A sensitive radioimmunoassay for budesonide in plasma.

Budesonide is a highly potent non-halogenated glucocorticoid with local anti-inflammatory properties. A sensitive radioimmunoassay for the measurement of the drug in unextracted plasma has been developed. Budesonide 21-hemisuccinate and budesonide 3-(O-carboxymethyl)oxime were conjugated to ovalbumin using the mixed anhydride method and antibodies to the haptens produced in sheep. The specificity of the antisera towards cortisol, and possible budesonide metabolites reflected the different sites of the attachment of the haptens to the carrier protein. An antiserum raised against the 3-(O-carboxy methyl)oxime conjugate was more specific (0.001% cross reaction) with respect to cortisol than the antiserum raised against the 21-hemisuccinate conjugate (0.344% cross reaction). Endogenous steroids at concentrations normally encountered in clinical samples, would not interfere with the measurement of budesonide. The radioimmunoassay was developed using the budesonide 3-(O-carboxy methyl)oxime antiserum, [3H]-budesonide and dextran coated charcoal phase separation. The theoretical limit of detection of the assay was 50 pg/ml. Budesonide was quantitatively recovered from normal human drug/free plasma at concentrations above 1 ng/ml (2.32 nmol/l) with a between batch variation of 12-15%. Budesonide was measured in the unextracted plasma of volunteers who had inhaled the drug from a pressurised aerosol spray.