We obtained the chemical shifts of amide protons (NHs) in helical peptides at various temperatures and trifluoroethanol (TFE) concentrations using 2-dimensional NMR spectroscopy. These NH chemical shifts and their temperature dependence exhibited characteristic periodicity of 3-4 residues per cycle along the helix, where downfield shifted NHs showed larger temperature dependence. In an attempt to understand these observations, we focused on hydrogen bonding changes in the peptides and examined the validity of two possible explanations: (1) changes in intermolecular hydrogen bonding caused by differential solvation of backbone carbonyl groups by TFE, and (2) changes in intramolecular hydrogen bonding due to disproportionate variations in the hydrogen bonding within the peptide helix. Interestingly, the slowly exchanging NHs, which were on the hydrophobic side of the helix, showed consistently larger temperature dependences. This could not be explained by the differential solvation assumption, because the slowly exchanging NHs would become more labile if the preceding carbonyl groups were preferentially solvated by TFE. We suggest that the disproportionate changes in intramolecular hydrogen bonding better explain both the temperature dependence and the exchange behavior observed in this study.