemoglobin S/hemoglobin G (HbS/HbG)-Philadelphia variant could not be demonstrated. However, FTIR spectra indicated slight differences in α-helix, β-turns, β-sheet, or random coil secondary hemoglobin structures for these mutations. In the spectral wavenumber range of 950-850 cm-1, there were some obvious FTIR differences at specific wavenumbers between patients with normal hemoglobin and those with the HbC variant. Further investigations are needed with a sufficient number of hemoglobin variants to elucidate the potency of FTIR spectroscopy for the characterization of hemoglobinopathies.
- Klíčová slova
- Fourier‐transform infrared (FTIR) spectroscopy, capillary electrophoresis, hemoglobinopathy, ultraviolet–visible (UV–Vis),
- Publikační typ
- časopisecké články MeSH
In this work, we describe the introduction of a post-column solid-state reactor in the HPLC system used for the analyses of amino acids. The reactor used was filled with copper(II) oxide. Passage of the analytes through the reactor leads to the formation of Cu(II) complexes. Unlike free amino acids, the Cu-complexes show significant absorbance in the UV region and accordingly sensitivity of UV-VIS detection is increased by two to three orders of magnitude. As a result of this improvement in sensitivity, we have obtained LOD values in micromolar range and good linearity over the studied concentration range (5.0×10(-5) to 2.0×10(-3) mol/L). The method exhibits advantages typical of solid-state reactors, such as negligible loss of efficiency due to the derivatization, simplicity of realization and a long-term durability. The presented system brings an easy and versatile solution for UV-VIS detection of coordinating compounds, which do not normally absorb well in the UV-VIS region.
- MeSH
- aminokyseliny chemie MeSH
- měď chemie MeSH
- spektrofotometrie ultrafialová MeSH
- spektrofotometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aminokyseliny MeSH
- cupric oxide MeSH Prohlížeč
- měď MeSH
Constraining the formation mechanisms of organic matter that persists in aquatic reservoirs is important for determining the reactivity and fate of carbon and nutrients in these environments. Recent studies have linked dissolved organic matter (DOM) accumulating in the ocean to linear terpenoid structures, and carotenoid degradation products have been proposed as potential precursors. The prevalence of reactive oxygen species in aquatic environments and their potential to be quenched by carotenoids led us to examine radical-assisted photochemical degradation of carotenoids as a potential mechanism for DOM formation and transformation. Experiments were conducted with aggregates of β-carotene, astaxanthin, fucoxanthin and meso-zeaxanthin in THF:H2O under solar light irradiation assisted by hydrogen peroxide (UV-Vis/H2O2). Based on the fine structure of UV-Vis spectra, it was determined that β-carotene and meso-zeaxanthin formed J-type aggregates in experimental solutions, while astaxanthin and fucoxanthin formed H2-type aggregates, consistent with their structural characteristics. All carotenoids degraded under the combined influence of photolysis and OH scavenging, with fucoxanthin exhibiting the fastest degradation kinetics (kPO = 3.69 10-3 s-1) and meso-zeaxanthin the slowest (kPO = 4.37 10-4 s-1). The major degradation products detected by electrospray ionization (ESI) tandem mass spectrometry (MS/MS) were apo-aldehydes and apo-ketones, with the latter tending to accumulate, but epoxidation of the carotenoids also took place, and longer irradiation times resulted in lower molecular weight products. Reaction kinetics and accumulating carotenoid oxidation products identified in this study provide potential formation mechanisms and biomarkers for examining DOM cycling.
