Determination of fluoranthene by antigen-coated indirect competitive real-time immuno-PCR assay

Abstract

A reliable and sensitive antigen-coated indirect competitive real-time immuno-PCR (RT-IPCR) assay was developed for the determination of fluoranthene (FL). 9-Fluoranthenebutanoic acid, γ-oxo-(FLA) was synthesized as the hapten for FL. Mixed anhydride reaction (MAR) was used to couple the FLA to ovalbumin (OVA) to form artificial coating antigen. Active ester method (AEM) was used to couple the FLA to bovine serum albumin (BSA) to form artificial immune antigen. Male New Zealand white rabbits were immunized with immune antigen to obtain polyclonal antibodies (pAbs), with which, a novel RT-IPCR assay for determination of FL was described. Under best conditions, FL can be determined in a concentration range between 10 fg mL⁻¹ and 100 pg mL⁻¹ with a detection limit of 5 fg mL⁻¹. The cross-reactivities of the anti-FL antibody to seven structurally related compounds were below 11%. Some environmental water samples were analyzed with satisfactory results, which shows a good accuracy and suitability to analyze FL in environmental water. Recovery was lower or higher with agitation but would still be acceptable for use in an on-site field test to provide rapid, semi-quantitative, and reliable test results for making environmental decisions.