Peptide profiling of Internet-obtained Cerebrolysin using high performance liquid chromatography - electrospray ionization ion trap and ultra high performance liquid chromatography - ion mobility - quadrupole time of flight mass spectrometry

• Published in: Drug testing and analysis 2015-09, Vol.7 (9), p.835-842
• Main Authors: Gevaert, Bert, D'Hondt, Matthias, Bracke, Nathalie, Yao, Han, Wynendaele, Evelien, Vissers, Johannes Petrus Cornelis, De Cecco, Martin, Claereboudt, Jan, De Spiegeleer, Bart
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino Acids - chemistry; Amino Acids - supply & distribution; Animals; Cerebrolysin; Chromatography, High Pressure Liquid; cryptic peptides; high definition mass spectrometry; Internet; Internet-obtained medicines; ion mobility separation; ion mobility separation, Internet‐obtained medicines; ion trap mass spectrometry; Molecular Sequence Data; Neuroprotective Agents - chemistry; Neuroprotective Agents - supply & distribution; Peptides - analysis; Proteins - analysis; Spectrometry, Mass, Electrospray Ionization; Swine
• ISSN: 1942-7603; 1942-7611
• DOI: 10.1002/dta.1817

Cerebrolysin, a parenteral peptide preparation produced by controlled digestion of porcine brain proteins, is an approved nootropic medicine in some countries. However, it is also easily and globally available on the Internet. Nevertheless, until now, its exact chemical composition was unknown. Using high performance liquid chromatography (HPLC) coupled to ion trap and ultra high performance liquid chromatography (UHPLC) coupled to quadrupole‐ion mobility‐time‐of‐flight mass spectrometry (Q‐IM‐TOF MS), combined with UniProt pig protein database search and PEAKS de novo sequencing, we identified 638 unique peptides in an Internet‐obtained Cerebrolysin sample. The main components in this sample originate from tubulin alpha‐ and beta‐chain, actin, and myelin basic protein. No fragments of known neurotrophic factors like glial cell‐derived neurotrophic factor (GDNF), neurotrophin nerve growth factor (NGF), brain‐derived neurotrophic factor (BDNF), and ciliary neurotrophic factor (CNTF) were found, suggesting that the activities reported in the literature are likely the result of new, hitherto unknown cryptic peptides with nootropic properties. Copyright © 2015 John Wiley & Sons, Ltd. Cerebrolysin, a parenteral pig‐brain‐derived product is easily available on the Internet. Here, we present for the first time its peptide composition. The peptides were identified by HPLC ion trap MS and UHPLC ion mobility‐quadrupole‐time of flight MS, using both database and de novo approaches.