Abstract: In a method for isolating analytes of a first analyte class, which are extracellular vesicles and/or circulating tumor cells, and analytes of a second analyte class, which are cell-free nucleic acids, from the same sample volume of a biological sample in a centrifugal microfluidic system, the sample volume is guided into a first isolation chamber of a fluidics module containing a first isolation structure so that the analytes of the first analyte class are retained by the first isolation structure while the analytes of the second analyte class are not retained and pass through the first isolation structure as part of a residual liquid, which is passed into a second isolation chamber of the fluidics module containing a second isolation structure so that the analytes of the second analyte class are retained by the second isolation structure. The analytes of the first analyte class and the second analyte class are separated from the respective isolation structure to provide the analytes for subsequent analysis.