Fluorescence lifetime measurement in the time domain requires excitation from a well separated single bunch using synchrotron light sources. In the colliding mode of the Beijing Electron Positron Collider II (BEPCII), a hybrid filling pattern was realized such that a single bunch was placed in the middle of a large gap between two multi-bunch groups. Detection of fluorescence lifetime, based on the excitation of the light pulse from this designated single-bunch, was established at Beamline 4B8 of the Beijing Synchrotron Radiation Facility (BSRF). The timing signal of the BEPCII was utilized as a trigger to gate this fluorescence event. L-Tryptophan amino acid, a known lifetime standard, was selected to assess the lifetime measurement performance. The measured lifetime was consistent in both colliding and single-bunch mode with the time resolution down to 450 ps. Moreover, both the bunch purity and the fine structure of the hybrid filling pattern were characterized.