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Biomedical subjects

M R Smyth

Publications and source records attributed to M R Smyth.

At least 73 records · Page 4Linked to original sources

Determination of selenium in blood plasma and serum by flow injection hydride generation atomic absorption spectrometry.

A flow injection hydride generation atomic absorption spectrometric (AAS) method has been used to determine the selenium concentrations of human serum and plasma samples following digestion with nitric, sulphuric and perchloric acids. In the hydride generation process, reduction was carried out by sodium tetrahydroborate to produce a hydride that was atomized in a flame-heated atomisation cell. The method had a detection limit of 1.2 ng ml-1 and a sensitivity of 2.1 ng ml-1. Within-run precisions of 5.8% at 20 ng ml-1 and 4.5% at 80 ng ml-1, and between-run precisions of 4.8% at 69 ng ml-1 and 3.4% at 80 ng ml-1 were obtained. An inter-laboratory comparison study with a graphite furnace AAS method was carried out and the results showed excellent agreement. The flow injection method of sample introduction allowed the use of a sample volume of 330 microliters with an injection rate of 90 injections per hour.

Borohydrides↗

Purification of human glutamate dehydrogenase (GDH) and an adsorptive voltammetric investigation of the interaction of GDH with rabbit anti-human GDH antibody.

A procedure for the isolation of glutamate dehydrogenase (GDH) from human liver, which involves the use of ion-exchange chromatography on diethylaminoethyl cellulose and affinity chromatography on guanosine triphosphate conjugated to Sepharose 4B, is described. The adsorptive voltammetric behaviour of human GDH, bovine GDH and rabbit anti-human GDH antibody was optimised with respect to accumulation potential, accumulation time and scan rate. The lower limits of detection were 0.2 and 1.2 mg l-1 for human and bovine GDH, respectively, and the lower limit of detection for rabbit anti-GDH antibody was 0.04 mg l-1. The interaction of human GDH with rabbit anti-human GDH antibody was also examined using this method.

Animals↗

Adsorptive stripping voltammetric determination of pipemidic acid in human urine.

The electrochemical behaviour of pipemidic acid (8-ethyl-5,8-dihydro-5-oxo-2-(1-piperazinyl)-pyrido[2,3-d]pyrimidine-6- carboxylic acid), a well known antimicrobial agent used for urinary infections, was investigated by linear-sweep, differential-pulse and square-wave voltammetry at a hanging mercury drop electrode. Two reduction processes were observed in Britton-Robinson buffers at acid pH, whereas only one or two processes were observed in alkaline solutions, dependent on the pH of the buffer employed. Adsorptive effects were used to accumulate the drug on to the electrode. The adsorbed species were measured voltammetrically by using a cathodic process appearing at -0.76 V in 0.1 M HCIO4. Linear calibration graphs were obtained in the range 2.5 x 10(-9)-2.0 x 10(-7) M. A simple procedure of extraction was employed for the determination of the drug in urine samples.

Adsorption↗

Electrocatalytic detection of streptomycin and related antibiotics at ruthenium dioxide modified graphite--epoxy composite electrodes.

The application of ruthenium dioxide (RuO2) modified electrodes to the electrocatalytic detection of the saccharide-related antibiotics streptomycin, novobiocin and neomycin, at low fixed potentials, was investigated. The RuO2-modified graphite - epoxy composite electrodes give extremely stable and reproducible catalytic oxidation currents for these antibiotics at potentials as low as +0.2 V (versus Ag - AgCl). Rapid quantification at the micromolar level is therefore possible. Standard calibration graphs for streptomycin and neomycin yielded slopes of 4.43 and 0.08 nA microM-1 over the linear ranges of 1.5 x 10(-6) - 2.5 x 10(-4) and 1 x 10(-5) - 2 x 10(-3) M, respectively. Owing to its catalytic oxidation by the RuIII - RuIV couple, rather than the RuIV - RuVI transition (which catalyses the oxidation of streptomycin and neomycin), novobiocin could be detected at a lower (+0.2 V) potential, with a sensitivity of 1.31 nA microM-1. Detection limits of 1.5, 6.0 and 10 microM were obtained for streptomycin, novobiocin and neomycin, respectively. These catalytic surfaces can be renewed (by polishing), with a surface-to-surface reproducibility of 6.5% for the detection of 5 x 10(-5) M streptomycin. The analytical application of RuO2-modified carbon paste electrodes to the analysis of these antibiotics by flow injection was investigated, with a view to liquid chromatographic separation with electrochemical detection applications.

