Analysis and evaluation of black market nasal sprays containing peptides and proteins

Code: 242B02CR

Chapter S2 of WADA’s Prohibited List 2024 (“Peptide hormones, growth factors, related substances, and mimetics”) lists under sub-chapters 2 and 3 “Peptide hormones and their releasing factors” and “Growth factors and growth factor modulators” as prohibited substances. Chapter S4 “Hormone and metabolic modulators”, sub-chapter 3 (“Agents preventing activin repetor IIB activation) additionally mentions “Activin receptor IIB competitors” (e.g. ACE-031) and “Myostatin inhibitors” (e.g. follistatin, myostatin propeptide) as prohibited proteins. Most of these compounds have not been pharmaceutically approved but are available on the black market. In the past, only studies on injectable black market peptides and proteins were performed. They led to the identification of unknown compounds with doping relevance and revealed the presence of “tags”, which made them clearly distinguishable from the body's own proteins (e.g. follistatin with His-tags, myostatin propeptide with GST-tags). Recently, an increasing amount of nasal sprays containing growth promoting peptides and proteins (e.g. AOD-9694, ACE-031, CJC-1295, Follistatin 344, GDF-8) appeared on the black market. So far, no comprehensive study on these products has been performed. Hence, it is unclear whether the compounds contained in nasal sprays are detectable with the current methods or not. For example, some might contain “tag-free” proteins that pass through mucous membranes more easily but are undetectable using existing methods. Within the scope of this project, ca. 50 peptide/protein-based nasal sprays will be purchased on the black market and analyzed with high resolution mass spectrometry and gel-electrophoresis. Subsequently, their detectability with the currently applied methods will be evaluated. In case of unknown compounds, top-down and bottom-up proteomics will be applied for identification. For compounds which cannot be detected with the current methods, alternative strategies will be proposed.