Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 43
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Anal Bioanal Chem ; 2020 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-32918557

RESUMO

The visible-light photocatalytic E/Z isomerization of olefins can be mediated by a wide spectrum of triplet sensitizers (photocatalysts). However, the search for the most efficient photocatalysts through screenings in photo batch reactors is material and time consuming. Capillary and microchip flow reactors can accelerate this screening process. Combined with a fast analytical technique for isomer differentiation, these reactors can enable high-throughput analyses. Ion mobility (IM) spectrometry is a cost-effective technique that allows simple isomer separation and detection on the millisecond timescale. This work introduces a hyphenation method consisting of a microchip reactor and an infrared matrix-assisted laser desorption ionization (IR-MALDI) ion mobility spectrometer that has the potential for high-throughput analysis. The photocatalyzed E/Z isomerization of ethyl-3-(pyridine-3-yl)but-2-enoate (E-1) as a model substrate was chosen to demonstrate the capability of this device. Classic organic triplet sensitizers as well as Ru-, Ir-, and Cu-based complexes were tested as catalysts. The ionization efficiency of the Z-isomer is much higher at atmospheric pressure which is due to a higher proton affinity. In order to suppress proton transfer reactions by limiting the number of collisions, an IM spectrometer working at reduced pressure (max. 100 mbar) was employed. This design reduced charge transfer reactions and allowed the quantitative determination of the reaction yield in real time. Among 14 catalysts tested, four catalysts could be determined as efficient sensitizers for the E/Z isomerization of ethyl cinnamate derivative E-1. Conversion rates of up to 80% were achieved in irradiation time sequences of 10 up to 180 s. With respect to current studies found in the literature, this reduces the acquisition times from several hours to only a few minutes per scan.

2.
Anal Bioanal Chem ; 412(22): 5247-5260, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32488389

RESUMO

A new ion mobility (IM) spectrometer, enabling mobility measurements in the pressure range between 5 and 500 mbar and in the reduced field strength range E/N of 5-90 Td, was developed and characterized. Reduced mobility (K0) values were studied under low E/N (constant value) as well as high E/N (deviation from low field K0) for a series of molecular ions in nitrogen. Infrared matrix-assisted laser desorption ionization (IR-MALDI) was used in two configurations: a source working at atmospheric pressure (AP) and, for the first time, an IR-MALDI source working with a liquid (aqueous) matrix at sub-ambient/reduced pressure (RP). The influence of RP on IR-MALDI was examined and new insights into the dispersion process were gained. This enabled the optimization of the IM spectrometer for best analytical performance. While ion desolvation is less efficient at RP, the transport of ions is more efficient, leading to intensity enhancement and an increased number of oligomer ions. When deciding between AP and RP IR-MALDI, a trade-off between intensity and resolving power has to be considered. Here, the low field mobility of peptide ions was first measured and compared with reference values from ESI-IM spectrometry (at AP) as well as collision cross sections obtained from molecular dynamics simulations. The second application was the determination of the reduced mobility of various substituted ammonium ions as a function of E/N in nitrogen. The mobility is constant up to a threshold at high E/N. Beyond this threshold, mobility increases were observed. This behavior can be explained by the loss of hydrated water molecules.

3.
Sensors (Basel) ; 20(2)2020 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-31940811

RESUMO

Precision agriculture (PA) strongly relies on spatially differentiated sensor information. Handheld instruments based on laser-induced breakdown spectroscopy (LIBS) are a promising sensor technique for the in-field determination of various soil parameters. In this work, the potential of handheld LIBS for the determination of the total mass fractions of the major nutrients Ca, K, Mg, N, P and the trace nutrients Mn, Fe was evaluated. Additionally, other soil parameters, such as humus content, soil pH value and plant available P content, were determined. Since the quantification of nutrients by LIBS depends strongly on the soil matrix, various multivariate regression methods were used for calibration and prediction. These include partial least squares regression (PLSR), least absolute shrinkage and selection operator regression (Lasso), and Gaussian process regression (GPR). The best prediction results were obtained for Ca, K, Mg and Fe. The coefficients of determination obtained for other nutrients were smaller. This is due to much lower concentrations in the case of Mn, while the low number of lines and very weak intensities are the reason for the deviation of N and P. Soil parameters that are not directly related to one element, such as pH, could also be predicted. Lasso and GPR yielded slightly better results than PLSR. Additionally, several methods of data pretreatment were investigated.

