Determination of oxolinic acid in the bryophyte Fontinalis antipyretica by liquid chromatography with fluorescence detection

A large amount of oxolinic acid administered in freshwater fish farms reaches the environment. In order to allow environmental monitoring, an HPLC method to determine oxolinic acid in the bryophyte Fontinalis antipyretica was developed. Nalidixic acid was used as an internal standard. Oxolinic and nalidixic acids were extracted by cracking the bryophytes in liquid nitrogen with 0.1 M acid oxalic solution in ethyl acetate and a liquid-liquid clean up procedure was then performed. Mobile phase was a 0.02 M orthophosphoric acid aqueous solution-acetonitrile mixture (70:30, v/v). The stationary phase was 5 microm PuroSpher RP18e and quinolones were detected by fluorescence. The linearity, accuracy and precision of this method were demonstrated by a validation assay. The limits of detection and quantitation were 1 and 10 ng/g respectively. The linearity range was 10 to 500 ng/g. This method was applied to a 25 days experimental study performed with the bryophyte Fontinalis antipyretica.