Determination of baclofen in human plasma and urine by high-performance liquid chromatography with fluorescence detection

An isocratic reversed-phase column liquid chromatographic assay for baclofen was developed. Prior to analysis, derivatization of urine or deproteinized plasma with 4-chloro-7-nitrobenzofurazan (NBD-CI) reagent was employed. After solid-phase extraction on a small silica gel column, the mixture was chromatographed on a 10 microns Bondapak C18 column using methanol-water (45 + 55 v/v) as the mobile phase. The NBD-baclofen derivative was detected spectrofluorimetrically by monitoring the emission at 524 nm with excitation at 463 nm. The method was tested for suitability in routine analysis. After a single oral dose of 20 mg of baclofen, the plasma concentration and the cumulative urinary excretion of the drug were determined. The average recoveries were 99.5 and 98.5% for plasma and urine, respectively.