4.
J Mass Spectrom ; 55(5): e4501, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31945247

RESUMO

The contamination of barley by molds on the field or in storage leads to the spoilage of grain and the production of mycotoxins, which causes major economic losses in malting facilities and breweries. Therefore, on-site detection of hidden fungus contaminations in grain storages based on the detection of volatile marker compounds is of high interest. In this work, the volatile metabolites of 10 different fungus species are identified by gas chromatography (GC) combined with two complementary mass spectrometric methods, namely, electron impact (EI) and chemical ionization at atmospheric pressure (APCI)-mass spectrometry (MS). The APCI source utilizes soft X-radiation, which enables the selective protonation of the volatile metabolites largely without side reactions. Nearly 80 volatile or semivolatile compounds from different substance classes, namely, alcohols, aldehydes, ketones, carboxylic acids, esters, substituted aromatic compounds, alkenes, terpenes, oxidized terpenes, sesquiterpenes, and oxidized sesquiterpenes, could be identified. The profiles of volatile and semivolatile metabolites of the different fungus species are characteristic of them and allow their safe differentiation. The application of the same GC parameters and APCI source allows a simple method transfer from MS to ion mobility spectrometry (IMS), which permits on-site analyses of grain stores. Characterization of IMS yields limits of detection very similar to those of APCI-MS. Accordingly, more than 90% of the volatile metabolites found by APCI-MS were also detected in IMS. In addition to different fungus genera, different species of one fungus genus could also be differentiated by GC-IMS.

5.
Sensors (Basel) ; 19(23)2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31795286

RESUMO

The lack of soil data, which are relevant, reliable, affordable, immediately available, and sufficiently detailed, is still a significant challenge in precision agriculture. A promising technology for the spatial assessment of the distribution of chemical elements within fields, without sample preparation is laser-induced breakdown spectroscopy (LIBS). Its advantages are contrasted by a strong matrix dependence of the LIBS signal which necessitates careful data evaluation. In this work, different calibration approaches for soil LIBS data are presented. The data were obtained from 139 soil samples collected on two neighboring agricultural fields in a quaternary landscape of northeast Germany with very variable soils. Reference analysis was carried out by inductively coupled plasma optical emission spectroscopy after wet digestion. The major nutrients Ca and Mg and the minor nutrient Fe were investigated. Three calibration strategies were compared. The first method was based on univariate calibration by standard addition using just one soil sample and applying the derived calibration model to the LIBS data of both fields. The second univariate model derived the calibration from the reference analytics of all samples from one field. The prediction is validated by LIBS data of the second field. The third method is a multivariate calibration approach based on partial least squares regression (PLSR). The LIBS spectra of the first field are used for training. Validation was carried out by 20-fold cross-validation using the LIBS data of the first field and independently on the second field data. The second univariate method yielded better calibration and prediction results compared to the first method, since matrix effects were better accounted for. PLSR did not strongly improve the prediction in comparison to the second univariate method.

6.
Anal Bioanal Chem ; 411(30): 8053-8061, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31741006

RESUMO

Acoustically levitated droplets have been suggested as compartmentalized, yet wall-less microreactors for high-throughput reaction optimization purposes. The absence of walls is envisioned to simplify up-scaling of the optimized reaction conditions found in the microliter volumes. A consequent pursuance of high-throughput chemistry calls for a fast, robust and sensitive analysis suited for online interrogation. For reaction optimization, targeted analysis with relatively low sensitivity suffices, while a fast, robust and automated sampling is paramount. To follow this approach, in this contribution, a direct coupling of levitated droplets to a homebuilt ion mobility spectrometer (IMS) is presented. The sampling, transfer to the gas phase, as well as the ionization are all performed by a single exposure of the sampling volume to the resonant output of a mid-IR laser. Once formed, the nascent spatially and temporally evolving analyte ion cloud needs to be guided out of the acoustically confined trap into the inlet of the ion mobility spectrometer. Since the IMS is operated at ambient pressure, no fluid dynamic along a pressure gradient can be employed. Instead, the transfer is achieved by the electrostatic potential gradient inside a dual ring electrode ion optics, guiding the analyte ion cloud into the first stage of the IMS linear drift tube accelerator. The design of the appropriate atmospheric pressure ion optics is based on the original vacuum ion optics design of Wiley and McLaren. The obtained experimental results nicely coincide with ion trajectory calculations based on a collisional model. Graphical Abstract.

