Determination of nodularin using immunoaffinity cartridge-high performance liquid chromatography

Abstract

An immuno-antigen of nodularin was prepared, and then a polyclonal antibody against nodularin was obtained from Japanese white rabbits immunized with the immuno-antigen. Immunoaffinity cartridges were prepared using CNBr-activated Sepharose 4B and the polyclonal antibody against nodularin and their coupling rate ranged from 77.5% to 90.2%. The cartridge load for nodularin and the recovery ranged from 70 ng to 98 ng and 71.8% to 97.2%, respectively. A method for trace level analysis of nodularin in water using the immunoaffinity cartridge and HPLC was established, its detection limit was 3 ng L⁻¹ and the linear range was from 10 to 500 ng L⁻¹.