Sensitive Time-Resolved Fluorescence Immunoassay for Quantitative Determination of Oxyfluorfen in Food and Environmental Samples

The direct and indirect competition time-resolved fluorescence immunoassays (dc-TRFIA, ic-TRFIA) were established by combining the autofluorescence properties of lanthanide europium (Eu) with the monoclonal antibody of oxyfluorfen. The purified Eu antibody was optimized and the conditions such as the working concentration of the Eu antibody, monoclonal antibody, and working buffer were optimized. In the optimal condition, the IC(50) of dc-TRFIA was 10.27 ng/mL, the lowest detection limit IC(10) was 0.071 ng/mL, the detection range (IC(10)-IC(90)) was 0.071–1074.3 ng/mL, and the detection range (IC(10)-IC(90)) and IC(50) of ic-TRFIA were 0.024–504.6 and 2.76 ng/mL, respectively. The comparison showed that the sensitivity and detection limit of ic-TRFIA were superior to dc-TRFIA. The cross reaction (CR) tests showed that the CR with other oxyfluorfen structure analogs was <0.02%, except that there was a certain CR with the benzofluorfen (CR = 11.58) and the bifenox (CR = 8.23%). The average recoveries of ic-TRFIA were 74.6–108.3%, and the RSDs were between 2.1 and 10.9%, in the addition recovery test with five substrates. The results of the correlation test with the real samples of GC-ECD showed that they were highly correlated (y = 0.975x – 0.4446, R(2) = 0.9901), which proved that the TRFIA method established in this study had high reliability and accuracy and could be used in environment and agricultural products for rapid detection of oxyfluorfen residues.