DETERMINATION OF MITOMYCIN-C IN RABBIT OCULAR TISSUE AFTER TOPICAL ADMINISTRATION BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A high-performance liquid chromatographic method is described for the determination of mitomycin C (MMC) in rabbit ocular tissues. The optim ial conditions were 20 mM sodium phosphate buffer, pH 7.0 - methanol ( 70:30) in isocratic elution (flow rate 1 mL/min). The detection limit of MMC of this method is 0.02 mu g/g for conjunctive and sclera or 0.0 2 mu g/mL for aqueous humor. The effect of pH value and temperature on the stability of MMC was investigated. Acetonitrile was used for extr acting MMC from conjunctiva and sclera, and ethyl acetate for aqueous humor. Recoveries ranged from 49% to 92% over a wide range of concentr ations (0.2-400 mu g/g). The half-life of MMC, after cellulose sponge administration, is 0.58 hours for the conjunctiva and 0.45 hours for t he sclera. The peak concentration of MMC in aqueous humor is 1 hour.