Catalysis↗

Retention characteristics in high-performance liquid chromatography of basic drugs and plasma extracts on an alumina column.

The retention characteristics of 35 drugs on an alumina column using buffered aqueous methanolic mobile phases have been investigated. It has been shown that the effect of pH on retention depends on the pKa of the drug and the type of buffer ion used; that there is an inverse relation between ionic strength and solute retention; and that a decrease in the amount of methanol in the mobile phase causes the drugs to be held longer on the column. The chromatographic system was also coupled to an on-line column-switching assembly to facilitate extraction of the drugs from plasma, and results are presented which indicate that the alumina column lends itself well to this convenient method of plasma clean-up.

Chromatography, High Pressure Liquid↗

Adsorptive stripping voltammetry of human chorionic gonadotropin and two of its specific antibodies.

The electrochemical behaviour of human chorionic gonadotropin (hCG), as well as rat anti-beta hCG and rabbit anti-beta hCG antibodies, has been studied using cyclic voltammetry and adsorptive stripping voltammetry. Conditions have been optimised for the determination of these species by differential pulse adsorptive stripping voltammetry with respect to accumulation potential, accumulation time, scan rate, pulse amplitude and drop size. This technique has also been used to investigate the interaction of hCG with each of its specific antibodies in solution.

Animals↗

A comparison of silica and alumina columns for high-performance liquid chromatographic separations of basic drugs and plasma constituents following on-line solid-phase extraction with column switching.

A comparison has been made of a silica and an alumina column with respect to retention behaviour in high-performance liquid chromatography of pure drug solutions and plasma profiles following on-line solid-phase extraction. Using methanol-ammonium nitrate buffer (80:20, v/v) as eluent, it was found that these systems lend themselves well to the column switching technique. Increasing the pH of the mobile phase produced similar shifts in retention for basic drugs on both columns, but fewer plasma interferents are retained on the alumina column as compared to the silica column under the same operating conditions.

Aluminum Oxide↗

Adsorptive stripping voltammetry of F(ab')2 and Fab fragments of anti-mouse immunoglobulin G.

The adsorptive stripping voltammetric behaviour of F(ab')2 and Fab fragments of anti-mouse immunoglobulin G is described. Conditions were optimized for the determination of these fragments with respect to accumulation potential, accumulation time, scan rate, pulse amplitude, drop size and electrolyte composition. The F(ab')2 and Fab fragments gave rise to behaviour similar to that reported previously for the intact immunoglobulin.

Adsorption↗

Silicone-grease-based immobilisation method for the preparation of enzyme electrodes.

An approach to the construction of amperometric biosensors based on the incorporation of an enzyme in silicone grease and using the grease to fill micropores on a graphite surface is described. The enzyme-grease electrode concept, illustrated with the enzyme tyrosinase, offers a very simple, rapid and inexpensive approach to the fabrication of enzyme electrodes. The tyrosinase electrode responds very rapidly to dynamic changes in the concentration of phenolic compounds. A response time (t95%) as low as 5 s has been determined. With flow injection, 120 samples per hour can be processed with a relative standard deviation of 2.4%. The electrode remains active for about 12 d. The detection limit for dopamine is 6 x 10(-6) M. This method of biosensor construction should be applicable to other enzyme-substrate systems.

Catechol Oxidase↗