Biomedical subjects

M R Smyth

Publications and source records attributed to M R Smyth.

At least 19 recordsLinked to original sources

PubMed1992

Monitoring excitatory amino acid release in vivo by microdialysis with capillary electrophoresis-electrochemistry.

Capillary electrophoresis (CE) with electrochemical detection (ED) was used to determine extracellular levels of aspartate, glutamate and alanine in samples from the frontoparietal cortex of the rat which were obtained by microdialysis. The method was used to monitor the effect on the overflow of the excitatory amino acids aspartate and glutamate of an influx of high concentrations of potassium ion. Samples were derivatized with naphthalenedialdehyde-cyanide prior to analysis. Detection limits for aspartate and glutamate were 80 and 100 nM, respectively. CE-ED is extremely useful for the analysis of microdialysis samples because of the very small sample volumes required by this analytical technique. The use of ED provides the requisite sensitivity and allows verification of peak purity by voltammetry.

Amino Acids↗

PubMed1992

Analysis of terbutaline in human plasma by high-performance liquid chromatography with electrochemical detection using a micro-electrochemical flow cell.

A high-performance liquid chromatographic method is described for the determination of terbutaline in human plasma in the range 1-35 ng/ml. Detection was achieved using a carbon fibre micro-electrochemical detector and a column-switching system. The microelectrode cell has advantages over conventional glassy carbon electrode-based detection systems in that it is easy to prepare, flexible in its operation and suffers less trouble from problems such as air bubbles and leaks. Furthermore, it has a better detection limit for terbutaline (0.8 ng/ml) to that obtained using a conventional glassy carbon electrode flow detector (2 ng/ml). Sample clean-up was by on-line solid-phase extraction with column switching, providing a method which was sensitive and reproducible, where the mean overall coefficient of variation was 5.60% and drug recovery in excess of 86% at the concentration levels studied.

Calibration↗

PubMed1992

A study of intracolonic hydrogen and methane levels during colonoscopy.

Gas samples were obtained during colonoscopy for analysis. Patients were prepared with polyethylene glycol (PEG) (N = 23), phosphate enema (N = 34) and mannitol (N = 4). Air insufflation was used in all procedures. High concentrations of hydrogen were detected in 3 out of 38 gas samples in the PEG group, in 2 of 41 samples in the phosphate enema group and in one of the 8 samples in the mannitol group. All patients had a coexisting intracolonic oxygen concentration > 5%. The results suggest that potentially explosive concentrations of hydrogen may occur after conventional bowel preparations, and that insufflation of carbon dioxide during polypectomy should be a routine.

Colon↗

PubMed1992

Differential pulse voltammetric determination of sumatriptan succinate (1:1) in a tablet dosage form.

A voltammetric study of the oxidation of sumatriptan succinate (1:1) has been carried out at the glassy carbon electrode. This compound exhibited a single wave in Britton-Robinson buffer solutions of pH 2-11, with a maximum current at pH 5.0. The mechanism of oxidation was shown to be due to oxidation of the N-H group in the indole ring. Based on this study, a simple, rapid and sensitive voltammetric method was developed for the determination of the drug in a tablet dosage form.

Electrochemistry↗

PubMed1992

A lysine dehydrogenase-based electrode for biosensing of L-lysine.

An amperometric biosensor for L-lysine based on the recently isolated enzyme lysine dehydrogenase is described. Immobilization of the enzyme onto a platinum electrode is achieved via entrapment within a gelatin support on a cellulose membrane. Anodic detection (at 0.4 V vs. Ag/AgCl) is facilitated by the presence of a redox-mediating ferricyanide ion. The effect of experimental variables such as pH, enzyme loading, applied potential, cofactor and mediator concentrations were evaluated in order to optimize the analytical performance. A detection limit of 7 x 10(-8) M, and linearity up to 7 x 10(-4) M are reported. The fast response permits adaptation for flow injection operation with good precision (RSD = 1.9%) and high sample throughout (40 samples per hour). The high specificity offered by this new enzyme is indicated by the lack of interference by other L-amino acids, alcohols or carbohydrates.

Amino Acid Oxidoreductases↗

PubMed1991

High-performance liquid chromatographic determination of PEG 600 in human urine.

Polyethylene glycols (PEGs) are non-ionic, water-soluble synthetic polymers which have been widely used for many applications. Since they are of very low toxicity and are readily excreted in urine, PEGs in the molecular weight range 400-6000 have been used extensively in the study of intestinal physiology in man. A high-performance liquid chromatographic (HPLC) method has been developed for the determination of PEG 600 in human urine, which includes a pre-column derivatisation step. The dibenzoate derivatives of PEG 600 can be quantitatively prepared, and this, coupled with ultraviolet detection at 230 nm, has greatly improved the limit of detection for the determination of PEGs by HPLC. A suitable extraction procedure has also been developed which enabled PEG levels in urine to be monitored with much greater sensitivity than any previously reported method.

