An Enzyme-Linked Immunosorbent Assay for Determination of Dicyclanil in Animal Tissue

Abstract

Dicyclanil is a toxic, pyrimidine-derived insect growth regulator used in veterinary medicine for the prevention of myiasis. A competitive indirect enzyme-linked immunosorbent assay was developed firstly for the determination of dicyclanil in animal tissue. The antigen was prepared using the glutaraldehyde method and polyclonal antibodies were obtained by immunizing rabbits with a dicyclanil-bovine serum albumin conjugate. The antibody obtained was specific for dicyclanil with an IC₅₀ of 9 ng/mL. Recoveries from sheep tissue and liquid milk were in the range of 52.5–72.4%, with coefficients of variation between 4.7% and 11.9%. The results were further confirmed by high performance liquid chromatography–tandem mass spectrometry.

Keywords:

• Animal tissues
• Dicyclanil
• Enzyme-linked immunosorbent assay
• Polyclonal antibodies

Acknowledgments

This research was supported in part by the Project for Academic Human Resources Development in Institutions of Higher Learning under the Jurisdiction of the Beijing Municipality (PHR), and the National Eleventh Five-Year Scientific and Technological Support Plan (2011BAD34B01).

Notes

^a The MRL for fat has been recommended to be increased to 150 µg/kg (CVMP 2000).

^a Molar cross-reactivity (CR_molar) was calculated using units of nmol/mL for the IC₅₀ concentrations of the chemicals (Xu et al. 2009).

MR, Mean recovery; CV, coefficients of variation.

Xiaolin Hou and Shoujun Cao contributed equally to this work.