Novel dummy molecularly imprinted polymer for simultaneous solid-phase extraction of stanozolol metabolites from urine

Anal Bioanal Chem. 2024 Jun;416(14):3335-3347. doi: 10.1007/s00216-024-05285-x. Epub 2024 Apr 25.

Abstract

Stanozolol, a synthetic derivative of testosterone, is one of the common doping drugs among athletes and bodybuilders. It is metabolized to a large extent and metabolites are detected in urine for a longer duration than the parent compound. In this study, a novel dummy molecularly imprinted polymer (DMIP) is developed as a sorbent for solid-phase extraction of stanozolol metabolites from spiked human urine samples. The optimized DMIP is composed of stanozolol as the dummy template, methacrylic acid as the functional monomer, and ethylene glycol dimethacrylate as the cross-linker in a ratio of 1:10:80. The extracted analytes were quantitively determined using a newly developed and validated ultrahigh-performance liquid chromatography tandem mass spectrometry method, where the limits of detection and quantitation were 0.91 and 1.81 ng mL-1, respectively, fulfilling the minimum required performance limit decided on by the World Anti-Doping Agency. The mean percentage extraction recoveries for 3'-hydroxystanozolol, 4β-hydroxystanozolol, and 16β-hydroxystanozolol are 97.80% ± 13.80, 83.16% ± 7.50, and 69.98% ± 2.02, respectively. As such, the developed DMISPE can serve as an efficient cost-effective tool for doping and regulatory agencies for simultaneous clean-up of the stanozolol metabolites prior to their quantification.

Keywords: Bulk polymerization; Molecularly imprinted polymer; Solid-phase extraction; Stanozolol metabolites; UHPLC-MS/MS.

MeSH terms

  • Anabolic Agents / metabolism
  • Anabolic Agents / urine
  • Chromatography, High Pressure Liquid / methods
  • Doping in Sports* / prevention & control
  • Humans
  • Limit of Detection*
  • Molecular Imprinting / methods
  • Molecularly Imprinted Polymers* / chemistry
  • Solid Phase Extraction* / methods
  • Stanozolol* / urine
  • Substance Abuse Detection / methods
  • Tandem Mass Spectrometry / methods