Survey of nutritional supplements for selected illegal anabolic steroids and ephedrine using LC-MS/MS and GC-MS methods, respectively

Abstract

Several studies have highlighted that nutritional supplements may contain undeclared substances that are banned by the International Olympic Committee (IOC)/World Anti-Doping Agency (WADA). This paper describes a qualitative liquid chromatography coupled with tandem mass spectrometry detection (LC-MS/MS) method to detect anabolic androgenic steroids (4-androsten-3,17-dion, 4-oestren-3,17-dion, 5α-androsten-17β-ol-3-one, boldenone, nandrolone, nandrolone decanoate, testosterone, and testosterone decanoate) and ephedrine in food supplements. The products are dissolved in methanol and analysed by gas chromatography-mass spectrometry (GC-MS). The methanolic solution was added to testosterone-d₃, evaporated to dryness, mixed with NaOH and extracted with n-pentane:diethylether (9:1). LC-MS/MS analyses were performed in selected reaction monitoring (SRM) on an ion-trap equipped with an atmospheric pressure chemical ionization (APCI) probe operating in positive-ion mode. The method was applied to 64 nutritional supplements. A total of 12.5% of the nutritional supplements analysed contained banned substances not declared on the label (anabolic steroids and ephedrine). Detection limits were in the range 1–25 ng g⁻¹.

Keywords:

• Nutritional supplements
• anabolic steroids
• anabolic steroid esters
• prohormones
• liquid chromatography coupled with tandem mass spectrometry detection (LC-MS/MS) analysis