Identification of the small research tetra peptide Epitalon, assumed to be a potential treatment for cancer, old age and Retinitis Pigmentosa in two illegal pharmaceutical preparations Celine Vanhee,* Goedele Moens, Els Van Hoeck, Eric Deconinck and Jacques O. De Beer Keywords: biopharmaceuticals; LC-MS(n); geroprotective agent; anti-cancer agent Introduction SFFC (Spurious/falsely-labelled/falsified/counterfeit) medicines are a growing problem worldwide. While the problem in the develop- ing countries concerns principally essential medicines like antibiotics, [1] HIV medication, [2] and anti-malaria products, [3] the problems in the industrialized world concerns mostly drugs for erectile dysfunction, anabolic hormones, and slimming products. Although occasionally counterfeited antibiotics, insulin, and inter- feron are also intercepted by the customs or controlling agencies. [4] Moreover, there is also a growing market for non-approved drugs, including peptides and proteins, which are being made available to the public before entering or completing clinical trials. [5] Global actions are taken against criminal entities to curtail the trafficking and distribution of these substances. The growing threat of these products in Europe is mainly due to the extension of the Internet, [6–8] where about 50% of the medicines sold through Inter- net sites, disclosing their identity, are estimated to belong to this category. [9] In order to protect the costumer against such malice, a European directive was put in place in 2011. [10] A new logo for online purchases has to be implemented by member states by July 2015. This logo will allow patients and consumers to identify autho- rized online pharmacies providing authentic, authorized medicines. [10] At the request of the Federal Agency for Medicines and Health Products (FAMHP), unknown pharmaceutical preparations suspected to contain SFFC drugs, including protein and peptide drugs, are regularly subjected to analysis in the laboratory. Although almost all of the proteins and peptides analyzed cover potential doping agents, [11] the SFFC version of insulin, Botuli- num toxin, human interferon 1 α, nasal sprays containing the neuropeptide oxytocin, human serum albumin, and the potential toxic skin tanning peptide Melanotan II, were also found recently. In the last year, we have also received two different suspected illegal samples that contained a substance with a mass of approxi- mately 390 Da, corresponding to a peptide with sequence AEDG. The researchers who generated this peptide, termed Epitalon, claim that administration of this molecule had an inhibitory effect of the on the development of spontaneous tumours in mice, has geroprotective actions and intranasal administration increases neu- ronal activity. [12] Epitalon is currently also being studied in clinical trials for the treatment of Retinitis Pigmentosa. [13] Although all these studies show the plethora of potential beneficial effects of Epitalon, the peptide is still under pre-clinical and clinical development and thus has not been approved for any therapeutic and/or prophylac- tic use by any government health authority in Europe or the USA. Nevertheless, the samples seized by the Belgian health authorities clearly demonstrate that not only putative doping agents but also more and more essential biopharmaceuticals can be found through the internet or on the European black market. [9] Experimental Chemicals and reagents Acetonitrile was ultra liquid chromatography-mass spectrometry (ULC-MS) grade and formic acid, ethanol, and acetic acid were ana- lytical grade. All reagents were purchased from Biosolve (Valkenswaard, the Netherlands). Water was obtained using a milliQ-Gradient A10 system (Millipore, Billerica, MA, USA). A refer- ence standard of the peptides EADG and AED were custom synthe- sized to a respective purity of 88.1% and 97.8% and purchased from Thermo Scientific (Sunnyvale, CA, USA). * Correspondence to: Celine Vanhee, Division of Food, Medicines and Consumer Safety, Medicinal Products Section, Scientific Institute of Public Health (WIV-ISP), J. Wytsmansstraat14, B-1050 Brussels, Belgium. E-mail: Celine.Vanhee@wiv-isp.be Division of Food, Medicines and Consumer Safety, Medicinal Products Section, Scientific Institute of Public Health (WIV-ISP), J. Wytsmansstraat14, B-1050, Brussels, Belgium Drug Test. Analysis (2014) Copyright © 2014 John Wiley & Sons, Ltd. Correspondence case report Drug Testing and Analysis Received: 29 September 2014 Revised: 3 December 2014 Accepted: 5 December 2014 Published online in Wiley Online Library (www.drugtestinganalysis.com) DOI 10.1002/dta.1771