Sensitive enzyme immunoassay for screening methandienone in dietary supplements

ABSTRACT

Methandienone is a synthetic exogenous steroid which, like other anabolic steroids, is strictly regulated in many countries. In recent years, increasing numbers have been detected of illegal additions into dietary supplements of methandienone and other anabolic androgenic steroids (AAS). In this work, a competitive indirect enzyme-linked immunosorbent assay (ELISA) has been constructed for the detection of methandienone using an antiserum against methandienone. Under optimal experimental conditions, the ELISA achieved a limit of detection of 0.04 ± 0.01 µg.g⁻¹. The obtained intra- and inter-day coefficients of variation were less than 8%. The developed ELISA was applied in the analysis of real dietary supplement samples. To minimise the effect of the sample matrix, the sample extracts were simply diluted before addition into the immunoassay. The achieved recovery values were around 100%. Results obtained from the ELISA correlated well, both in terms of accuracy and precision, with those obtained by UHPLC-MS/MS (reference method). The presented ELISA could be successfully applied for the simple screening of dietary supplements.

Graphical Abstract

KEYWORDS:

• Methandienone
• anabolic steroid
• enzyme-linked immunosorbent assay (ELISA)
• screening method
• dietary supplements

Disclosure statement

No potential conflict of interest was reported by the authors.

Ethical Approval

All applicable international, national and/or institutional guidelines for the care and use of laboratory animals were followed.

Informed Consent

Informed consent was obtained from all individual participants included in the study.