Benzoates↗

PubMed1991

High-performance liquid chromatographic determination of nifedipine, nicardipine and pindolol using a carbon fibre flow-through amperometric detector.

The electrochemical properties of the calcium-channel blockers, nifedipine and nicardipine, and the beta-blocking agent, pindolol, have been exploited for the determination of their concentrations in plasma samples. High-performance liquid chromatography (HPLC) separation was carried out on a cyanopropyl modified column and the drugs were detected in a flow-through carbon fibre microelectrode cell. The chromatographic system was coupled to a column-switching arrangement in order to perform on-line solid-phase extraction of the drugs from spiked human plasma. Preliminary investigations showed the response of the method to be linear over a range of 20-500 ng ml-1 in plasma with a limit of detection of approximately 15 ng ml-1 for each compound.

Carbon↗

PubMed1991

Liquid chromatography with amperometric detection of some sulphonamides and their N4-acetyl-metabolites in serum and urine.

Sulphonamides separated on a C18 LC column were detected at lower levels using amperometric detection at a glassy carbon electrode in comparison with UV detection at 258 nm. Whereas sulphonamides are detectable at a potential of +1.00 V, their N4-acetyl-metabolites required a potential of +1.25 V to be detected after their separation by LC. An interference commonly present in serum and urine, which co-eluted with one of the analytes, was detected at 1.25 V. This was overcome with an appropriate sample preparation in which 150 microliters of serum or 75 microliters of urine were first diluted to 1.5 ml with phosphate buffer (pH 3.0; 0.2 M). A 1.0 ml volume of this solution was then passed through an Extrelut 1 column. The analytes were eluted with dichloromethane, which was evaporated under vacuum, and redissolving the analytes in an appropriate volume of mobile phase, i.e. methanol-phosphate buffer (pH 6.7; 0.067 M) (25:75, v/v). For sulphamethoxazole (SMX) and sulfamethoxypyridazine (SMP) and their N4-acetyl-metabolites the calibration curves were linear between 1.5 x 10(-7) and 8 x 10(-6) M. The recovery ranged between 92.6 and 97.6% in serum and between 80.5 and 99.4% in urine. Detection limits were 10 times lower with amperometric detection than with UV detection. The method has been applied to the quantitation of SMX and SMP and their N4-acetyl-metabolites in serum and urine after their oral administration.

Acetylation↗

PubMed1991

Amperometric enzyme electrode for theophylline.

An amperometric biosensor for theophylline, based on the recently isolated enzyme theophylline oxidase, is described. The enzyme is entrapped, together with a ferricytochrome C cofactor, within a polymeric (Nafion) coating. The anodic detection (at +0.4 V versus Ag-AgCl) is facilitated by the addition of a redox-mediating hexacyanoferrate(III) ion. The influence of various experimental variables is described. The limit of detection is 2 x 10(-6) mol dm-3 theophylline, with linearity prevailing up to 3 x 10(-4) mol dm-3. The fast response and wash times permit rapid flow-injection measurements, with a frequency of 180 samples h-1 and a relative standard deviation of 3.0-4.0%. Prospects of using this electrode for clinical diagnostics are discussed.

Biosensing Techniques↗

PubMed1991

Reversed-phase high-performance liquid chromatographic determination of dexamethasone in bovine tissues.

A sensitive and selective high-performance liquid chromatographic procedure is described for the determination of the synthetic corticosteroid dexamethasone (DXM), in bovine muscle, kidney, liver and fat tissues, using methylprednisolone as the internal standard. Following extraction with ethyl acetate (muscle, kidney and liver) or diethyl ether (fat) and clean-up of the tissue extract, the drug residue was isolated using a C18 solid-phase extraction column. Separation of DXM was achieved by reversed-phase high-performance liquid chromatography with ultraviolet detection at 254 nm. By using this procedure, DXM levels as low as 0.01 mg kg-1 can be detected in muscle, kidney, liver and fat.

Adipose Tissue↗

PubMed1990

High-performance liquid chromatographic method for the comparison of tanning capacity of tannic acid batches used in the manufacture of pregnancy testing kits.

A high-performance liquid chromatographic (HPLC) method was developed for the quantitative comparison of various batches of tannic acid from the same manufacturer used to aid the binding of human chorionic gonadotropin to sheep erythrocytes in the manufacture of pregnancy testing kits. The tannic acids were separated by reversed-phase HPLC on a C18 column using gradient elution with aqueous methanolic eluents at low pH. A portion of the chromatogram corresponding to the compounds involved in the tanninisation process was integrated and a linear relationship was established between this peak area and tannic acid concentration. The correlation coefficient was greater than 0.993 even in the absence of an internal standard. Tanning capacity was evaluated on the basis of the amount of tannic acid which remained following incubation with a known quantity of erythrocytes. The application of this procedure to three batches of tannic acid is demonstrated.

