Human calcitonin is a 32-residue peptide hormone that binds to the calcitonin receptor (CTR) and is involved in calcium regulation. The amino acid sequence displays a hydrophilic central segment flanked by hydrophobic C- and N-terminal regions with a net charge of zero at neutral pH. This makes the molecule amphiphilic and conformationally flexible, and different CTR variants preferentially recognize different structural conformations of calcitonin. The peptide is secreted from the thyroid gland and is overproduced in some forms of thyroid cancer and can then form cell-toxic aggregates. Characterizing the structural properties of calcitonin under different conditions is, therefore, important for understanding its receptor-binding and self-aggregation properties. Here, we used circular dichroism (CD) spectroscopy to monitor the secondary structure of human calcitonin in different environments. Calcitonin monomers were found to display a random coil structure with a significant amount of PPII-helix components in phosphate buffer, pH 7.3, at physiological temperatures. When agitated, the peptide formed soluble aggregates over time with mainly an antiparallel β-sheet secondary structure. In the presence of micelles of differently charged surfactants, monomeric calcitonin formed a pure α-helix structure with cationic CTAB, a combination of α-helix and β-sheet with anionic SDS and with zwitterionic SB3-14, and remained mainly random coil with noncharged DDM. Thus, the charge of the surfactant headgroup was found to be an important parameter for calcitonin’s interactions with membrane-mimicking micelles. Similar but not identical interactions with the surfactants were observed under the oxidizing and reducing conditions.