Renewable perfluorosulfonated ionomer carbon paste electrode for competitive homogeneous electrochemical immunoassays using a redox cationic labeled hapten.
A Nafion-loaded carbon paste electrode (CPE) was prepared and its square wave voltammetric response at a fresh surface was optimized using (ferrocenylmethyl)-trimethylammonium iodide (1) as a model of a cation that accumulates irreversibly. Low current responses could be obtained reproducibly without preconditioning at a CPE loaded with 1-octanol and neutralized Nafion. Use of the former made the method more sensitive, and use of the latter gave lower background currents and stable analytical response over a long period of Nafion-loaded CPE use. Under optimized conditions and using a 5 min accumulation period, the modified electrode gave a linear current response over the range of 5 nM to 1.5 microM 1, a detection limit of 5 nM 1 and a relative standard deviation of 5% for 0.5 microM 1. Using these optimized conditions, the competitive homogeneous immunoassay of phenytoin as a model hapten was carried out with cobaltocenium-labeled phenytoin as tracer. Clinical serum samples (5 microL) of phenytoin at therapeutic concentrations (20-80 microM) could be analyzed using this method.
Rapicault
S
S
Laboratoire d'Electrochimie Organique, Université Blaise Pascal de Clermont-Ferrand, Aubière, France.
Limoges
B
B
Degrand
C
C
D002138:Calibration; D004563:Electrochemistry; D006801:Humans; D007118:Immunoassay; D010672:Phenytoin / Q000097:blood*
D010672:Phenytoin