7.
J Sep Sci ; 39(24): 4756-4764, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27805770

RESUMO

The combination of high-performance liquid chromatography and electrospray ionization ion mobility spectrometry facilitates the two-dimensional separation of complex mixtures in the retention and drift time plane. The ion mobility spectrometer presented here was optimized for flow rates customarily used in high-performance liquid chromatography between 100 and 1500 µL/min. The characterization of the system with respect to such parameters as the peak capacity of each time dimension and of the 2D spectrum was carried out based on a separation of a pesticide mixture containing 24 substances. While the total ion current chromatogram is coarsely resolved, exhibiting coelutions for a number of compounds, all substances can be separately detected in the 2D plane due to the orthogonality of the separations in retention and drift dimensions. Another major advantage of the ion mobility detector is the identification of substances based on their characteristic mobilities. Electrospray ionization allows the detection of substances lacking a chromophore. As an example, the separation of a mixture of 18 amino acids is presented. A software built upon the free mass spectrometry package OpenMS was developed for processing the extensive 2D data. The different processing steps are implemented as separate modules which can be arranged in a graphic workflow facilitating automated processing of data.

8.
Anal Bioanal Chem ; 408(23): 6259-68, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27370689

RESUMO

The novel combination of infrared matrix-assisted laser dispersion and ionization (IR-MALDI) with ion mobility (IM) spectrometry makes it possible to investigate biomolecules in their natural environment, liquid water. As an alternative to an ESI source, the IR-MALDI source was implemented in an in-house-developed ion mobility (IM) spectrometer. The release of ions directly from an aqueous solution is based on a phase explosion, induced by the absorption of an IR laser pulse (λ = 2.94 µm, 6 ns pulse width), which disperses the liquid as nano- and micro-droplets. The prerequisites for the application of IR-MALDI-IM spectrometry as an analytical method are narrow analyte ion signal peaks for a high spectrometer resolution. This can only be achieved by improving the desolvation of ions. One way to full desolvation is to give the cluster ions sufficient time to desolvate. Two methods for achieving this are studied: the implementation of an additional drift tube, as in ESI-IM-spectrometry, and the delayed extraction of the ions. As a result of this optimization procedure, limits of detection between 5 nM and 2.5 µM as well as linear dynamic ranges of 2-3 orders of magnitude were obtained for a number of substances. The ability of this method to analyze simple mixtures is illustrated by the separation of two different surfactant mixtures.


Assuntos
Espectrometria de Mobilidade Iônica/instrumentação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentação , Desenho de Equipamento , Raios Infravermelhos , Espectrometria de Mobilidade Iônica/métodos , Íons/química , Lasers , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
9.
J Mass Spectrom ; 51(8): 566-577, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28239970

RESUMO

A promising replacement for the radioactive sources commonly encountered in ion mobility spectrometers is a miniaturized, energy-efficient photoionization source that produce the reactant ions via soft X-radiation (2.8 keV). In order to successfully apply the photoionization source, it is imperative to know the spectrum of reactant ions and the subsequent ionization reactions leading to the detection of analytes. To that end, an ionization chamber based on the photoionization source that reproduces the ionization processes in the ion mobility spectrometer and facilitates efficient transfer of the product ions into a mass spectrometer was developed. Photoionization of pure gasses and gas mixtures containing air, N2 , CO2 and N2 O and the dopant CH2 Cl2 is discussed. The main product ions of photoionization are identified and compared with the spectrum of reactant ions formed by radioactive and corona discharge sources on the basis of literature data. The results suggest that photoionization by soft X-radiation in the negative mode is more selective than the other sources. In air, adduct ions of O2- with H2 O and CO2 were exclusively detected. Traces of CO2 impact the formation of adduct ions of O2- and Cl- (upon addition of dopant) and are capable of suppressing them almost completely at high CO2 concentrations. Additionally, the ionization products of four alkyl nitrates (ethylene glycol dinitrate, nitroglycerin, erythritol tetranitrate and pentaerythritol tetranitrate) formed by atmospheric pressure chemical ionization induced by X-ray photoionization in different gasses (air, N2 and N2 O) and dopants (CH2 Cl2 , C2 H5 Br and CH3 I) are investigated. The experimental studies are complemented by density functional theory calculations of the most important adduct ions of the alkyl nitrates (M) used for their spectrometric identification. In addition to the adduct ions [M + NO3 ]- and [M + Cl]- , adduct ions such as [M + N2 O2 ]- , [M + Br]- and [M + I]- were detected, and their gas-phase structures and energetics are investigated by density functional theory calculations. Copyright © 2016 John Wiley & Sons, Ltd.

