Non-isocyanate polyurethanes (NIPUs) with varying content of secondary amino groups along their chain were studied with respect to water absorption and plasticization. Secondary amino groups promote water uptake. Water sorption isotherms in the water activity range 0–0.97 are discussed in terms of various sorption models. Secondary amino groups act as additional hydration sites, increasing monolayer capacity. The red shift of the IR band assigned to the carbonyl of the urethane and the simultaneous blue shift of the urethane N–H bending band show cleavage of the polymer-polymer hydrogen bonds upon water uptake, due to strong interactions between water molecules and hydroxyurethanes constituting the primary hydration sites. A decrease in glass transition temperature (Tg) is observed with increasing water content. The effect is discussed in terms of plasticization and slaving mechanisms, while a peculiar decrease of Tg at very low hydrations is attributed to a mechanism not following common mixing laws. Display omitted •Secondary amino groups along NIPU chain contour act as secondary hydration sites.•Water absorption of NIPUs can be tailored by controlling polar groups concentration.•At low hydrations standard mixing models fail to describe Tg decrease.•This is attributed to scission of inter-segment hydrogen bonds by traces of water.