To make the use of MALDI-based mass spectrometry feasible for the fast analysis of oxosteroids, three new aromatic probes have been designed to be used simultaneously as derivatisation agents and MALDI matrices. This concept brings a number of benefits: the sample handling is reduced, the workflow is less time consuming allowing high throughput and the interferences caused by the MALDI matrix are avoided. Identification was successfully attained for all oxosteroids used in this study. As proof-of-concept, the identification of oxosteroids in urine was performed to evaluate the robustness of the new methodology. The oxosteroids 17-α-methyltestosterone, nandrolone, boldenone, 17-α-Trenbolone, fluoxymesterolone, mesterolone and bolasterone were identified in human urine at the minimum concentration level recommended by the world anti-doping agency, 2 ng/mL.
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