10.
Anal Chim Acta ; 887: 209-215, 2015 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-26320804

RESUMO

L-selectin is a protein with potential importance for numerous diseases and clinical disorders. In this paper, we present a new aptamer-based luminescent assay developed to detect L-selectin. The sensing system working principle is based on Förster Resonance Energy Transfer (FRET) from a donor terbium complex (TbC) to an acceptor cyanine dye (Cy5). In the present approach, the biotinylated aptamer is combined with Cy5-labelled streptavidin (Cy5-Strep) to yield an aptamer-based acceptor construct (Apta-Cy5-Strep), while L-selectin is conjugated using luminescent TbC. Upon aptamer binding to the TbC-labelled L-selectin (L-selectin-TbC), permanent donor-acceptor proximity is established which allows for radiationless energy transfer to occur. However, when unlabelled L-selectin is added, it competes with the L-selectin-TbC and the FRET signal decreases as the L-selectin concentration increases. FRET from the TbC to Cy5 was observed with time-gated time-resolved luminescence spectroscopy. A significant change in the corrected luminescence signal was observed in the dynamic range of 10-500 ng/mL L-selectin, the concentration range relevant for accelerated cognitive decline of Alzheimer's disease, with a limit of detection (LOD) equal to 10 ng/mL. The aptasensor-based assay is homogeneous and can be realized within one hour. Therefore, this method has the potential to become an alternative to tedious heterogeneous analytical methods, e.g. based on enzyme-linked immunosorbent assay (ELISA).


Assuntos
Aptâmeros de Nucleotídeos/química , Transferência Ressonante de Energia de Fluorescência/métodos , Selectina L/análise , Carbocianinas/química , Corantes Fluorescentes/química , Humanos , Selectina L/sangue , Limite de Detecção , Medições Luminescentes/métodos , Térbio/química
11.
Eur J Mass Spectrom (Chichester) ; 21(3): 391-402, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26307720

RESUMO

The application of electrospray ionization (ESI) ion mobility (IM) spectrometry on the detection end of a high-performance liquid chromatograph has been a subject of study for some time. So far, this method has been limited to low flow rates or has required splitting of the liquid flow. This work presents a novel concept of an ESI source facilitating the stable operation of the spectrometer at flow rates between 10 µL mn(-1) and 1500 µL min(-1) without flow splitting, advancing the T-cylinder design developed by Kurnin and co-workers. Flow rates eight times faster than previously reported were achieved because of a more efficient dispersion of the liquid at increased electrospray voltages combined with nebulization by a sheath gas. Imaging revealed the spray operation to be in a rotationally symmetric multijet mode. The novel ESI-IM spectrometer tolerates high water contents (≤90%) and electrolyte concentrations up to 10mM, meeting another condition required of high-performance liquid chromatography (HPLC) detectors. Limits of detection of 50 nM for promazine in the positive mode and 1 µM for 1,3-dinitrobenzene in the negative mode were established. Three mixtures of reduced complexity (five surfactants, four neuroleptics, and two isomers) were separated in the millisecond regime in stand-alone operation of the spectrometer. Separations of two more complex mixtures (five neuroleptics and 13 pesticides) demonstrate the application of the spectrometer as an HPLC detector. The examples illustrate the advantages of the spectrometer over the established diode array detector, in terms of additional IM separation of substances not fully separated in the retention time domain as well as identification of substances based on their characteristic Ims.

