title: Spectrophotometric determination of carp vitellogenin using a sequential injection analysis technique equipped with a jet ring cell.
abstract: A sequential injection analysis (SIA) technique, in which antibody-immobilized microbeads were transferred to a jet ring (JR) cell, was used in determination of carp vitellogenin (Vg). The determination is based on a sandwich immunoassay in which two types of reactions between anti carp Vg antibodies and carp Vg are used. Namely, the antibody for the first reaction step was immobilized on microbeads (Sephadex beads), and an antibody labeled with a horseradish peroxidase (HRP) was used in the second step of the reaction. A mixed solution of hydrogen peroxide and o-phenylenediamine (OPD) was used as the source of the chromophore in the reaction. The microbeads-immobilized antibody, Vg analyte, HRP-labeled anitbody and the color developing solution were introduced automatically into the JR cell of the SIA system in a programmed sequence, and the absorbance of the oxidized OPD product was used to determine the amount of Vg present. The optimal incubation times for the immuno-raction for the first and the second steps were determined at 120 and 60 min, respectively, taking into account the sensitivity to the Vg determination. Under these conditions, a good linear correlation was obtained between Vg concentration and the absorbance of the oxidized OPD. The lower detection limit for the determination of Vg was about 5 ng ml(-1) in this system. The method developed here represents a simple, accurate method for the determination method of Vg.
authors.affiliation: Department of Chemical Systems and Engineering, Graduate School of Engineering, Kyushu University, Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Soh
authors.forename: Hideshi
authors.initials: H
authors.lastname: Nishiyama
authors.forename: Keiko
authors.initials: K
authors.lastname: Mishima
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Imato
authors.forename: Takashi
authors.initials: T
authors.lastname: Masadome
authors.forename: Yasukazu
authors.initials: Y
authors.lastname: Asano
authors.forename: Youichi
authors.initials: Y
authors.lastname: Kurokawa
authors.forename: Hisao
authors.initials: H
authors.lastname: Tabei
authors.forename: Saeko
authors.initials: S
authors.lastname: Okutani