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Abstract
The performance of amperometric and fluorescence detection vs UV detection in high performance liquid chromatography (HPLC) have been evaluated for monitoring indomethacin at trace levels in rabbit aqueous humour. Amperometric detection at a potential of +1.20V (vs Ag/AgCl) did not present any advantage over UV detection. However, fluorescence detection after liquid-liquid extraction with ethyl acetate and pre-column hydrolysis yielded limits of detection (2 μg l−1) and quantitation (5 μg l−1), lowered by a factor of 2 by comparison with UV detection. Validation results for a fluorescence method in aqueous humour are presented concerning specificity, linearity, accuracy, precision, analytical recovery, limits of detection and quantitation.