Dipalmitoylphosphatidylcholine (DPPC) demonstrated complex differential scanning calorimetry (DSC) thermal behavior. Transitions below 100°C showed variability in their thermotropic reversibility. An experimental design employing a DSC heat-cool-heat-cool-heat cycle and modulated DSC were used to gain insight into the DPPC's complex thermal nature. An annealing strategy was developed to reduce DPPC's thermotropic variability, moisture uptake rate, and rate variability. Samples annealed at 110°C for 5 min provided a reproducible, thermally reversible material. The annealed material also exhibited an 8-fold decrease in moisture sorption rate and a statistically significant (p = 0.0233) 100-fold decrease in water sorption rate variability compared to DPPC “as is.” An optimized validated stability-indicating high performance liquid chromatography with evaporative light scattering detection method was developed and showed no change in DPPC chemical stability under the annealing treatment conditions.