Characterization of the unfolding kinetics of G-quadruplexes (G4s) under physiological condition is the key to a better understanding of the biological function of G4s and is important for biomedical research and material design. Of interest is that that slight variations of human telomeric sequences can form different types of G4 structures. It is important to examine whether there is a correlation between thermal stability and unfolding kinetics of these various G4 structures. Here we report systematic analyses of the unfolding kinetics of a variety of telomeric G4 structures by monitoring the real-time imino proton NMR spectra of DNA hybridization and hydrogen-deuterium exchange (HDX). From aspect of thermodynamic, we measured the melting temperature (Tm) and determined the Gibbs free energy (∆G) based on the differential scanning calorimetry (DSC) of these various G4 structures. Both of them showed slight structure-dependence, except the Tm of parallel G4 structure is ~10 oC higher than that of nonparallel G4 structure. We further used imino proton NMR spectra to monitor unfolding kinetics of these various telomeric G4 structures as a function of time based on hybridization DNA and hydrogen-deuterium exchange (HDX) experiments. The hybridization results showed that the decay times of different imino proton signals for each telomeric G4 structure are quite similar, which are also consistent with the time constant obtained from HDX measurements. It is suggested that G4 unfolding is rate-determining step and our NMR measurements can approach the intrinsic unfolding kinetic rate constant. The key finding is that the unfolding times of these various G4 structures are quite different and shows no correlation between thermal stability and unfolding kinetics. Our results raised an issue that kinetics of the folding and unfolding are more relevant for better understanding of biological function of G4 structure.