METHOD OF DETERMINING 2,6-DIMETHYLHYDROXYBENZENE IN BIOLOGICAL MATERIAL Russian patent published in 2024 - IPC G01N30/12 G01N30/14 G01N30/32 G01N30/34 G01N30/52 G01N30/72 G01N30/86 C07B63/02 

FIELD: biology; toxicological chemistry.

SUBSTANCE: disclosed is a method for determining 2,6-dimethylhydroxybenzene in a biological material, which consists in the fact that a biological object is ground, treated with a mixture of ethyl acetate and acetone, separate extracts are combined, the solvent from the combined extraction is evaporated, the residue is dissolved in a mixture of diethyl ether-hexane solvents, extracted twice with portions of aqueous-alkaline solution, extracts are combined, acidified, extracted twice with portions of ethyl acetate, extracts are combined, the combined extract is evaporated, chromatographed in a semi-preparative column silica gel L 40/100 mcm using a mobile phase, fractions of the eluate containing the analysed substance are combined, the solvent is evaporated, the residue is dissolved in an organic medium and determination is carried out by a physicochemical method, wherein the ratio of ethyl acetate and acetone in the mixture for treating the biological material is 8:2 by volume, before dissolving in the mixture of solvents diethyl ether-hexane in ratio of 1:1 by volume, the residue is repeatedly treated with methyl acetate, methyl acetate extracts are separated, combined, methyl acetate from combined extraction is evaporated, during chromatography in a semi-preparative column of silica gel L 40/100 mcm, a hexane-dioxane mobile phase is used in ratio of 9:1 by volume, after dissolving the residue in an organic medium, which is acetonitrile, the analyte contained in the acetonitrile is converted into a trimethylsilyl derivative, for which acetonitrile solution is treated for 30 minutes with N-methyl-N-(trimethylsilyl)trifluoroacetamide at temperature 70 °C, diluted with chloroform 25 times, and determination is carried out by a physicochemical method, which is chromatography-mass spectrometry, using a capillary column with length of 25 m and an inner diameter of 0.2 mm with a stationary phase of (5% phenyl)-methylpolysiloxane, using a carrier gas helium supplied with rate of 0.6 ml/min, and a mass-selective detector operating in electron impact mode, the initial temperature of the column thermostat is 70 °C, this temperature is maintained for 3 minutes, further temperature is programmed from 70 °C to 290 °C at rate of 20 °C per minute, the final temperature of the column is maintained for 10 minutes, the signal intensity is recorded, a mass spectrum is recorded by total ion current and amount of 2,6-dimethylhydroxybenzene is calculated based on the area of the chromatographic peak of its trimethylsilyl derivative.

EFFECT: invention provides higher detection sensitivity.

1 cl, 4 tbl, 3 ex