Surface plasmon resonance biosensor for determination of tetrodotoxin: prevalidation study

A label-free surface plasmon resonance biosensor method was applied to determine tetrodotoxin (TTX) in pufferfish matrixes using an antibody inhibition assay format. A prevalidation study was conducted to demonstrate the assay performance characteristics, such as selectivity, LOD, LOQ, repeatability, reproducibility, and accuracy. Three participating laboratories reported standard curves in buffer and pufferfish matrix. A set of blind samples with TTX spiked into buffer as well as in 10% pufferfish extract were analyzed. Additionally, three blind naturally contaminated samples were analyzed, and the results were compared to those obtained using a reference method (HPLC/electrospray ionization-selected reaction monitoring-MS). The developed method was demonstrated to be capable of detecting TTX in pufferfish matrix standard samples in a broad concentration range (2-9000 ng/mL) with an LOD of 1.5 ng/mL. Between-laboratory recovery values were in the range of 51-190% with a mean of 107%, and 64-180% with a mean of 103% for TTX-spiked samples in buffer and pufferfish matrix, respectively. Between-laboratory recoveries were in the satisfactory range of 101-119% for naturally contaminated samples. This robust, rapid, and noninvasive method may serve as an attractive alternative to established methods for detection of TTX in pufferfish extracts.     

Tail-labelling of DNA probes using modified deoxynucleotide triphosphates and terminal deoxynucleotidyl transferase. Application in electrochemical DNA hybridization and protein-DNA binding assays

A simple approach to DNA tail-labelling using terminal deoxynucleotidyl transferase and modified deoxynucleoside triphosphates is presented. Amino- and nitrophenyl-modified dNTPs were found to be good substrates for this enzyme giving 3'-end stretches of different lengths depending on the nucleotide and concentration. 3-Nitrophenyl-7-deazaG was selected as the most useful label because its dNTP was efficiently incorporated by the transferase to form long tail-labels at any oligonucleotide. Accumulation of many nitrophenyl tags per oligonucleotide resulted in a considerable enhancement of voltammetric signals due to the nitro group reduction, thus improving the sensitivity of electrochemical detection of the tail-labelled probes. We demonstrate a perfect discrimination between complementary and non-complementary target DNAs sequences by tail-labelled hybridization probes as well as the ability of tumour suppressor p53 protein to recognize a specific binding site within tail-labelled DNA substrates, making the methodology useful in electrochemical DNA hybridization and DNA-protein interaction assays.     

Bi(Pb)–Sr–Ca–Cu–O layers prepared by a new method of laser ablation in air

Bi(Pb)–Sr–Ca–Cu–O layers were prepared by laser ablation in air, that is without using a vacuum chamber. At this method a much higher density of pulse energy of laser beam is required (10–30 J cm^–2) in comparison with the standard method (1–3 J cm^–2). The mentioned condition has specific effects on the nature of the transport of material from the target. As follows from the electron microprobe analysis of the layers, the starting composition of the transported material is identical with that of the superconducting phase sintered in the target; one exception represents Pb, the content of which in the transported material is lower. The image of the process is complemented by UV/VIS spectroscopy scans of the plasma plume.     

Zur Synthese von 1-(3,3-Dimethyl-2-norbornyl)-äthanon

An improved synthesis of the title compound (1) is described. The catalytical action of variousLewis acids on the sterically hinderedDiels-Alder reaction of mesityl oxide with cyclopentadiene has been investigated. The mixture ofendo- andexo-isomers of the unsaturated intermediate2 yielded pure1 on hydrogenation and isomerization by sodium methoxide. The proof of theexo-configuration of the acetyl group has been achieved by mass spectra and 100 MHz¹H-NMR spectra.

---

---

Herrn Prof. Dr.M. Pailer mit den besten Wünschen zum 65. Geburtstag gewidmet.     