12.
Phys Chem Chem Phys ; 17(3): 1740-5, 2015 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-25464013

RESUMO

Hemolysis, the rupturing of red blood cells, can result from numerous medical conditions (in vivo) or occur after collecting blood specimen or extracting plasma and serum out of whole blood (in vitro). In clinical laboratory practice, hemolysis can be a serious problem due to its potential to bias detection of various analytes or biomarkers. Here we present the first "mix-and-measure" method to assess the degree of hemolysis in biosamples using luminescence spectroscopy. Luminescent terbium complexes (LTC) were studied in the presence of free hemoglobin (Hb) as indicators for hemolysis in TRIS-buffer, and in fresh human plasma with absorption, excitation and emission measurements. Our findings indicate dynamic as well as resonance energy transfer (FRET) between the LTC and the porphyrin ligand of hemoglobin. This transfer leads to a decrease in luminescence intensity and decay time even at nanomolar hemoglobin concentrations either in buffer or plasma. Luminescent terbium complexes are very sensitive to free hemoglobin in buffer and blood plasma. Due to the instant change in luminescence properties of the LTC in presence of Hb it is possible to access the concentration of hemoglobin via spectroscopic methods without incubation time or further treatment of the sample thus enabling a rapid and sensitive detection of hemolysis in clinical diagnostics.


Assuntos
Análise Química do Sangue/métodos , Complexos de Coordenação/química , Hemoglobinas/análise , Soro/química , Térbio/química , Hemoglobinas/química , Humanos , Luminescência
13.
Appl Spectrosc ; 68(9): 1030-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25226257

RESUMO

In this paper the concept of a compact high-resolution spectrometer based on the combination of dispersive and interferometric elements is presented. Dispersive elements are used to spectrally resolve the light in one direction with coarse resolution (Δλ < 0.5 nm), while perpendicular to that direction an etalon provides high spectral resolution (Δλ < 50 pm). This concept for two-dimensional spectroscopy has been implemented for the wavelength range λ = 350-650 nm. Appropriate algorithms for reconstructing spectra from the two-dimensional raw data and for wavelength calibration were established in an analysis software. Potential applications for this new spectrometer are Raman and laser-induced breakdown spectroscopy (LIBS). Resolutions down to 28 pm (routinely 54 pm) could be realized for these applications.

14.
J Biomed Opt ; 19(10): 101506, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24989900

RESUMO

Nanobioconjugates have been synthesized using cadmium selenide quantum dots (QDs), europium complexes (EuCs), and biotin. In those conjugates, long-lived photoluminescence (PL) is provided by the europium complexes, which efficiently transfer energy via Förster resonance energy transfer (FRET) to the QDs in close spatial proximity. As a result, the conjugates have a PL emission spectrum characteristic for QDs combined with the long PL decay time characteristic for EuCs. The nanobioconjugates synthesis strategy and photophysical properties are described as well as their performance in a time-resolved streptavidin-biotin PL assay. In order to prepare the QD-EuC-biotin conjugates, first an amphiphilic polymer has been functionalized with the EuC and biotin. Then, the polymer has been brought onto the surface of the QDs (either QD655 or QD705) to provide functionality and to make the QDs water dispersible. Due to a short distance between EuC and QD, an efficient FRET can be observed. Additionally, the QD-EuC-biotin conjugates' functionality has been demonstratedin a PL assay yielding good signal discrimination, both from autofluorescence and directly excited QDs. These newly designed QD-EuC-biotin conjugates expand the class of highly sensitive tools for bioanalytical optical detection methods for diagnostic and imaging applications.


Assuntos
Técnicas Biossensoriais/métodos , Biotina/imunologia , Európio/química , Imunoensaio/métodos , Medições Luminescentes/métodos , Pontos Quânticos , Estreptavidina/imunologia , Corantes Fluorescentes/química , Nanoconjugados
15.
Phys Chem Chem Phys ; 16(13): 6060-7, 2014 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-24556813

RESUMO

A new functional luminescent lanthanide complex (LLC) has been synthesized with terbium as a central lanthanide ion and biotin as a functional moiety. Unlike in typical lanthanide complexes assembled via carboxylic moieties, in the presented complex, four phosphate groups are chelating the central lanthanide ion. This special chemical assembly enhances the complex stability in phosphate buffers conventionally used in biochemistry. The complex synthesis strategy and photophysical properties are described as well as the performance in time-resolved Förster Resonance Energy Transfer (FRET) assays. In those assays, this biotin-LLC transferred energy either to acceptor organic dyes (Cy5 or AF680) labelled on streptavidin or to quantum dots (QD655 or QD705) surface-functionalised with streptavidins. The permanent spatial donor-acceptor proximity is assured through strong and stable biotin-streptavidin binding. The energy transfer is evidenced from the quenching observed in donor emission and from a decrease in donor luminescence decay, both associated with simultaneous increase in acceptor intensity and in the decay time. The dye-based assays are realised in TRIS and in PBS, whereas QD-based systems are studied in borate buffer. The delayed emission analysis allows for quantifying the recognition process and for auto-fluorescence-free detection, which is particularly relevant for application in bioanalysis. In accordance with Förster theory, Förster-radii (R0) were found to be around 60 Å for organic dyes and around 105 Å for QDs. The FRET efficiency (η) reached 80% and 25% for dye and QD acceptors, respectively. Physical donor-acceptor distances (r) have been determined in the range 45-60 Å for organic dye acceptors, while for acceptor QDs between 120 Å and 145 Å. This newly synthesised biotin-LLC extends the class of highly sensitive analytical tools to be applied in the bioanalytical methods such as time-resolved fluoroimmunoassays (TR-FIA), luminescent imaging and biosensing.


