Study of interfacial processes between incorporated biomolecules and hydrogel matrix is of primary importance for the application of synthetic hydrogels as biomaterials, sensors, etc. Their hydrophilic nature, however, hampers fat-soluble biomolecule incorporation. We synthesized N-isopropylacrylamide (NIPAAm)- and Acrylamide (AA)-based hydrogels with hydrophilic-lipophilic balance that under specific conditions permit homogeneous incorporation of provitamins D3(7-dehydrocholesterol) and D2(ergosterol) dissolved in ethanol. Similarity of provitamins D photoisomerization in such hydrogel matrices with the photoreaction in ethanol shows great potential of the novel material for personal dosimetry of biologically active ‘antirachitic’ UV radiation.