Dual-label time-resolved fluoroimmunoassay of psychopharmaceuticals and stimulants in serum

Abstract

A new method to measure two different drugs simultaneously by time-resolved fluoroimmunoassay (TR-FIA) has been developed. In the TR-FIA reported here, psychopharmaceuticals [chlorpromazine (CPZ) and desipramine (DSP)] and methamphetamine (MA) contained in serum are assayed by a combined use of a new europium (Eu) chelate and a samarium (Sm) chelate, as labels. The drug concentrations were determined by the competition between a labeled antigen with Eu 3+ or biotin and a sample antigen. A microtiter plate coated with a mixture of rabbit IgGs (anti-MA and anti-CPZ or anti-MA and anti-DSP) was used. In the assay of MA and CPZ, Eu 3+ labeled MA–bovine serum albumin conjugate (MA–BSA) and biotinylated CPZ–BSA were added to the well with their non-labeled standard solutions or samples. MA was assayed by measuring the fluorescence intensity of Eu 3+ at 615 nm. After incubation of the Sm 3+ labeled streptavidin, CPZ was assayed by measuring the fluorescence of Sm 3+ at 643 nm. In the assay of MA and DSP, Eu 3+ labeled DSP–BSA and biotinylated MA–BSA were used. In our dual-assay, the minimum detection limits of these drugs were 1ng/ml for MA, 10 ng/ml for CZP and 10 ng/ml for DSP. Since the simultaneous detection of different drugs by TR-FIA is time and sample saving, the method can be employed in rapid and sensitive screening tests.