Development and Validation of an Indirect Competitive ELISA for Cyenopyrafen Detection Using a Novel Monoclonal Antibody

Cyenopyrafen is an effective pyrazole acaricide used for mite control on various crops. This study generated high-affinity monoclonal antibodies specific to cyenopyrafen and developed a high-throughput indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) validated for sensitivity, specificity, and reproducibility. The ic-ELISA demonstrated an IC 50 of 32.75 ng/mL and a dynamic range of 2.33-293.19 ng/mL, with negligible cross-reactivity (<0.1%) with related acaricides. Recovery rates in citrus samples were 88.18%–105.89%. A strong correlation (R 2 > 0.95) was detected between the results of ic-ELISA and UPLC-MS/MS, highlighting the assay’s reliability. This immunoassay offers a reliable approach for the determination of cyenopyrafen in the tested citrus and apple samples, and may serve as a basis for broader applications with potential utility in regulatory frameworks.