New immobilization method for SAW-biosensors: covalent attachment of antibodies via CNBr.

Surface acoustic wave devices (SAWs) based on horizontally polarized shear waves can be used as mass-sensitive immunosensors. If the standard material, aluminium, is used for the interdigital transducers (IDTs) of these devices a pretreatment with a shielding layer of polyimide is necessary to prevent corrosion processes. However, this layer also prevents an easy covalent attachment of the necessary receptive molecules. Therefore, we developed a new activation method of this inert polyimide surface by treating with cyanogenbromide using the established cyano-transfer-technique. With a subsequent covalent attachment of anti-glucose oxidase (anti-GOD) a very sensitive SAW-immunosensor for glucose oxidase (GOD) can be obtained.

Covalent photolinker-mediated immobilization of an intermediate dextran layer to polymer-coated surfaces for biosensing applications.

A new method is presented for the covalent binding of dextran as an intermediate layer on surface acoustic wave (SAW) devices. For biosensing applications in aqueous media commercially available SAW devices require surface passivation to prevent corrosion of the aluminum device structures in electrolytes. Thin films of polyimide and parylene revealed exceptional passivation properties. They were used as a base for dextran immobilization. Covalent binding of dextran to polymer-coated surfaces was achieved by photoimmobilization. Aryldiazirine-functionalized bovine serum albumin served as the multifunctional light-activable linking agent (photolinker polymer). Dextran and photolinker polymer were mixed and photobonded to sensor surfaces. Essential photoimmobilization parameters were optimized. The binding of proteins to dextran applying carbodiimide chemistries was exemplified with antiurease antibodies and the feasibility of specific immunosensing was investigated on SAW sensors connected to a fluid handling system.

Determination of ascorbic acid in nanolitre samples by means of capillary batch injection analysis.

Capillary batch injection analysis (CBIA) is applied to the determination of ascorbic acid in nanolitre samples at neutral pH. It is demonstrated that pulsed amperometric detection is superior to constant-potential amperometric detection with respect to signal stability. The calibration plot is linear within the studied concentration range between 0.5 mM and 5 mM and the reproducibility is characterized by a relative standard deviation of 0.80% for 10 successive injections of 188 nL of 3.5 mM ascorbic acid. The attainable sample throughput is up to 500 per hour.

Chemical microanalytical systems: objectives and latest developments.

Chemical microanalysis systems are at a very early state of development. At the present time a modular approach with subsystems carrying out the various functions such as sampling, preparation and measurement seems to be favored by the developers. The marketpotential of the systems can be seen primarily in the area of inexpensive monitoring equipment for environmental and process control. Reduction of size leads also to smaller sample volumes, less waste and shorter analysis time: Miniaturization is in many cases an essential feature for biological and in-vivo medical investigations, while some properties inherently associated with miniaturization can also lead to substantial improvements in the technical performance of the microsystem as compared to its macro version. This will be demonstrated by discussing some microsystems which include spectroscopic detection systems:- Miniature separation instruments coupled with advanced miniaturized detection device based on photothermal spectroscopy- Integrated-optical NIR evanescent wave sensor systems- Optochemical and biosensor systems

Optimisation of an integrated optical evanescent wave absorbance sensor for the determination of chlorinated hydrocarbons in water.

The suitability of an integrated optical chemical sensor for the determination of highly volatile chlorinated hydrocarbons in aqueous solutions has been proven. The analytes are detected by NIR absorption spectrometry in the evanescent field of an integrated optical strip waveguide generated in a BGG31 (Schott, Germany) glass substrate, which is coated with a hydrophobic polymer superstrate as sensing layer. It has been shown that the sensitivity increases when the refractive index of the superstrate is increased from 1.333 up to 1.46. Different UV-cured polysiloxanes with low cross sensitivity to water have been prepared. Due to the good light transmission properties of the IO-sensors prepared by this method, quantitative measurements have been performed with the model system trichloroethene (TCE) in water. A detection limit of 22 ppm has been found and the sensor response times (t(90)-value) are between five and fourteen minutes for a coating thickness of around 30 microm. The sensor response is totally reversible. The analyte desorbes in air within 2 min. The enrichment of trichloroethene in the polysiloxane coating can be described by film diffusion through the aqueous boundary layer as rate determining step.

Determination of hydrocarbons in water by evanescent wave absorption spectroscopy in the near-infrared region.

An home-made EFA (Evanescent Field Absorbance)-sensor has been tested for the determination of hydrocarbons in water. The investigations have been performed both with crude oil emulsions and petrol solutions. Cuvette and evanescent wave spectra of crude oil and petrol in the near-infrared region are presented and discussed. The concentration of aromatic compounds in crude oil can be determined semiquantitatively by the standard addition method. The sorption behaviour of the hydrocarbons in the cladding of the fiberoptic sensor has been investigated and a correlation between the sensor signal and the concentration of the aqueous hydrocarbon emulsion/solution could be shown. The desorption of the enriched molecules after the measurements is also presented. The petrol molecules evaporate in ambient air so that the sensor is easily regenerated. In case of oil measurements the hydrocarbon molecules cannot be removed by rinsing the sensor with clear water or by evaporating them in ambient air. It has to be regenerated by washing it with a high volatile solvent instead.

