Fluorescence lifetimes have been examined of 1, N6-ethenoadenosine (epsilon Ado), 1-methyl-ethenoadenosine (m1 epsilon Ado+), 9-methyl-ethenoadenosine (m9 epsilon Ado+), N1-deazaethenoadenosine (N1-deaza-epsilon Ado, and 9-methyl-N1-deazaethenoadenosine (m9N1-deaza epsilon Ado) at various pH's in the 7.5 approximately 1.5 region and at 20 degrees C. From an analysis we have reached the following conclusions. (1) The observed fluorescence of epsilon Ado is caused by the unprotonated, neutral, excited species epsilon Ado not not only at pH 7 but also at pH 2. (2) pKa of the excited epsilon Ado is lower than that of the ground state epsilon Ado. (3) The rate constant of protonation in the excited state is about 10(10) sec-1 M-1.