Tuning a pyrazoline-based fluorogenic reagent, 3-naphthyl-1-(4-trifluoromethyl)-5-(4-carboxy phenyl)-2-pyrazoline for sensitive precolumn derivatization of aliphatic alcohols and trace analysis by RP-HPLC with fluorescence detection

A new member of the pyrazoline family, 3-naphthyl-1-(4-trifluoromethyl)-5-(4-carboxy phenyl)-2-pyrazoline has been evaluated as a precolumn derivatization reagent for the analysis of primary alcohols using HPLC. The simultaneous separation of eight alcohol derivatives (C₁–C₈) within 15 min was achieved on a reverse-phase C₈ column with an isocratic elution mode. The derivatives were detected with fluorescence at an emission wavelength of 470 nm when excited at 360 nm. The identification of the corresponding derivatives was carried out by LC–MS/MS and all showed their characteristic parent peak in negative ion mode. The proposed method was validated using normal analytical tools and was found to be excellent. As a preliminary application, our method was used to determine ethanol concentration in alcohol-containing chocolates and cough syrup.