on 02-28-201302:00 AM - edited on 10-15-202109:03 AM by AnalyteGuru
Application Note 585 Recombinant human erythropoietin (rHuEPO) is a 30–34 kDa glycoprotein and a common doping agent banned by equestrian and racing authorities. To differentiate between equine endogenous EPO and rHuEPOs, methods employing selected-reaction monitoring (SRM) or isoelectric focusing (IEF) with double-blotting have been developed. However, the short confirmation-time window of the compound in plasma continues to present a challenge for doping control laboratories in horse racing. In this study, a new sample preparation technique using small anti-EPO monolith membranes contained in disposable columns (anti-EPO monolith columns) was combined with nanoLC-MS/MS analysis on a benchtop high-resolution Thermo Scientific™ Orbitrap™-based mass spectrometer. The approach enabled the confirmation and quantitation of the rHuEPO target peptides in a single analytical run, with high specificity, sensitivity, and resolution.