AbstractA molecularly imprinted polymer (MIP) is a biomimetic material that can be used as a biochemical sensing element. We studied the steady‐state and time‐resolved fluorescence and fluorescence anisotropy of anthracene‐imprinted polyurethane. We compared MIPs with imprinted analytes present, MIPs with the imprinted analytes extracted, MIPs with rebound analytes, non‐imprinted control polymers (non‐MIPs) and non‐MIPs bound with analytes to understand MIP's binding behaviour. MIPs and non‐MIPs had similar steady‐state fluorescence anisotropy in the range 0.11–0.24. Anthracene rebound in MIPs and non‐MIPs had a fluorescence lifetime of τ = 0.64 ns and a rotational correlation time of ϕF = 1.2–1.5 ns, both of which were shorter than that of MIPs with imprinted analytes present (τ = 2.03 ns and ϕF = 2.7 ns). The steady‐state anisotropy of polymer solutions increased exponentially with polymerization time and might be used to characterize the polymerization extent in situ. Copyright © 2005 John Wiley & Sons, Ltd.