Review on activity-based detection of doping substances and growth promotors in biological matrices: do bioassays deserve a place in control programs?

Background: Control programs such as anti-doping control and growth promotor residue surveillance programs are challenged by the emergence of designer drugs and the use of low-level drug cocktails. In order to cope with these challenges, the use of bioassays, measuring biological activity in a matrix, has been explored over the past two decades as a universal means to detect (combinations of) unknown drugs, regardless of their chemical structure.
Results: This review compiles the experience on the use of activity-based assays to detect doping substances and growth promotors in biological matrices of humans (athletes) or live animals (race and/or food-producing animals). The aim is to learn from the scientific progress, going from initial research to the recent revival of this topic. Bioassay improvements and remaining limitations are discussed, along with a rational evaluation of possible applications of bioassays in control programs at their current functionality. Limitations include the possible interference by endogenous compounds and the challenge to detect metabolically activated (pro-)drugs. Nevertheless, successful validation of bioassays has been achieved, ensuring robust, reliable and valid results.
Significance: We conclude by proposing three applications of bioassays that provide added-value to the current testing procedures: (i) characterization of compounds to provide indisputable proof of biological effects and to prioritize legislative (cf. expansion of bans) and research endeavors (cf. method development), (ii) bioassay-based screening of biological samples to direct intelligent sample storage, sample retesting and targeted athlete testing, (iii) bioassay-guided identification of drugs to overcome the challenges of suspicious peak selection, related to high-resolution techniques.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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