This study investigates the potential use of attenuated total internal reflection-Fourier transform infrared (ATR-FT-IR) imaging, a hyperspectral imaging modality, to investigate molecular level trends in the interaction of water with polymeric surfaces of varying hydrophobicity. The hydrophobicity of two categories of polymeric biomaterials is characterised using contact angle (CA) measurements and their relationship with the band area of the OH stretching νs vibration of water over time is presented. This is supported with correlations between CA data and single wavenumber intensity values (univariate analysis). Multivariate analysis of the spectra captured at the OH stretch for all polymers is carried out using principal component analysis to study the spatial variation in the interaction between the polymeric surfaces and water. Finally, a comparison between the univariate and multivariate strategies is presented to understand the interaction between polymeric biomaterials and water.