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High-performance liquid chromatographic assay for antiinflammatory agents diclofenac and flurbiprofen in ocular
1Department of Ophthalmology, University of Colorado Health Sciences Center, Denver 80262.
Abstract:
An HPLC assay for diclofenac (DC) and flurbiprofen (FP) in a 100-microliters sample of aqueous humour is presented. After acetonitrile extraction, the residue is analyzed using a reversed-phase octyl column and ultraviolet detection. The method is simple and reproducible. Excellent selectivity and resolution are achieved using an acetic acid-acetonitrile-triethylamine mobile phase. The lower limit of detection, defined as the amount of drug required to produce a peak twice the threshold, was ca. 0.3 ng DC on column and ca. 0.4 ng on column FP. The utility of the method is demonstrated by determining drug levels in aqueous humour of normal rabbits and of patients undergoing cataract surgery. The assay should be applicable to other antiinflammatory agents.