An LC–MS screening method with library identification for the detection of steroidsin dietary supplements

Abstract

For many years anabolic-androgenic steroids (AAS) are by far the most frequently detected pharmacological substances in doping control. In order to improve their performances, professional sportsmen are often tempted to take dietary supplements. However, due to the frequent and widespread occurrence of contaminated supplements, the use of such products is not without risk for the athletes involved. In order to minimize the chances of an unattended positive doping test or serious health problems, fast and reliable screening methods for the detection of anabolic steroids in dietary supplements are needed. A general screening procedure requires the fast and unambiguous detection of a large range of steroids. Gas chromatography-mass spectrometry (GC-MS) has been used intensively in the detection of doping substances for the past 40 years. Over time, many laboratories have delivered spectra to be included in standard reference databases, one of which is maintained by the National Institute of Standards and Technology (NIST) (Gaithersburg, MD, USA). In recent years, however, liquid chromatography coupled to mass spectrometry (LC-MS) has gained popularity. Unfortunately, existing GC-MS libraries are not applicable to LC-MS analysis. In the present study, a new mass spectral library of 88 steroids was developed, along with a fast UPLC-MS method. For the construction of this mass spectral library, three different mass spectra were measured for each steroid, with a sample cone voltage of 30, 60 and 100 V, respectively. This method was then successfully tested on contaminated dietary supplements which had previously been tested by means of a targeted LC-MS/MS method. Overall, the library search was shown to identify the same compounds as the MRM method.

Determination of anabolic agents in dietary supplements by liquid chromatography-high-resolution mass spectrometry

“…The analysis of food supplements requires sample pretreatment in order to have them in the right physical state for analysis, to reduce interferences present in the matrix and to detect low concentrations of analytes. The techniques used till now for these purposes involved a dissolution in a solvent (Van Poucke et al 2007;Doué et al 2014;Krug et al 2014) followed sometimes by liquidliquid extraction (LLE) Baume et al 2006;Martello et al 2007;Strano-Rossi et al Forthcoming), otherwise by SPE (Becue et al 2011) or a filtration (Cho et al 2014). The subsequent detection step of AASs is mostly performed by GC-MS (Kamber et al 2001;Delbeke et al 2002;Geyer et al 2004;Baume et al *Corresponding author.…”

Adulteration of Dietary Supplements by the Illegal Addition of Synthetic Drugs: A Review

“…Different studies recently available in the literature have shown the presence of anabolic steroids and designer drugs in dietary supplements, such as androstenedione, 5‐androsten‐3β‐ol‐17‐one (dehydroepiandrosterone; DHEA), methandienone, testosterone esters, androst‐4‐ene‐3β‐17β‐diol, boldenone, among others (Geyer and others ; Becue and others ; Aqai and others ; Odoardi and others ). Table shows a compilation of studies reporting the presence of illegal substances in dietary supplements.…”

“…Additionally, the use of optimized sample extraction and clean‐up protocols has been suggested by different authors, including the use of liquid‐liquid extraction (Geyer and others ; Schramek and others ; Strano‐Rossi and others ), solid‐phase extraction (SPE) (Stepan and others ; Peters and others ; Becue and others ), and QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure (Vaclavik and others ). When analyzing 88 steroid compounds in PFS using ultra‐performance liquid chromatography‐MS (UPLC‐MS), Becue and others () concluded that a clean‐up procedure using a SPE NH 2 ‐column was necessary for improved sensitivity and selectivity, since it allowed for a substantial reduction of background noise by retention of polar matrix effects. Stepan and others () employed dispersive SPE using primary secondary amine (PSA) as a sorbent to effectively remove polar components such as sugars, 4‐hydroxy‐2‐methoxycinnamaldehyde and partially remove fatty acids and vanillin.…”

Development of an ultra‐high performance liquid chromatography–tandem mass spectrometry method for the determination of anabolic steroids currently available on the black market in the Czech Republic and Slovakia

“…The increasing distribution and misuse of black market products urgently requires comprehensive analytical procedures for their identification. Several analytical methods published in recent years have focused on the analysis of counterfeit AAS products primarily using liquid chromatography (LC) and gas chromatography (GC), often coupled with mass spectrometry (MS) . Although studies have focused on testing the quality of AAS products, quantitative analysis is necessary to avoid underestimating the counterfeit rate of AAS products .…”

“…Several analytical methods published in recent years have focused on the analysis of counterfeit AAS products primarily using liquid chromatography (LC) and gas chromatography (GC), often coupled with mass spectrometry (MS). 3,[11][12][13][14][15][16] Although studies have focused on testing the quality of AAS products, 16,17 quantitative analysis is necessary to avoid underestimating the counterfeit rate of AAS products. 10 different dietary supplements and concluded that 60% of the samples contained at least one AAS.…”