Assuntos
Complexos de Coordenação/química , Transferência Ressonante de Energia de Fluorescência , Térbio/química , Biotina/química , Biotina/metabolismo , Fluorimunoensaio , Pontos Quânticos/química , Estreptavidina/química , Estreptavidina/metabolismo
16.
J Chem Phys ; 140(4): 044907, 2014 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-25669583

RESUMO

In this paper, we investigate interactions and phase transitions in polyelectrolyte-surfactant complexes formed between a cationic azobenzene-containing surfactant and two types of polyelectrolytes: natural (DNA) or synthetic (PAA: poly acrylic acid). The construction of a phase diagram allowed distancing between four major phases: extended coil conformation, colloidally stable compacted globules, colloidal instability range, and surfactant-stabilized compact state. Investigation on the complexes' properties in different phases and under irradiation with UV light provides information about the role of the surfactant's hydrophobic trans isomers both in the formation and destruction of DNA and PAA globules as well as in their colloidal stabilization. The trans isomer shows much stronger affinity to the polyelectrolytes than the hydrophilic cis counterpart. There is no need for complete compensation of the polyelectrolyte charges to reach the complete compaction. On contrary to the findings previously reported in the literature, we demonstrate - for the first time - complete polyelectrolyte compaction which occurs already at 20% of DNA (and at 50% of PAA) charge compensation. The trans isomer plays the main role in the compaction. The aggregation between azobenzene units in the photosensitive surfactant is a driving force of this process. The decompaction can be realized during UV light irradiation and is strongly influenced by the interplay between surfactant-surfactant and surfactant-DNA interactions in the compacted globules.


Assuntos
Resinas Acrílicas/química , DNA/química , Tensoativos/química , Animais , Compostos Azo/química , Bovinos , Hidrodinâmica , Interações Hidrofóbicas e Hidrofílicas , Isomerismo , Modelos Químicos , Timo , Raios Ultravioleta
17.
Biosens Bioelectron ; 52: 288-92, 2014 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-24071364

RESUMO

Novel luminescent ratiometric nanosensors (QD-NAPTHs) were prepared based on cadmium telluride (CdTe655) quantum dots as luminescent nanoscaffolds with naphthyridine dyes as fluorescent receptors. This biosensing bifluorophoric nanosystem has been designed to achieve detection of guanosine 3',5'-cyclic monophosphate (cyclic GMP) in buffered media. Cyclic GMP is a secondary messenger that is an important factor for detecting cancer, diabetes and, cardiovascular diseases. Due to low concentration levels, even in pathological conditions, sensitive cGMP detection remains a challenge for modern biomedical diagnostics. Here, QD-NAPTH nanosensors were tested in the presence of a target nucleotide and with various structural cGMP analogues. Steady-state fluorescence spectroscopy was used to monitor a change in the nucleotide concentration. A 5-fold increase in naphthyridine fluorescence with a simultaneous decrease in QD luminescence was observed after adding 50 µM of cGMP. Using this novel nanosystem with ratiometric detection, it was possible to recognize cGMP with limit of detection (3σ) equal to 70 ng/ml. Moreover, the enhancement in fluorescence upon interaction with the target nucleotide constitutes a favourable approach towards the detection of cGMP in buffered media. These bifluorophoric nanosensors have a potential for application in fluorescence microscopy imaging and in-vitro assays.