Elektrothermische atomisierung von quecksilber bei anwesenheit von thiolhaltigen verbindungen

Electrothermal atomisation of mercury in the presence of thiols. Sample stabilisation during the drying step is important for the determination of mercury by electrothermal a.a.s. Addition of complexing agents with thiol groups makes possible the analysis of pure aqueous solutions. Mercury can be determined at concentrations down to 5 × 10^-6% with a reproducibility of 2–10%. Small amounts of acids and salts depress the efficiency of stabilisation, so that separation becomes necessary. The use of a selective ion-exchanger with thiol groups solves the problems of sample storage, separation and stabilisation. The enrichment enhances the sensitivity by two orders of magnitude. Determinations of 2–20 ppb Hg in water, 0.2–2 M HC1 and 0.2–1 M NaCl are reported.     

Study of the stability of pyrimidine series cytostatics,ftorafur and fluorouracil injections

The stability of the cytostatics, ftorafur and fluorouracil injections, was studied in dependence on irradiation with UV light and on the medium temperature and pH. In the first stage of the thermal and photochemical decomposition of ftorafur, tetrahydrofuran dissociated from position N₁ in the pyrimidine ring and 5-fluorouracil was formed. In the second stage of the thermal and photochemical decomposition of ftorafur, the pyrimidine ring opened between N₃ and C₄ and between C₆ and N₁, with formation of urea. Ftorafur, fluorouracil, and urea were determined using thin-layer chromatography and spectrophotometry in the UV and visible region.     

Affinity liquid chromatography and capillary electrophoresis of seminal plasma proteins

Interactions of boar, bull, and human seminal plasma proteins with heparin and phosphorylcholine were studied by affinity LC using heparin immobilized to a Toyopearl support. A step gradient elution from 0.15 to 1.50 M NaCl was employed to elute the seminal plasma proteins. Relative amounts of the heparin-binding fraction of seminal plasma proteins (H+) in seminal plasma of three species were determined. Further on, the fraction of seminal plasma proteins interacting with phosphorylcholine-binding proteins (P+) was evaluated. P+ proteins were not found in human seminal plasma and their highest amount was present in bull seminal plasma. A CE method was developed for separation of seminal plasma proteins. Various capillaries and separation conditions were tested; the best resolution was obtained in a bare-silica capillary, with a micellar system consisting of a 0.02 M borate buffer and 0.05 M SDS pH 10.0. The optimized conditions were applied to the identification of the components in boar plasma.     

Electronic, infrared, and 1HNMR spectral studies of the novel charge-transfer complexes of o-tolidine and p-toluidine with alternation pi-acceptors (3,5-dinitro benzoic acid and 2,6-dichloroquinone-4-chloroimide) in CHCl3 solvent

The rapid interaction between o-tolidine and p-toluidine (pi-donors) with the pi-acceptors, e.g., 3,5-dinitrobenzoic acid (DNB) and 2,6-dichloroquinone-4-chloroimide (DCQ) results in the formation of 1:1 charge-transfer complexes as the final products, [(o-tolidine) (acceptor)] and [(p-toluidine) (acceptor)]. The final products of the reactions have been isolated and characterized using FTIR, 1HNMR spectroscopy and elemental analysis as well as photometric titration. The stoichiometry and apparent formation constants of the complexes formed were determined by applying the conventional spectrophotometric molar ratio method.     

Vapour permeation and sorption in fluoropolymer gel membrane based on ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoromethylsulphonyl)imide

The emissions of hydrocarbons from fossil fuels into atmosphere entail both an economic loss and an environmental pollution. Membrane separations can be used for vapour recovery and/or vapour removal from the permanent gas stream, given that the appropriate membrane is identified. A neat poly(vinylidene fluoride-co-hexafluoropropylene) membrane is impermeable to both the representatives of aliphatic hydrocarbons and branched hydrocarbons, namely hexane and isooctane, whereas the permeation flux is enhanced by the presence of 80 mass % of the ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoromethylsulphonyl)imide in the membrane, as detailed in this work. The permeabilities of hydrocarbon vapours were determined from the binary mixture containing hydrocarbon and nitrogen to simulate the real input of an air stream containing a condensable hydrocarbon. The diffusion coefficient determined from sorption measurements was higher for hexane, as would be expected for a smaller molecule, whereas both the sorption isotherms and permeabilities of the hydrocarbons studied were found to be almost identical. It is possible that the sorption effect predominates in the transport mechanism for VOCs/N₂ separations.