A radioimmunoassay for the determination of tipredane in human plasma

A sensitive method for the determination of tipredane in human plasma has been developed. Prior to radioimmunoassay, proteins are removed from plasma by precipitation with acetonitrile. A [125I]iodohistamide derivative of tipredane is used as a heterogeneous radioligand. The primary antiserum is raised in sheep against a tipredane (O-carboxymethyl) oxime-bovine serum albumin conjugate. Donkey anti-sheep IgG antiserum is used as the secondary antiserum in a double antibody separation procedure. The limit of quantification of the method is 1 ng ml-1 for 500 microliters of plasma. There is no significant interference from either established or putative metabolites of tipredane, endogenous steroids or a wide range of other drugs. The assay was used to determine the concentrations of tipredane in plasma samples collected in pilot clinical studies.