Assuntos
Técnicas Biossensoriais , Compostos de Cádmio/química , GMP Cíclico/isolamento & purificação , Telúrio/química , GMP Cíclico/química , Nanopartículas/química , Naftiridinas/química
18.
Anal Bioanal Chem ; 405(26): 8525-37, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23975087

RESUMO

Calcium (Ca(2+)) is a ubiquitous intracellular second messenger and involved in a plethora of cellular processes. Thus, quantification of the intracellular Ca(2+) concentration ([Ca(2+)]i) and of its dynamics is required for a comprehensive understanding of physiological processes and potential dysfunctions. A powerful approach for studying [Ca(2+)]i is the use of fluorescent Ca(2+) indicators. In addition to the fluorescence intensity as a common recording parameter, the fluorescence lifetime imaging microscopy (FLIM) technique provides access to the fluorescence decay time of the indicator dye. The nanosecond lifetime is mostly independent of variations in dye concentration, allowing more reliable quantification of ion concentrations in biological preparations. In this study, the feasibility of the fluorescent Ca(2+) indicator Oregon Green Bapta-1 (OGB-1) for two-photon fluorescence lifetime imaging microscopy (2P-FLIM) was evaluated. In aqueous solution, OGB-1 displayed a Ca(2+)-dependent biexponential fluorescence decay behaviour, indicating the presence of a Ca(2+)-free and Ca(2+)-bound dye form. After sufficient dye loading into living cells, an in situ calibration procedure has also unravelled the Ca(2+)-free and Ca(2+)-bound dye forms from a global biexponential fluorescence decay analysis, although the dye's Ca(2+) sensitivity is reduced. Nevertheless, quantitative [Ca(2+)]i recordings and its stimulus-induced changes in salivary gland cells could be performed successfully. These results suggest that OGB-1 is suitable for 2P-FLIM measurements, which can gain access to cellular physiology.


Assuntos
Compostos de Anilina/análise , Cálcio/análise , Fluoresceínas/análise , Corantes Fluorescentes/análise , Microscopia de Fluorescência/métodos , Imagem Óptica/métodos , Animais , Baratas/citologia , Glândulas Salivares/citologia
19.
Anal Bioanal Chem ; 405(22): 7031-9, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23887276

RESUMO

The detection of hydrogen sulfide (H2S) by 2 + 1 resonance-enhanced multi-photon ionization (REMPI) and the application of H2S as a laser dopant for the detection of polar compounds in laser ion mobility (IM) spectrometry at atmospheric pressure were investigated. Underlying ionization mechanisms were elucidated by additional studies employing a drift cell interfaced to a time-of-flight mass spectrometer. Depending on the pressure, the primary ions H2S(+), HS(+), S(+), and secondary ions, such as H3S(+), were observed. The 2 + 1 REMPI spectrum of H2S near λ = 302.5 nm was recorded at atmospheric pressure. Furthermore, the limit of detection and the linear range were established. In the second part of the work, H2S was investigated as an H2O analogous laser dopant for the ionization of polar substances by proton transfer. H2S exhibits a proton affinity (PA) similar to that of H2O, but a significantly lower ionization energy facilitating laser ionization. Ion-molecule reactions (IMR) of H3S(+) with a variety of polar substances with PA between 754.6 and 841.6 kJ/mol were investigated. Representatives of different compound classes, including alcohols, ketones, esters, and nitroaromatics were analyzed. The IM spectra resulting from IMR of H3S(+) and H3O(+) with these substances are similar in structure, i.e., protonated monomer and dimer ion peaks are found depending on the analyte concentration.


Assuntos
Sulfeto de Hidrogênio/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Pressão Atmosférica , Desenho de Equipamento , Íons/análise , Lasers , Limite de Detecção , Prótons , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentação
20.
Sensors (Basel) ; 13(6): 7170-83, 2013 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-23727953

RESUMO

In many biological and environmental applications spatially resolved sensing of molecular oxygen is desirable. A powerful tool for distributed measurements is optical time domain reflectometry (OTDR) which is often used in the field of telecommunications. We combine this technique with a novel optical oxygen sensor dye, triangular-[4] phenylene (TP), immobilized in a polymer matrix. The TP luminescence decay time is 86 ns. The short decay time of the sensor dye is suitable to achieve a spatial resolution of some meters. In this paper we present the development and characterization of a reflectometer in the UV range of the electromagnetic spectrum as well as optical oxygen sensing with different fiber arrangements.


Assuntos
Oxigênio/análise , Espectrofotometria Ultravioleta , Corantes/química , Tecnologia de Fibra Óptica , Polímeros/química
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA