Dear Sir. Concerning your issue about the method of detecting EPO in biological samples using MAIIA monolith columns purification kit on a HRMS. To enhance the endurance performance the misuse of erythropoietin (EPO) is currently detected by isoelectric focusing (IEF) based on the profile of altered isoforms of EPO. Changes within the new WADA technical document allows for the detection of the different erythropoietins by using the SarcosylPAGE, a modified SDS-PAGE. The Sarcosyl-PAGE which separates by difference in the molecular mass of EPO isoforms introduces a number of advantages: clear evidence between endogenous and the new modified exogenous EPO even when applied in smaller quantities; extended detection time of exogenous EPO, and a more simplified analysis. For EPO routine analysis a protocol based on a new type of Sarcosyl precast gels was established. For the evaluation of this improved detection method blood and urine samples of a micro-dose study were investigated. Affinity purification was performed according to the manufacturer’s instructions (MAIIA AB). Briefly, 2 ml UPD-buffer was added to the urine samples, which were incubated for 10 min at RT and then heated on a simmering water bath for 10 min. After cooling, samples were diluted with 20 ml working sample buffer and filtered through Steriflip filters. Urines were passed through the monolith columns at 1 ml/min using a vacuum manifold at a pressure of -5 kPa. After washing, the isolated EPO was eluted with 40 μl desorption buffer into 4 μl adjustment buffer. The aliquots (20 µl) were at -20 °C until use. Serum (1-2 ml) was diluted 1:10 in working sample buffer, filtered through Steriflip filters (0.2 μm) and then affinity purified using the same columns and desorption buffers as for urine. I think the following below links and the attached file may help you in your analysis:

http://www.dyeagnostics.com/site/wp-content/uploads/2015/02/Application_Note_Improvements-for-EPO-detection_Schwenke_2015.pdf

https://www.ncbi.nlm.nih.gov/pubmed/24190107

https://www2.bc.edu/christopher-kenaley/bio3030/Schumacher.pdf

Thanks

Also you can see the below attached file: