Method for confirmation of synthetic corticosteroids in doping urine samples by liquid chromatography-electrospray ionisation mass spectrometry.

In this study, we report on the development of a method to confirm simultaneously nine of the most commonly abused synthetic corticosteroids in urine based on liquid chromatography-electrospray ionisation mass spectrometry. A considerable simplified sample preparation procedure, including liquid-liquid phase extraction with Extrelut-NT3 columns, provided both excellent sample purification and high overall recoveries. Complete HPLC separations were obtained on a reversed-phase column with 1 mM ammonium acetate-acetonitrile (60:40, v/v) as mobile phase. Mass spectral acquisition was done in the negative ion, and selected ion monitoring modes to identify the drugs with at least three characteristic ions. Detection limits were determined at < or =1 ng/ml and the confirmation limits at 1 to 5 ng/ml.

Electrokinetically driven microfluidic chips with surface-modified chambers for heterogeneous immunoassays.

This article presents the first example of a microfluidic chip for heterogeneous bioassays using a locally immobilized biospecific layer and operated electrokinetically. The reaction chamber has picoliter dimensions and is integrated into a network of microchannels etched in glass. The high affinity of protein A (PA) for rabbit immunoglobulin G (rIgG) was exploited for chip testing, with PA being immobilized on microchannel walls and fluorescently labeled (Cy5) rIgG serving as sample. It was possible to operate the chip in an immunoaffinity chromatographic manner, using electrokinetically pumped solutions. Concentration of antibody from dilute solution onto the solid phase was demonstrated, with signal gains of approximately 30 possible. A dose-response curve for Cy5-rIgG was obtained for concentrations down to 50 nM, for an incubation time of 200 s. The flexibility of chip layout was demonstrated for competitive immunoassay of rIgG, using both a combined sample/tracer incubation and sequential addition of these solutions. With assay times generally below 5 min for this unoptimized device, the microfluidic approach described shows great potential for many high-throughput screening applications.

A two-electrode configuration for simplified amperometric detection in a microfabricated electrophoretic separation device.

The simplified amperometric detection scheme demonstrated is based on the amperometric working and electrophoretic ground electrodes only. The latter serves as counter and pseudo-reference as well. It is shown via the successful determination of neurotransmitters, ascorbic acid and phenols on gold or platinum working electrodes that this approach is feasible for detection on a channel based electrophoretic separation device. Also presented is the detection of carbohydrates and amino acids with copper electrodes. The results were found to be similar to those obtained with conventional capillary systems with amperometric detection, albeit at much reduced analysis times.

Integrated capillary electrophoresis devices with an efficient postcolumn reactor in planar quartz and glass chips.

Methods to fabricate planar capillary electrophoresis devices integrated with a postcolumn reactor in fused silica (quartz) and Pyrex glass are presented. Quartz is etched at ∼1 µm/min with a 2.1:1 width-to-depth ratio using a Cr/Au/Cr metal mask and concentrated HF/HNO(3). On-chip postcolumn reaction of o-phthaldialdehyde (OPA) and amino acids gave theoretical plate numbers up to 83 000 and ∼90 ms peak widths, corresponding to 14 plates/V and a 0.5 µm theoretical plate height. The reactor geometry caused only a 10% degradation in efficiency.

Micromachining a miniaturized capillary electrophoresis-based chemical analysis system on a chip.

Micromachining technology was used to prepare chemical analysis systems on glass chips (1 centimeter by 2 centimeters or larger) that utilize electroosmotic pumping to drive fluid flow and electrophoretic separation to distinguish sample components. Capillaries 1 to 10 centimeters long etched in the glass (cross section, 10 micrometers by 30 micrometers) allow for capillary electrophoresis-based separations of amino acids with up to 75,000 theoretical plates in about 15 seconds, and separations of about 600 plates can be effected within 4 seconds. Sample treatment steps within a manifold of intersecting capillaries were demonstrated for a simple sample dilution process. Manipulation of the applied voltages controlled the directions of fluid flow within the manifold. The principles demonstrated in this study can be used to develop a miniaturized system for sample handling and separation with no moving parts.