METHOD OF DETERMINING 3-METHOXYHYDROXYBENZENE IN BIOLOGICAL MATERIAL Russian patent published in 2015 - IPC G01N33/50 G01N30/02 

FIELD: chemistry.

SUBSTANCE: biological material containing 3-methoxyhydroxybenzene is repeatedly (three times) treated for 45 minutes with an alkyl acetate in the form of methyl acetate; the separate extracts are combined; the solvent from the combined alkyl acetate extract is evaporated; the residue is repeatedly treated with acetone; the acetone extracts are combined, evaporated in an air current at 18-22°C, and then in an nitrogen current until complete removal of the solvent; the residue is dissolved in ether; the obtained solution is diluted with hexane in volume ratio of 1:1, extracted with a buffer solution with pH 12-13; the water-alkaline extract is separated, acidified to pH 2-3, saturated with sodium sulphate, extracted with diethyl ether; the ether extract is separated, dehydrated, evaporated in an air current at 18-22°C and then in a nitrogen current until complete removal of the solvent; the residue is dissolved in a hexane-dioxane-propanol-2 solvent mixture, taken in volume ratio of 20:5:1, subjected to chromatography in a silica gel macrocolumn L 40/100 mcm using a mobile phase of hexane-dioxane-propanol-2 in volume ratio of 20:5:1; eluate fractions containing the analyte are combined; the eluent is evaporated in an air current at 18-22°C and then in a nitrogen current until complete removal of the solvent; the residue is dissolved in dichloromethane, followed by determination using a chromatographic-mass-spectrometric technique using a capillary column with length of 25 m and internal diameter of 0.2 mm with a stationary phase with thickness (5% phenyl)-methylpolysiloxane, using a helium carrier gas, fed at a rate of 0.6 ml/min, and a mass-selective detector operating in electronic impact mode; the initial thermostat temperature of the column is 70°C; said temperature is maintained for 3 min, and the temperature is then raised from 70°C to 290°C at a rate of 20°C/min; the injector temperature is 250°C, the detector interface temperature is 300°C; the method also includes detecting the strength of the signal resulting from charged particles formed when bombarding the analyte coming from the capillary column and falling into an ion source which ionises an electron beam with energy of 70 eV; recording the mass spectrum on the full ion current and calculating the amount of 3methoxyhydroxybenzene from the area of the chromatographic peak obtained by detecting the signal from the characteristic molecular ion 124 m/Z.

EFFECT: high sensitivity.

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