- Klíčová slova
- Aggregates, Carotenoids, Kinetics, Oxidation products, Solar irradiation,
- MeSH
- beta-karoten MeSH
- karotenoidy * MeSH
- kinetika MeSH
- peroxid vodíku * MeSH
- tandemová hmotnostní spektrometrie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- beta-karoten MeSH
- karotenoidy * MeSH
- peroxid vodíku * MeSH
A novel colorimetric sensor based on the TiO2/poly(acrylamide-co-methylene bis acrylamide-co-2-(3-(4-nitro-phenyl)thioureido)ethyl methacrylate) nanocomposite was synthesized via a surface modification strategy; methacryloxypropyltrimethoxysilane was used to provide reactive vinyl groups on the surface of TiO2 nanoparticles for the successful surface polymerization of Am (acrylamide), MBA (methylenbisacrylamide), and NPhM (2-(3-(4-nitrophenyl)thioureido)ethyl methacrylate) components. The successful preparation of nanocomposites was confirmed using Fourier transform infrared, 1H NMR, 13C NMR, scanning electron microscopy, transmission electron microscopy, thermogravimetry analysis, and X-ray diffraction methods, and the sensing ability of the probe toward fluoride ions was investigated using naked-eye detection and UV-vis measurement. The interaction of the prepared polymeric nanocomposite with fluoride ions elicited a significant visible change in color from pale yellow to orange and was further affirmed by a clean interconversion of the two absorption bands at 330 and 485 nm. The selective binding ability of the polymeric nanocomposite towards fluoride over other anions, such as I-, Cl-, Br-, AcO-, H2PO4 -, and H2SO4 - was further explored; the prepared chemosensor could detect fluoride ions in acetonitrile with a detection limit of 3 μM.
- Publikační typ
- časopisecké články MeSH
Using solid-state 15N NMR spectroscopy, the cis/trans isomerization in a two-dimensional (2-D) array of surface-mounted azobenzene-based switches was detected for the first time. In order to achieve this, a new class of rod-shaped molecular switches, suitable for formation of 2-D regular arrays on large facets of tris(o-phenylenedioxy)cyclotriphosphazene (TPP) nanocrystals, was synthesized. A mechanochemical approach was used to prepare corresponding host-guest surface inclusions in a TPP matrix. Comparison of thermal steps in solution and supramolecular surface inclusions revealed that switching of individual molecules is not compromised by the close proximity of neighbors.
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The analysis of dietary supplements is far less regulated than pharmaceuticals, leading to potential quality issues. Considering their positive effect, many athletes consume supplements containing L-histidine and β-alanine. A new microfluidic method for the determination of L-histidine and β-alanine in dietary supplement formulations has been developed. For the first time, capacitively coupled contactless conductivity detection was employed for the microchip electrophoresis of amino acids in real samples. A linear relationship between detector response and concentration was observed in the range of 10-100 µmol L-1 for L-histidine (R2 = 0.9968) and β-alanine (R2 = 0.9954), while achieved limits of detection (3 × S/N ratio) were 4.2 µmol L-1 and 5.2 µmol L-1, respectively. The accuracy of the method was confirmed using recovery experiments as well as CE-UV-VIS and HPLC-UV-VIS techniques. The developed method allows unambiguous identification of amino acids in native form without chemical derivatization and with the possibility of simultaneous analysis of amino acids with metal cations.
- Klíčová slova
- l-Histidine, Amino acids, Capacitively coupled contactless conductivity detection, Dietary supplements, Microchip electrophoresis, β-Alanine,
- MeSH
- beta-alanin * analýza chemie MeSH
- elektrická vodivost * MeSH
- elektroforéza mikročipová * metody MeSH
- histidin * analýza chemie MeSH
- limita detekce MeSH
- potravní doplňky * analýza MeSH
- sklo chemie MeSH
- technologie zelené chemie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- beta-alanin * MeSH
- histidin * MeSH
A simple sample injection procedure compatible with commercial capillary electrophoresis (CE) instrumentation was developed, which enables handling sample volumes as little as 250nL for analytical applications where sample volume availability is of concern. Single-use micro-sampling inserts were prepared by thermal modification of polypropylene micropipette tips and the inserts were accommodated in standard CE vials in CE autosampler carousel. To ensure direct contact of separation capillary injection end with sample solution and to avoid possible damage to the capillary, a soft compression spring was placed at the bottom of the vial underneath the micro-sampling insert. Injections from sub-μL samples were carried out in conventional as well as in short-end injection mode, were compatible with standard i.d./o.d. (25-100μm/365μm) fused silica capillaries and with various background electrolyte solutions and detection modes. Excellent repeatability of replicate injections from 250nL to 3μL was achieved based on RSD values of quantitative analytical measures (peak heights ≤2.4% and peak areas ≤3.7%) for CE-UV-vis, CE-ESI-MS and CE-contactless conductivity detection of model basic drugs. The achieved RSD values were comparable with those for replicate injections of the drugs from standard CE vials. The reported concept of injections from micro-sampling inserts was further demonstrated useful in evaluation of micro-electromembrane extraction (μ-EME) of model basic drugs. Sub-μL volumes of operational solutions resulted in reduced lengths of μ-EME phases and improved extraction recoveries (66-91%) were achieved.
