In Vitro Androgen Bioassays as a Detection Method for Designer Androgens

Abstract

Androgens are the class of sex steroids responsible for male sexual characteristics, including increased muscle mass and decreased fat mass. Illicit use of androgen doping can be an attractive option for those looking to enhance sporting performance and/or physical appearance. The use of in vitro bioassays to detect androgens, especially designer or proandrogens, is becoming increasingly important in combating androgen doping associated with nutritional supplements. The nutritional sports supplement market has grown rapidly throughout the past decade. Many of these supplements contain androgens, designer androgens or proandrogens. Many designer or proandrogens cannot be detected by the standard highly-sensitive screening methods such as gas chromatography-mass spectrometry because their chemical structure is unknown. However, in vitro androgen bioassays can detect designer and proandrogens as these assays are not reliant on knowing the chemical structure but instead are based on androgen receptor activation. For these reasons, it may be advantageous to use routine androgen bioassay screening of nutraceutical samples to help curb the increasing problem of androgen doping.

Current Status and Bioanalytical Challenges in The Detection of Unknown Anabolic Androgenic Steroids in Doping Control Analysis

“…Several bioassays based on the expression of AR (mainly mammalian cellbased bioas says and yeast cellbased bioassays) in the pres ence of AAS have been developed and recently reviewed [70]. The sensitivity of bioassays based on AR activities mainly depends on the andro genic activity of the AAS.…”

Annual banned‐substance review: analytical approaches in human sports drug testing

“…[53] One of the main advantages of this approach is the assay's capability to indicate the presence of one or more substances able to bind to the androgen receptor, even if the structures and compositions of the substrates are unknown to the analyst. [53] One of the main advantages of this approach is the assay's capability to indicate the presence of one or more substances able to bind to the androgen receptor, even if the structures and compositions of the substrates are unknown to the analyst.…”

“…The utility of androgen receptor-based bioassays to probe for the presence of AAS and other non-steroidal anabolic agents in dietary products (with and without additional mass spectrometric measurement) has been demonstrated with various applications and reports in the past. [53] One of the main advantages of this approach is the assay's capability to indicate the presence of one or more substances able to bind to the androgen receptor, even if the structures and compositions of the substrates are unknown to the analyst. Moreover, the bioassay will provide information on the sum of androgen receptor activation.…”

Direct analysis in real time ‐ high resolution mass spectrometry (DART‐HRMS): a high throughput strategy for identification and quantification of anabolic steroid esters

“…[12][13][14][15][16] Recently, androgen receptor-based bioassays have proved their ability to detect the presence of anabolic steroids in dietary products. [17] In biological matrices, such as hair or blood, the identification of steroid esters, one of the chemical forms under which the steroids are administered, is considered as one of the most efficient analytical strategies to confirm the use of such naturally occurring steroids. Nevertheless, their detection at trace levels constitutes a great challenge requiring extensive sample preparation and last generation analytical instruments such as liquid chromatography coupled to mass spectrometry (MSn or high resolution mass spectrometry (HRMS)).…”