METHOD OF DETECTING 2-METHOXY-4-ALLYLHYROXYBENZENE IN BIOLOGICAL MATERIAL Russian patent published in 2012 - IPC G01N33/48 G01N30/02 

FIELD: chemistry.

SUBSTANCE: biological object is treated twice for 30 minutes with ethyl acetate; ethyl acetate extracts are combined; ethyl acetate is evaporated; the residue is dissolved in hexane and extracted with a buffer solution with pH 12-13; aqueous-alkaline extracts are separated and acidified to pH 2-3, saturated with sodium sulphate, extracted with diethyl ether; the ether extract is separated, dehydrated, evaporated at 18-22°C in an air current and then in a nitrogen current until complete removal of the solvent; the residue is dissolved in a mixture of solvents, chromatographed in a macrocolumn with silica gel L 40/100µ using a mobile phase; the eluate fractions containing the analysed substance are combined; the eluent is evaporated at 18-22°C in an air current and then in a nitrogen current until complete removal of the solvent; the residue is dissolved in hexane and determination is carried out using a chromato-mass-spectrometric method using a capillary column with length of 30 m and inner diameter of 0.25 mm, which contains a stationary phase, using a helium carrier-gas fed at a rate of 1 ml per minute, and a mass-selective detector operating in electron impact mode; temperature of the injector is 280°C, temperature of the interface is 260°C, temperature of the quadropole is - 150°C, temperature of the ion source is 230°C; intensity of the signal caused by charged particles formed during bombardment with a 70 eV electron beam of the analysed substance, which comes out the capillary column and falls into the ion source, is recorded; the mass spectrum on the full ion current is recorded and the amount of 2-methoxy-4-allylhydroxybenzene is calculated from the area of the chromatographic peak; wherein after evaporation of ethyl acetate, the residue is repeatedly treated with acetone, acetone extracts are combined, evaporated first in an air current at temperature 18-22°C and then in a nitrogen current until complete removal of the solvent; before chromatography in the macrocolumn, the residue is dissolved in a mixture of hexane-diethyl ether solvents, taken in volume ratio 6:4; the stationary phase for chromatography in the macrocolumn is a mixture of hexane-diethyl ether solvents in volume ratio 6:4; during chromato-mass-spectrometric determination, the stationary phase of the capillary column is (5%-phenyl)-methylpolysiloxane, the initial temperature of the column is 50°C, temperature is programmed from 50°C to 200°C at a rate of 10°C per minute.

EFFECT: higher sensifility of the analysis.

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