- Klíčová slova
- Capillary electrophoresis, Contactless conductivity detection, Mass spectrometry, Micro-extractions, Micro-sampling, UV–vis detection,
- MeSH
- elektroforéza kapilární přístrojové vybavení metody MeSH
- elektrolyty chemie MeSH
- hmotnostní spektrometrie MeSH
- nortriptylin analýza izolace a purifikace MeSH
- papaverin analýza MeSH
- roztoky chemie MeSH
- spektrofotometrie ultrafialová MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- elektrolyty MeSH
- nortriptylin MeSH
- papaverin MeSH
- roztoky MeSH
Preconcentration potential of micro-electromembrane extraction (μ-EME) across free liquid membrane (FLM) was examined with an anionic and a cationic dye, 4,5-dihydroxy-3-(p-sulfophenylazo)-2,7-naphthalene disulfonic acid, trisodium salt (SPADNS) and phenosafranine, respectively. For the first time, it was shown that the spatial flexibility of FLMs enabled application of tailored extraction units with mutually different shapes and migration cross-sections for FLMs, donor and acceptor solutions. Thus, e.g. conical units enabled easy and reproducible formation of a three-phase extraction system (donor/FLM/acceptor) with sub-μL volumes of acceptor solutions as well as rapid and highly efficient preconcentration of the two dyes. Quantitative measurements of resulting solutions were carried out by UV-vis spectrophotometry and enrichment factors of up to 98 were achieved for μ-EMEs of 20 μM SPADNS (50 μL) preconcentrated into 0.5 μL of pure water across 1-pentanol at -150 V for 18 min. Visual monitoring of the entire extraction process (with USB microscope camera) was possible across transparent extraction units, moreover, important extraction parameters, such as FLM dimensions and donor-to-acceptor solution volume ratio, which determine the mechanical stability of the membrane and maximum enrichment factor, respectively, were readily adjusted. Combination of μ-EME across FLMs with capillary electrophoresis (CE) was further shown suitable for preconcentration and determination of perchlorate in drinking water samples. Good repeatability of the μ-EME-CE method (RSD values better than 9.5%), linear relationship for the analytical signal vs. concentration (r(2) better than 0.997) and enrichment factors of up to 30 were achieved for μ-EMEs of perchlorate across 1-pentanol and 1-hexanol based FLMs.
