HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION OFEBIRATIDE USING ETHYLAMINO-SULFONYL)-7-FLUORO-2,1,3-BENZOXADIAZOLE ASA FLUOROGENIC REAGENT

A sensitive high-performance liquid chromatography (HPLC) method with fluorescence detection has been developed and applied to the determina tion of ACTHJ(4-9) analogue (ebiratide), ethylamino-sulphonyl)-7-8uoro -2,1,3-benzoxadiazole (DBD-F) was used as a fluorogenic labelling reag ent. Ebiratide in 0.1 M berate buffer (pH 9.0) was reacted with DBD-F in acetonitrile solution at 50 degrees C for 30 min. After cooling, th e mixture was separated by a reversed-phase HPLC. Then DBD-ebiratide w as monitored at 580 nm with the excitation at 440 nm, The calibration curve was linear up to 10 pmol on column (r = 0.999) with the detectio n limit of 0.25 pmol (signal-to-noise = 3).