Simple spectrofluorometric determination of p-aminobenzoic and p- aminosalicylic acids in biological fluids by use of terbium-sensitized luminescence

Abstract

A novel, sensitive, and selective method has been developed for determination of p-aminobenzoic (PABA) and p-aminosalicylic (PAS) acids in the N-benzoyl-L-tyrosyl-PABA/PAS test, PAS is measured as a ternary complex with terbium and EDTA (λ(ex) = 324 nm, λ(cm) = 546 nm) in alkaline aqueous solution (pH~ 12.6), whereas both compounds (PABA and PAS) are measured as ternary complexes with terbium and tri-n-octylphosphine oxide (λ(cs) = 292 nm, λ(cm) = 546 nm) in weakly acidic aqueous solution (pH~ 5.5). We investigated and implemented optimum conditions for formation of these complexes, yielding respective detection limits for PABA and PAS of 0.07 and 0.02 μmol/L and ranges of application of 0-10 and 0-40 μmol/L (final concentration). The method has been successfully applied to determinations of PABA and PAS in urine and, after alkaline hydrolysis, to determinations of PABA in serum that has been deproteinized with acetonitrile. Within-run imprecision of the PABA determination ranges from 0.8% to 4.2% for urine samples and from 3.9% to 8.2% for serum samples; day-to-day imprecision varies from 3.2% to 10% for serum samples.