METHOD FOR DETERMINING ADRENALINE, NORADRENALINE, DOPHAMINE AND DIHYDROXIPHENYLALANINE CONCENTRATION IN SINGLE PORTION OF BIOLOGICAL MATERIAL Russian patent published in 2001 - IPC

FIELD: medicine. SUBSTANCE: method involves separating adrenaline, noradrenaline, dophamine and dihydroxyphenylalanine from biological material by alumina-mediated adsorption, elution, oxidation, with following concentration of these substances being determined from their fluorescence intensities. Biological material quantity is selected to be equal to 0.4 ml. The same volume of 5% sodium ethylenediaminetetraacetate solution is added to its acidic extract. Solution pH is brought to 8.2-8.5 using 0.1% ammonia solution. The resulting solution is introduced into chromatography column with alumina. Its height-to-diameter ratio is selected to be equal to 1. Elution with 1 ml of 0.25 N acetic acid solution is carried out. This solution volume is divided into 0.5 ml test sample and 0.5 ml of control one. 0.15 ml of sodium ethylenediaminetetraacetate and 0.12 ml of 0.33 M trisubstituted sodium phosphate to 6.0-6.5 pH. Adrenaline, noradrenaline, dophamine and dihydroxiphenylalanine oxidation is carried out with 0.1 ml of 0.1 N iodine solution during 10 min. Then the solution is neutralized with 0.1 ml of alkaline sulfite and 0.05 ml of acetic acid is added next to it. Then the solution is placed into water bath for 5 min and cooled. Fluorescence of control and test samples is measured at excitation wavelength λ_В being equal to 313 nm, 313 nm, 365 nm, 436 nm and fluorescence wavelength λ_Ф equal to 365 nm, 405 nm, 510 nm, 510 nm, respectively. Fluorescence of the standard solution control sample of the substances under study are compared to the fluorescence intensities of the standard solution test sample of the same substances with the mentioned excitation and fluorescence wavelength values set. EFFECT: higher sensitivity; less laboriousness. 5 cl