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Biomedical subjects

G G Guilbault

Publications and source records attributed to G G Guilbault.

At least 73 records · Page 4Linked to original sources

Determination of plasma glucose with use of a stirrer containing immobilized glucose dehydrogenase.

A stirrer containing immobilized glucose dehydrogenase has been successfully used for determining glucose in plasma. The device is usable for at least two months and for about 500 assays. The reaction was measured kinetically and linearity was observed to 4 g of glucose per liter. Tested with aqueous glucose and with deproteinized plasma, within-day and day-to-day precision were good. Interference and method-comparison (hexokinase method) were examined. The performance of this system makes the technique useful and attractive for routine use in small-volume clinical laboratories.

Blood Glucose↗

Measurement of glucose in plasma, with use of immobilized glucose oxidase and peroxidase.

We describe a novel fluorometric technique for simple, rapid, and economical assay of glucose by use of immobilized glucose oxidase and peroxidase. A cylindrical magnetic stirrer was specially designed to hold the immobilized enzymes firmly and to allow the reaction minzymic transformation quickly. Blood plasma can be assayed directly with no pretreatment. Ascrobic acid, uric acid, and creatinine in concenctrations of 0.25, 0.25, and 0.2 g/liter, respectively, did not interfere. The linearity of the assay was extended to 4.0 g of glucose concentrations. Results by our technique and by the otoluidine or hexokinase methods agreed well. The immobilized enzymes are stable for several months and can be used for several hundred highly accurate and reproducible assays.

Aspergillus niger↗

Enzymatic, fluorometric assay of alpha-amylase in serum.

An enzymatic, fluorometric method for kinetic assay of serum alpha-amylase (1,4alpha-D-glucan 4-glucanohydrolase, EC 3.2.1.1) is described. A soluble starch is used as substrate, in tris(hydroxymethyl)methylamine buffer. All measurements are made in Pyrex cuvettes at 37 degrees C, with a reaction volume of 1.16 ml. The assay is based on the following reaction sequence: (see article) The rate of appearance of fluorescence of NADH (lambdaex = 365 nm, lambdaem = 460 nm), developed in the indicator reaction (4), is measured and equated to the activity of alpha-amylase in serum. A calibration plot of the change of fluorescence per min vs. enzyme concentration shows a good proportionality in the range of 0.50-5.0 kU/liter.

Amylases↗

An alternative method for the determination of uric acid in serum.

A novel fluorometric method for the determination of uric acid based on the coupled reactions of uric acid with uricase and peroxidase to form highly fluorescent 2,2'dihydroxy-3,3'-dimethoxy-biphenyl-5,5'-diacetic acid is described. The calibration curve was constructed from a series of standard uric acid solutions vs. the corresponding relative fluorescence. It was linear up to 15 mg/dl (0.9 mmol/l). The serum or plasma samples must be deproteinated with (absolute) ethanol before assay and its uric acid content can be obtained from the calibration curve. This method is rather simple, having good precision and accuracy. High level ascorbic acid in the sample falsely elevates uric acid concentration. Comparison of the results obtained on the patient sera with a colorimetric phosphotungstate method and a standard enzymatic spectrophotometric method gave coefficients of correlation of 0.908 and 0.986, respectively.

Ascorbic Acid↗