Detection of the protease bacillolysin in doping-control urine samples.

Industrial, pharmaceutical and biochemical applications for proteases have a long history and proteolytic processes have been examined and understood for decades.[1] In contrast, the adulteration of urine samples in sports drug testing may represent a more urgent and recent phenomenon in the detection of protein-based prohibited substances.[2 – 5] Nowadays, enormous amounts of various peptidases are produced in bacteria in order to customize every proteolytic problem in industrial or biochemical implementation. Universal mixtures of proteases for optimal digestion under different conditions (such as pH, temperature and time of activity) are commercially available allowing proteolysis under all kinds of external conditions. Cheating sportsmen thus have a large variety of effective proteases to choose from, which needs to be taken into account when conducting doping analysis of proteinbased performance-enhancing agents. This case report presents the sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography-mass spectrometry (LCMS) approaches for the detection of manipulation using a protease in doping-control urine samples.