Chromatography, High Pressure Liquid↗

PubMed1990

Evaluation of drug-free plasma profiles by high-performance liquid chromatography following on-line solid-phase extraction.

The effect of varying the type of column and eluent composition on drug-free plasma profiles was investigated. The study was based on a C18 and a CN column; methanol and acetonitrile were the organic modifiers used. The plasma profiles were evaluated quantitatively by measuring the number of interfering peaks greater than 8 . 10(-4) absorbance units in the area of interest along the chromatogram. Results were subjected to statistical treatment using a three-factor analysis of variance design. The three factors were the column, the type of organic modifier and either the percentage organic modifier, the pH or the ionic strength. Analysis of the data revealed that significant effects were seen with changing eluent composition, particularly with regard to the percentage of organic modifier, and that the observed effects were strongly dependent on the type of column and the type of organic modifier under consideration.

Acetonitriles↗

PubMed1990

Application of adsorptive voltammetry to assess the stability of modified alanine aminotransferases.

Alanine aminotransferase has been stabilized by using chemical modification with both bis(imidates) (of varying length) and succinic anhydride. The voltammetric behavior of the native enzyme and its various modified forms has been studied by using both cyclic voltammetry and differential pulse adsorptive voltammetry. A distinctive accumulation pattern was found for each of the stabilized enzymes at the static mercury drop electrode with respect to the native alanine aminotransferase. Adsorptive voltammetry was demonstrated to be a useful technique to assess the extent of chemical modification of this enzyme, which is indirectly related to their stability for use in biotechnological processes. The sue of differential pulse adsorptive voltammetry, after a preconcentration of the enzyme for 300 s at the electrode surface, has yielded a detection limit of 1.0 x 10(-9) M.

Alanine Transaminase↗

PubMed1990

High performance liquid chromatographic separation of cisplatin and its hydrolysis products on alumina and application to studies of their interaction with cysteine.

An alumina stationary phase has been assessed in the study of the retention behaviour of the anticancer drug cisplatin and its major hydrolysis products. Parameters such as buffer concentration in the mobile phase, pH, organic modifier and competing ion have been investigated in order to optimize chromatographic separation with ultraviolet detection. The separation scheme developed has been used to monitor the hydrolysis of cisplatin in aqueous and saline media, and to monitor the interaction of hydrolysed solutions of cisplatin with the amino acid cysteine. A new peak was observed in the chromatograms of such mixtures when they had been allowed to stand for periods of greater than 16 h and, from analysis of the data obtained, it was concluded that this new peak was due to a complex formed between the mono-aquo hydrolysis product of cisplatin and the amino acid.

Aluminum↗

PubMed1990

Adsorptive voltammetric investigation of the interaction of cisplatin with cystine and human serum albumin.

The interaction of the anti-cancer drug cisplatin with human serum albumin and cystine has been investigated using differential pulse adsorptive voltammetry. Based on an understanding of the voltammetric behaviour of these biological molecules, which rely on the presence of the disulphide groups within their molecular structure for their electroactivity, it has been postulated that binding of cisplatin to these molecules occurs at the disulphide bond. A fractional coefficient for the binding of cisplatin to human serum albumin at pH 7.4 was calculated to be 0.32. The reactivity of hydrolysis products of cisplatin was shown to be greater than that of the parent drug.

Adsorption↗

PubMed1990

Comparison of a calixarene-based ion-selective electrode with two automated analyzers for the clinical determination of sodium in blood plasma.

Neutral-carrier ion-selective electrodes based on methyl p-t-butylcalix[4]aryl acetate have been prepared that are responsive to sodium ions. The miniaturized catheter-type electrodes were obtained by dip-coating their porcelain tips in a PVC membrane cocktail. Examination of the general performance of the electrodes revealed excellent characteristics in terms of Nernstian response, selectivity, stability, reproducibility and response time. The results from the indirect potentiometric assessment of a large number of plasma samples with the electrodes showed a good correlation with the results from two automated analyzers (Technicon Smac 3, Hitachi 704) and with flame photometric data. Although inconsistencies were observed in the measurement of some plasma samples, the variance seemed to be method-dependent, and the overall performance of the electrodes showed promise as an alternative to the sodium glass electrode. Some factors influencing the standard potential of the measuring cell are discussed as a source of error.

Autoanalysis↗

PubMed1990

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↗

PubMed1990

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↗