METHOD OF DETERMINING 2,6-BIS[BIS(BETA-HYDROXYETHYL)AMINO]-4,8-DI-N-PYPERIDINO-PYRIMIDO(5,4-D)PYRIMIDINE IN BIOLOGICAL MATERIAL Russian patent published in 2008 - IPC G01N33/49 G01N33/48 

FIELD: analytical methods in toxicological chemistry.

SUBSTANCE: invention relates to methods for determining title compound in biological material for use in practice of chemicotoxicological and clinical laboratories. Biological tissue is disintegrated, two times infused with acetone each time for 30 min, resulting extracts are combined, combined extracts are filtered, filtrate is evaporated to give dry residue, the latter is dissolved in chloroform and extracted with 0.1 hydrochloric acid, acid extract is washed with diethyl ether, alkalinized with 10% sodium hydroxide solution to pH 8-10, saturated with sodium chloride, and extracted with ethylacetate. Ethylacetate extract is separated, dehydrated, and evaporated to give residue, which is dissolved in mixture acetonitrile/1 N sulfuric acid (8:2 v/v), and chromatographed on column packed with sorbent "Silasorb C-18" having particle size 30 μm while using acetonitrile/1 N sulfuric acid (8:2 v/v) as mobile phase. Fractions of eluate containing subject substance are combined and optical density of combined eluate is measured at wavelength 286 nm.

EFFECT: reduced analysis time, increased degree of recovery, accuracy, and sensitivity of determination.

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