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Sensitive, fast, and specific immunoassays for methyltestosterone detection.

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Area of Science:

  • Veterinary Science
  • Analytical Chemistry
  • Food Safety

Background:

  • Methyltestosterone (MT) is an anabolic steroid sometimes misused in animal husbandry.
  • Accurate detection of MT residues in animal feed is crucial for regulatory compliance and consumer safety.
  • Existing detection methods can be time-consuming or require specialized equipment.

Purpose of the Study:

  • To develop and validate sensitive immunoassays for detecting methyltestosterone (MT) residues in animal feed.
  • To establish rapid and reliable tools for routine monitoring of MT contamination.
  • To ensure the safety and quality of animal feed products.

Main Methods:

  • Development of an indirect competitive enzyme-linked immunosorbent assay (icELISA) utilizing a specific monoclonal antibody.
  • Creation of an immunochromatographic strip assay for rapid, on-site MT detection.
  • Validation of assay performance using parameters such as limit of detection, specificity, and recovery rates.
  • Comparison of immunoassay results with gas chromatography-mass spectrometry (GC-MS) for accuracy.

Main Results:

  • The optimized icELISA demonstrated high specificity for MT with no cross-reactivity to eight analogues.
  • The icELISA achieved a limit of detection of 0.045 ng/mL and a half-inhibition concentration of 0.26 ng/mL.
  • Recovery rates for MT in animal feed using icELISA ranged from 82.4% to 100.6%, aligning with GC-MS results.
  • The novel immunochromatographic strip assay showed a visual cut-off value of 2.5 ng/g in fish and pig feed.

Conclusions:

  • The developed icELISA and immunochromatographic strip assay are effective and highly specific tools for detecting MT residues in animal feed.
  • These immunoassays offer rapid, sensitive, and reliable alternatives for routine monitoring and regulatory purposes.
  • The findings contribute to enhanced food safety by enabling efficient detection of prohibited substances in animal feed.