In this work, the coupling of liquid nanochromatography to NanoFlow electrospray mass spectrometry was evaluated for the detection of DNA adducts. The NanoFlow ES LC/MS system was compared with the capillary and conventional ES LC/MS system by analyzing an in vitro reaction mixture resulting from the interaction of 2'-deoxyguanosine 5'-monophosphate with bisphenol A diglycidyl ether and by injecting 2'-deoxyadenosine. By using NanoFlow ES LC/MS, the mass sensitivity could be improved by a factor of 3300. Three different injection methods used in liquid nanochromatography, i.e., split, large-volume, and column-switching injections were compared in terms of sensitivity. Furthermore, NanoFlow ES LC/MS was used to detect 2'-deoxynucleotide adducts isolated from an in vitro mixture of calf thymus DNA and bisphenol A diglycidyl ether. Different 2'-deoxynucleotide adducts could be identified by monitoring typical product ions, diagnostic for the adducts.
ABSTRACT:We report here on a new potentiometric biosensing principle for the detection of antibody−antigen interactions at the sensing membrane surface without the need to add a label or a reporter ion to the sample solution. This is accomplished by establishing a steady-state outward flux of a marker ion from the membrane into the contacting solution. The immunobinding event at the sensing surface retards the marker ion, which results in its accumulation at the membrane surface and hence in a potential response. The ion-selective membranes were surface-modified with an antibody against respiratory syncytial virus using click chemistry between biotin molecules functionalized with a triple bond and an azide group on the modified poly (vinyl chloride) group of the membrane. The bioassay sensor was then built up with streptavidin and subsequent biotinylated antibody. A quaternary ammonium ion served as the marker ion. The observed potential was found to be modulated by the presence of respiratory syncytial virus bound on the membrane surface. The sensing architecture was confirmed with quartz crystal microbalance studies, and stir effects confirmed the kinetic nature of the marker release from the membrane. The sensitivity of the model sensor was compared to that of a commercially available point-of-care test, with promising results.O
Capillary electrophoresis was coupled successfully and reliably to potentiometric sensors, which are based on an ionically conductive rubber phase coating, applied on a 250 microm diameter metal substrate. The membrane components included potassium tetrakis(p-chlorophenyl)borate (TCPB), bis(2-ethylhexyl)sebacate (DOS), and high molecular mass poly(vinyl chloride) (PVC). Potentiometry reveals a very sensitive CE detection mode, with sub-micromolar detection limits for amines and the randomly chosen drugs quinine, clozapine, cocaine, heroine, noscapine, papaverine, and ritodrine. The lowest detection limit, 1 x 10(-8) M injected concentration, was obtained for the quaternary ammonium compound tetrahexylammonium chloride. The more polar lower aliphatic amines and the biogenic amines dopamine, adrenaline, and cadaverine have much higher detection limits. The detection limits are log P dependent. Addition of a commercially available calixarene molecule or a synthetic macrocyclic amphiphilic receptor molecule to the electrode coatings enhanced the sensitivity respectively for the lower aliphatic amines and for the biogenic amines. A transpose of the Nikolskii-Eisenman-type function was suggested and used to convert the signal of the detector to a concentration-dependent signal.
A systematic study has been described on the laser diffraction (LD) and static image analysis (SIA) of rectangular particles [1]. To rule out powder sampling, sample dispersion and particle orientation as a possible root cause for differences in size distribution profile, powder samples were initially immobilized by means of a dry disperser onto a glass plate. For a defined region of the glass plate the diffraction pattern as induced by the dispersed particles, and the 2D dimensions of the individual particles were measured by LD and optical microscopy, respectively. Correlation between LD and SIA could be demonstrated considering the scattering intensity of the individual particles as the most dominating factor. For both spherical and rectangular particles, theory explains the latter to relate to the square of their projected area. In traditional LD, the size distribution profile is dominated by the maximum projected area of the particles (A), and the diffraction diameters of a rectangular particle with length L and breadth B are perceived by the LD instrument to correspond by approximation with spheres having a diameter of ∅ L and ∅ B , respectively. Weighting for differences in scattering intensity between spherical and Page 3 of 56 rectangular particles exlains each rectangular particle to contribute to the overall LD volume probability distribution proportional to A 2 /L and A 2 /B. Accordingly, for rectangular particles this scattering intensity weighted diffraction diameter (SIWDD) concept explains an overestimation of their shortest dimension and an underestimation of their longest dimension. For this study various samples have been analysed with the longest dimension of the particles ranging from ca. 10 to 1000 µm. For a variety of pharmaceutical powders all with a different rectangular particle size and shape, the demonstrated correlation between LD and SIA aims to facilitate the user in a better validation of LD methods based on SIA data.
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