부산대학교 도서관

A fluorometric method has been developed for quantitation of α-[(t-butylamino)methyl]-3-methoxy-4-hydroxybenzylalcohol (I). The method is based on the fluorescence of dansyl-derivative of I after separation on a thin-layer chromatographic plate, and can be applied to the determination of I in the concentration range of (0.040.4) ppm in rat plasma. The procedure is as follows. To 0.5 ml of rat plasma was added 4 ml of ethanol, and the mixture was centrifuged. To 3.5 ml of supernatant was added 1 ml of 0.01 M sodium carbonate and 0.5 ml of 0.5 % dansylchloride acetone solution, and the mixture was reacted for 30 min at room temperature (2225)°C. Then the reaction mixture was condensed to (0.050.1) ml under reduced pressure at 40°C. To the residue was added 2.5 ml of water and 2 ml of ether, and the mixture was shaken for 3 min. After centrifugation, ether extract was poured upon a mini-column packed with silica gel [Merck, (0.050.2) mm, 100 mg], and the column was washed with 8 ml of ether and eluted with 5 ml of acetone. The acetone eluent was evaporated to dryness under reduced pressure at room temperature. The residue was dissolved in 50 μl of acetone and its 10 μl was spotted on the silica gel plate. The plate was developed with a mixed solvent of benzene-triethylamine-acetone (4 : 1 : 1, v/v) for 30 min. After drying the plate for 10 min in air, the fluoresence intensity of dansyl-derivative of I (Rf 0.17) was measured at 520 nm with excitation at 350 nm.