A FLUOROGENIC REAGENT, 3-(4,6-DIFLUOROTRIAZINYL)AMINO-7-METHOXYCOUMARIN, FOR THE DETERMINATION OF AMANTADINE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A sensitive method for determination of amantadine by high-performance liquid chromatography (HPLC) has been developed. Amantadine was derivatized quantitatively into a fluorescent compound through the amino group by treatment with 3-(4,6-difluorotriazinyl)amino-7-methoxycoumarin (FAMC) in benzene at 140 °C for 15 min. The derivative was subjected to HPLC on TSK gel ODS-120T (250 x 4.6 mm i.d.) with acetonitrile-water (5:1) as the mobile phase and monitored by fluorescence detection (Ex. 345 nm; Em. 410 nm). The limit of detection was 250 fmol in 100 μl of urine. This method was satisfactory with respect to recovery and precision to quantify amantadine spiked in urine. © 1990, The Pharmaceutical Society of Japan. All rights reserved.