Denitrification in floodplain soils is one key process that determines the buffering capacity of riparian zones in terms of diffuse nitrate pollution. One widely used approach to measure the denitrification potential is the acetylene inhibition technique that requires fresh soil samples. We conducted experiments with air-dried soils using a time series analysis to determine the optimal rewetting period. Thus, air-dried soil samples from six different floodplain areas in Germany were rewetted for 1 to 13days to 100% water-filled pore space. We analyzed nitrogen accumulated as N2O in the top of anaerobic flasks with and without acetylene by gas chromatography after four hours of incubation. We observed an overall optimal rewetting of at least seven days for complete denitrification. We also saw the strong influence of pH and field capacity on the denitrification product ratio; in soils with pH < 7, we hardly assumed complete denitrification, whereas the treatments with pH > 7 achieved stable values after seven days of rewetting. This advanced method provides the opportunity to carry out campaigns with large soil sample sizes on the landscape scale, as samples can be stored dry until measurements are taken.