Dual-label time-resolved fluoroimmunoassay of psychopharmaceuticals and stimulants in serum.
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.
Kimura
H
H
Department of Forensic Medicine, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo, Japan.
Mukaida
M
M
Wang
G
G
Yuan
J
J
Matsumoto
K
K
D000818:Animals; D000929:Antidepressive Agents, Tricyclic / Q000097:blood*; D014150:Antipsychotic Agents / Q000097:blood*; D002138:Calibration; D000697:Central Nervous System Stimulants / Q000097:blood*; D002746:Chlorpromazine / Q000097:blood*; D003891:Desipramine / Q000097:blood*; D015200:Fluoroimmunoassay / Q000295:instrumentation / Q000379:methods*; D008694:Methamphetamine / Q000097:blood*; D011817:Rabbits; D012680:Sensitivity and Specificity; D015813:Substance Abuse Detection / Q000379:methods*; D014116:Toxicology / Q000379:methods*
D000929:Antidepressive Agents, Tricyclic; D014150:Antipsychotic Agents; D000697:Central Nervous System Stimulants; D008694:Methamphetamine; D003891:Desipramine; D002746:Chlorpromazine