Betula and Pinus pollen, which are dispersed in natural surface waters, release biologically active compounds into the water bodies. This study aims to evaluate variations in the distribution and composition of phenolic compounds in suspended particles in natural water bodies during pollen spreading. Samples taken from water bodies of different trophic levels were analyzed by microscopy, UV/VIS spectroscopy, HPTLC, and HPLC/DAD. The study revealed that the total phenolic content in water-suspended particles varied from 3.0 mg/g to 11.0 mg/g during Betula and Pinus pollen spreading. It was also observed that the surface water of dystrophic natural lakes had a higher content of phenolic compounds than the eutrophic, hypereutrophic, and mesotrophic water bodies. Chlorogenic, trans-ferulic, vanillin, and 3,4-dihydroxybenzoic acids were frequently detected in the surface water samples. Experimental measurements have shown variations in the release of phenolic compounds from Betula pollen into water (p < 0.05). After the exhibition of pollen, the distilled water predominantly contained bioactive chlorogenic acid. Further in situ investigations are necessary to gain a more comprehensive understanding of the function of phenolic compounds in aquatic ecosystems. The exploration of the release of bioactive compounds from pollen could provide valuable insights into the potential nutritional value of pollen as a nutrient source for aquaculture.