Simplified fluorometric assay for diphenylhydantoin in plasma.

The fluorometric benzophenone procedure for diphenylhydantoin has been simplified by eliminating the need for preliminary extraction of plasma with an organic solvent. Assays are done directly on 0.2 ml of plasma by treating it with alkaline permanganate to form benzophenone, extracting the benzophenone with heptane, and then shaking the heptane layer with sulfuric acid and measuring the fluorescence of the acid layer. The assay is highly reproducible, and adequately sensitive to detect 1 mug of the drug per milliliter of plasma. Fluorometric and gas-liquid chromatographic assays of 154 plasmas gave results that were not significantly different, even in the presence of phenobarbital, other barbiturates and anticonvulsant drugs, or of various tranquilizers and other commonly used drugs. The assay is rapid, the unit cost per assay low.