ISFETs with sputtered sodium alumino-silicate glass membranes.

Sodium ion sensitive field-effect transistors (ISFETs) based on reactively sputtered sodium aluminosilicate (NAS) glass membranes are investigated. Using an inverted cylindrical magnetron, NAS films with defined and reproducible composition are obtained. Radiation damage in the gate dielectric is largely eliminated by annealing at 450 degrees C. The Na(+) sensitivity, the selectivity with respect to H(+) and K(+) ions, the response time, the conditioning period, the long-term drift, and the lifetime are evaluated. The ISFETs exhibit a near-Nernstian Na(+) response down to 10(-4) mol/l in solutions of pH>/=7. The NAS films are physically very robust and have excellent chemical durability, e.g., a sensor with a 110 nm thick membrane lasts for about 8 months. The overall sensor properties compare well with those of a commercial Na(+) selective glass electrode.

Trace analysis of pollutants in water using the photothermal interferometry as HPLC detector.

A new procedure including high performance liquid chromatography in combination with photothermal interference spectroscopy as detection device (HPLC/PIS) has been proposed, optimized and its figures of merit for pesticide residue analysis are shown. The flowing sample under study is set in one arm of a Mach-Zehnder interferometer, and its refractive index is modulated by a periodically chopped continuous wave argon ion laser. As chopper, an acousto optical modulator has been introduced to switch the excitation laser beam between different lines (457 nm, 488 nm, 514 nm) simultaneously. Thus a multi component analysis can be realized either by using an HPLC-system in front of the PIS device or by a multi line Ar(+)-laser, directly. The limit of detection of the HPLC/PIS system reached 71 microg/l of the pesticide di-nitro-ortho-cresol (DNOC).

Integrated optical NIR-evanescent wave absorbance sensorfor chemical analysis.

A new, long-path integrated optical (IO) sensor for the detection of non-polar organic substances is described. The sensing layer deposited on a planar multimode IO structure is built by a suitable silicone polymer with lower refractive index (RI). It acts as a hydrophobic matrix for the reversible enrichment of non-polar organic contaminants from water or air. Light from the near-infrared (NIR) range is coupled into the planar structure and the evanescent wave part of the light field penetrating into the silicone layer interacts with the enriched organic species. As a result, light is absorbed at the characteristic frequencies of the corresponding C-H, N-H or O-H overtone and combination band vibrations of the analytes. To perform evanescent field absorbance (EFA) measurements, the arc-shaped strip waveguide structure of 172 mm interaction length was adapted to a tungsten-halogen lamp and an InGaAs diode array spectrograph over gradient index fibers. Dimethyl-co-methly(phenyl)polysiloxanes with varying degrees of phenylation were prepared and used as sensitive coating materials for the IO structure. Light attenuation in the arc-shaped waveguides is high and typical insertion losses in the range of 14-18 dB were obtained. When the coated sensors were brought in contact with aqueous samples, the light transmission decreases, which is due to the formation of H(2)O micro-emulsions in the silicone superstrates. Nevertheless, after reaching constant light transmissions, absorbance spectra of aqueous trichloroethene samples were successfully collected. For gas measurements, where water cross sensitivity problems are absent, the sensitivity of the IO device for trichloroethene was tested as a function of the RI of the silicone superstrate. The slope of the TCE calibration function increases by a factor of 10 by using a poly(methylphenylsiloxane) layer with a RI of 1.449 instead of poly(dimethylsiloxane) (RI: 1.41). A comparison of the IO-EFA and an earlier developed fiber-optic EFA sensor for trichloroethene measurements in the gas phase showed an increase in sensitivity per unit length of the waveguide by a factor of up to 120.

Chlorophyll fluorescence biosensor for the detection of herbicides.

A biosensor is described for the detection of triazine and phenylurea herbicides in drinking water by kinetic measurements of endogenous chlorophyll fluorescence in isolated chloroplasts from higher plants. The pocket-size device uses a diode laser for simultaneous excitation of sample and reference channels, and photodiodes for detection of the emitted light. The biological material can be supplied as a freeze-dried powder, stable for 21 days at room temperature, for 6 weeks at 4 ( degrees )C or for at least 9 months at -20 ( degrees )C. The detection of 0.1 microg/l of a single herbicide, as required by European Community legislation on drinking water quality, can be achieved without prior extraction of the sample.

Synthesis of C-11 iodoantipyrine for positron emission tomography.

We have developed a method for the synthesis of C-11 iodoantipyrine. Carbon-11-labeled methyl iodide, prepared from 11CO2, was used to methylate 3-methyl-1-phenyl-2-pyrazolin-5-one to form C-11 antipyrine. Following silica-gel column chromatography and iodination, radiochemical purity of the C-11 iodoantipyrine was more than 99.5%, with a 10% yield and a specific activity of 30 mCi/mumol. Preliminary animal studies showed complete cerebral extraction and local cerebral blood-flow values that were within 4.6% of those obtained using C-14 iodoantipyrine. The C-11 analog, with positron emission tomography, will facilitate local cerebral blood-flow studies in human subjects.