Hydrogels are an important class of biomaterials that can absorb large quantities of water. In this study, changes in hydration of natural hydrogels (agar, chitosan, gelatin, starch, and blends of each with chitosan) during storage and rehydration were studied by using near‐infrared hyperspectral imaging (NIR‐HSI). Moisture content was calculated based on changes in sample weight during hydration. The NIR‐HSI data were acquired by using a push‐broom system operating in diffuse reflectance in the wavelength range 943 to 1650 nm. A novel synthesis method was developed to enable common preparation of each hydrogel. Mean spectra obtained from the hyperspectral images were analyzed, and predictive models for moisture content were developed by using partial least squares regression. Models were compared in predictive performance by using an independent validation set of data. The optimal model in predictive performance was a 1 latent variable partial least squares regression model developed on second derivative and mean centered pseudo‐absorbance data in the wavelength range 943 to 1272 nm. This model was applied to pixel spectra from samples in the validation set to inspect spatial variations during dehydration and rehydration. Challenges associated with NIR‐HSI of hydrogels with a large variation in moisture content are discussed.
