Simple, rapid and sensitive determination of protionamide in human serum by high-performance liquid chromatography

A simple high-performance liquid chromatography method has been developed that allows the sensitive determination of protionamide (2-n-propyl-pyridine-4-carboxylic acid thioamide, PTH) in human serum. After pretreatment of the serum with trichloroacetic acid (TCA) and centrifugation the supernatants were neutralized using NaHCO3. PTH was separated on a Kromasil 100 C4 column (acetonitrile-sodium tetraborate buffer pH 8-dibutylamine) and determined photometrically at 291 nm. The lower limit of quantification for 300 microl serum precipitated with 60 microl TCA and injection of 50 microl was 27 microg/l and linearity was observed up to 15 mg/l.