Abstract
An improved GLC procedure was developed for the simultaneous determination of phenytoin and its metabolite, 5‐(p‐hydroxyphenyl)‐5‐phenylhydantoin, in plasma and urine following enzyme hydrolysis. After extraction, the drug, the metabolite, and the internal standard, 5‐(p‐methylphenyl)‐5‐phenylhydantoin, are measured by GLC with flame‐ionization detection as their respective methyl derivatives following flash‐heater methylation with tri‐methylanilinium hydroxide. The drug and metabolite give well‐resolved symmetrical peaks on a phenyl methyl silicone column, and the method has a sensitivity of 150 ng/ml of phenytoin and 125 ng/ml of the metabolite. GLC–mass spectral evidence is presented for the formation and intact determination of methyl derivatives of the drug, its metabolite, and the internal standard.
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