PMID: 6110775Apr 1, 1981Paper

Determination of fluphenazine, related phenothiazine drugs and metabolites by combined high-performance liquid chromatography and radioimmunoassay

The Journal of Pharmacology and Experimental Therapeutics
S A Goldstein, H Van Vunakis

Abstract

Antibodies have been produced in rabbits immunized with a fluphenazine succinate-human serum albumin conjugate. By radioimmunoassay it is possible to quantify fluphenazine (FPZ), related phenothiazine drugs and several of their metabolites at the femtomole level. As little as 370 fmol (160 pg) of FPZ can be detected and up to 0.4 ml of plasma can be added to the incubation mixture (final volume = 1.1 ml). The phenothiazine heterocyclic nucleus is immunodominant and determines the specificity of the antiserum. When a parent drug cross-reacts significantly with antibody, its 7-hydroxide, N-oxide and N-10 side chain altered metabolites can also be determined by the assay. The 8-hydroxide, sulfoxide and 7-hydroxyglucuronide metabolites are not detectable unless present in large amounts. High-performance liquid chromatography was used to separate phenothiazine drugs and metabolites. Since the antiserum has broad specificity, a combined high-performance liquid chromatography and radioimmunoassay procedure permits the identification and quantification of a phenothiazine drug and its serologically reactive metabolites. Patterns of high-performance liquid chromatographic elution and extent of immunologic cross-reaction are characteristi...Continue Reading

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