Biomedical subjects

T G Strein

Publications and source records attributed to T G Strein.

5 recordsLinked to original sources

PubMed2001

Discontinuous electrophoretic stacking system for cholate-based electrokinetic chromatographic separation of 8-hydroxy-2'-deoxyguanosine from unmodified deoxynucleosides.

The stacking and baseline-resolved separation of the oxidative damage marker, 8-hydroxy-2'-deoxyguanosine (8-OHdG), from unmodified deoxynucleosides in under 4 min is reported. Separations of 8-OHdG from 2'-deoxyadenosine, 2'-deoxycytosine, 2'-deoxyguanosine, and thymidine are accomplished using micellar electrokinetic capillary chromatography with sodium cholate. Importantly, the use of sulfate, intentionally added to the sample matrix, results in effective stacking of 8-OHdG and other analytes. This work extends electrokinetic stacking injection of neutral analytes to include deoxynucleosides. The procedure works well with either electrokinetic or hydrodynamic injection. The separation buffer and sample matrix composition were optimized to effect stacking conditions with an uncoated 50 microm fused-silica capillary. The lower limit of detection for the analytes is in the nanomolar range, and is more than an order of magnitude lower than without stacking. With 30 s (5.7 cm) electrokinetic injections, stacking and baseline separation of 8-hydroxy-2'-deoxyguanosine from the unmodified nucleosides is accomplished, even in the presence of a 400-fold excess of unmodified deoxynucleosides.

8-Hydroxy-2'-Deoxyguanosine↗

PubMed2001

Electrophoretically mediated microanalysis with small molecules: the Jaffé method for creatinine carried out in a capillary tube.

An eletrophoretically mediated microanalysis (EMMA) approach, used to perform online chemistry between two small molecules, has been characterized and optimized. The "plug-plug" type EMMA method involved electrophoretic mixing and subsequent reaction of nanoliter plugs of creatinine-containing samples and alkaline picrate (Jaffe reaction) within the confines of the capillary column, which acts as a microreactor. Analyses were performed by pressure injecting a plug of picrate followed by a plug of the creatinine-containing sample. A potential was then applied to electrophoretically mix the two reactants, and an incubation time of up to 6 min allowed the reaction to proceed prior to the application of a 27 kV separation potential with absorbance detection at 485 nm. The use of a 50 microm inner diameter(ID) extended light path capillary (150 microm pathlength) was found to be adequate for determining elevated levels of creatinine in human blood sera, but could not be used to quantify normal levels. Quantification of both normal and elevated levels of creatinine in sera was possible with a 75 microm ID high-sensitivity cell (1200 microm pathlength). Calibration plots using the latter for creatinine in human blood sera spanned the expected clinical range and were linear between 40 microM and 1.2 mM (r2 = 0.996) with an estimated limit of detection of 17 microM (signal-to-noise ratio S/N = 3). A quantitative comparison of results obtained with the reported EMMA method and accepted clinical methodology correlated very well (slope = 1.001).

Artifacts↗

PubMed2000

Determination of uremic toxins in biofluids: creatinine, creatine, uric acid and xanthines.

Rapid and accurate determination of small molecule metabolic end-products is vital for clinical diagnosis and study of many metabolic disorders and medical abnormalities. Chromatographic and electrophoretic techniques are attractive for clinical analyses because of the inherent ability to analyze multiple component biofluids and determine the analytes of interest with minimal interference from other species. This manuscript reviews recent (1990-present) developments in chromatography and electrophoresis methodology for the determination of creatinine, creatine, uric acid and xanthines in biofluids.

Body Fluids↗

PubMed1997

Determination of creatinine and other uremic toxins in human blood sera with micellar electrokinetic capillary electrophoresis.

We have been interested in the clinical use of capillary electrophoresis (CE) to monitor low-molecular-mass uremic toxins in body fluids. Creatinine, an important clinical marker for renal failure, is zwitterionic over a fairly wide pH range (pH 5-9) and can not be resolved from neutral components using free solution CE under these conditions. We report here a micellar electrokinetic capillary chromatography method using an sodium dodecyl sulfate-borate buffer system at pH 9.0 to determine creatinine levels in human serum. This method, performed on deproteinized sera, is also suitable for determining multiple ionic components. Moreover, this method compares favorably with an enzymatic method for creatinine performed in a clinical laboratory and thus appears to be a promising method in terms of potential clinical use.

Creatinine↗

PubMed1995

Capillary electrophoresis as a clinical tool. Determination of organic anions in normal and uremic serum using photodiode-array detection.

We report the use of free solution capillary electrophoresis to identify and quantify low-molecular-mass compounds found in normal and uremic serum as well as in hemodialysate fluid. The method reported provides a multicomponent analysis, allowing a single-step screening for more than 19 metabolites in less than 16 min. Serum samples from healthy individuals and from patients who have been diagnosed with chronic renal failure are analyzed using a borate buffer system at pH 9.0, and an extended light path capillary. Several ionic sample constituents are identified by electrophoretic mobility, UV spectra, and spiking with authentic standards. An analysis of the relative concentration of several metabolites, including hypoxanthine, pseudouridine, hippuric acid, and uric acid is presented. Each of these four metabolites is found in both normal and uremic serum samples (limits of detection 1 to 6 microM). Moreover, each of these metabolites is present at significantly elevated levels in uremic patients. The method reported is shown to have promising clinical utility for profiling serum sample constituents, and for quantitative determination of a few important metabolites.

Anions↗