Lactoferrin is a multifunctional protein with antimicrobial activity and others tohealth beneficial properties. The main aim of this work was to propose easy to usetechnique for lactoferrin isolation from cow colostrum samples. Primarily we utilizedsodium dodecyl sulphate - polyacrylamide gel electrophoresis for isolation of lactoferrinfrom the real samples. Moreover we tested automated microfluidic Experionelectrophoresis system to isolate lactoferrin from the collostrum sample. The welldeveloped signal of lactoferrin was determined with detection limit (3 S/N) of 20 ng/ml. Inspite of the fact that Experion is faster than SDS-PAGE both separation techniques cannotbe used in routine analysis. Therefore we have tested third separation technique, ionexchange chromatography, using monolithic column coupled with UV-VIS detector (LCUV-VIS). We optimized wave length (280 nm), ionic strength of the elution solution (1.5M NaCl) and flow rate of the retention and elution solutions (0.25 ml/min and 0.75 ml/min.respectively). Under the optimal conditions the detection limit was estimated as 0.1 μg/mlof lactoferrin measured. Using LC-UV-VIS we determined that lactoferrin concentrationvaried from 0.5 g/l to 1.1 g/l in cow colostrums collected in the certain time interval up to 72 hours after birth. Further we focused on miniaturization of detection device. We testedamperometric detection at carbon electrode. The results encouraged us to attempt tominiaturise whole detection system and to test it on analysis of real samples of humanfaeces, because lactoferrin level in faeces is closely associated with the inflammations ofintestine mucous membrane. For the purpose of miniaturization we employed thetechnology of printed electrodes. The detection limit of lactoferrin was estimated as 10μg/ml measured by the screen-printed electrodes fabricated by us. The fabricatedelectrodes were compared with commercially available ones. It follows from the obtainedresults that the responses measured by commercial electrodes are app. ten times highercompared with those measured by the electrodes fabricated by us. This phenomenonrelates with smaller working electrode surface area of the electrodes fabricated by us(about 50 %) compared to the commercial ones. The screen-printed electrodes fabricatedby us were utilized for determination of lactoferrin faeces. Regarding to fact that sample offaeces was obtained from young and healthy man the amount of lactoferrin in sample wasunder the limit of detection of this method.
This work examines the effect of thermal modification temperature (180, 200, and 220 °C) in comparison with reference (untreated) samples on selected optical properties of six tropical wood species-Sp. cedar (Cedrala odorata), iroko (Chlorophora excelsa), merbau (Intsia spp.), meranti (Shorea spp.), padouk (Pterocarpus soyauxii), and teak (Tectona grandis). The main goal is to expand the existing knowledge in the field of wood thermal modification by understanding the related degradation mechanisms associated with the formation of chromophoric structures and, above all, to focus on the change in the content of extractive substances. For solid wood, the CIELAB color space parameters (L*, a*, b*, and ΔE*), yellowness (Y), ISO brightness, and UV-Vis diffuse reflectance spectra were obtained. Subsequently, these wood samples were extracted into three individual solvents (acetone, ethanol, and ethanol-toluene). The yields of the extracted compounds, their absorption spectra, and again L*, a*, b*, ΔE*, and Yi parameters were determined. With increasing temperatures, the samples lose brightness and darken, while their total color difference grows (except merbau). The highest yield of extractives (mainly phenolic compounds, glycosides, and dyes) from thermally modified samples was usually obtained using ethanol. New types of extractives (e.g., 2-furaldehyde, lactones, formic acid, some monomer derivatives of phenols, etc.) are already created around a temperature of 180 °C and may undergo condensation reactions at higher temperatures. For padouk, merbau, teak, and partially iroko modified at temperatures of 200 and 220 °C, there was a detected similarity in the intensities of their UV-Vis DR spectra at the wavelength regions corresponding to phenolic aldehydes, unsaturated ketones, quinones, stilbenes, and other conjugated carbonyl structures. Overall, a statistical assessment using PCA sorted the samples into five clusters. Cluster 3 consists of almost all samples modified at 200 and 220 °C, and in the other four, the reference and thermally modified samples at 180 °C were distributed. The yellowness of wood (Y) has a very high dependence (r = 0.972) on its brightness (L*) and the yellowness index of the extractives in acetone Yi(Ac), whose relationship was described by the equation Y = -0.0951 × Y(Ac) + 23.3485.
- Klíčová slova
- ThermoWood, UV-Vis diffuse reflectance, chemical degradation, extractives, optical properties, total color difference,
- Publikační typ
- časopisecké články MeSH
