Polyphenols are chemicals that primarily inhibit the growth of various autotrophic organisms. The presence of these metabolites greatly boosts the ecological dominance of eg. Cladophora, which creates large surface mats. The main goal of our work was to quantify the phenol and polyphenols (allelopathic substances) secreted by the macroalgae as a result of exposure to biotic stress caused by competition. The research was carried out on the Cladophora glomerata biomass collected from two freshwater ecosystems located in Wielkopolska Region (Poland, Fig. 1): Oporzynskie Lake (N52° 55′, E 17° 9′) and Nielba River (N52˚48′, E 17° 12′). Seasonal variability (May-October 2015) in the properties of C. glomerata mats in the river and lake ecosystem was also analyzed in relation to the physicochemical parameters of water. In addition, the content of pigments in the analyzed biomass was determined during the appearance of algae mats in water reservoirs. Biomass extraction was performed to determine the phenolic and carotenoid contents (chlorophyll and carotenoids) by using two extraction methods: microwave-assisted extraction (MAE) and supercritical fluid extraction (SFE). After isolation of the phenols from the thalli (mats) and the habitat, they were analyzed using the Folin-Ciocalteu method with some modifications, while the pigment content (chlorophyll and carotenoids) was evaluated by the spectrophotometric method Liechtenthaler (1987) with some variations. Analysis of the content of these components in algae extracts indicates that the tendency of changes in their contents was similar or the same. Growth and decrease of phenolic content (Cladophora T MAE and Cladophora T SFE) and pigments (MAE chlorophyll, SFE and MAE carotenoids) at the same time were independent of the insulation method used. The mats formed by C. glomerata on the surface of Lake Oporzyńskie were more stable and larger surface area than those on the Nielba River, which could explain differences in polyphenol concentrations in these two aquatic ecosystems. The results suggest a reduction in the secretion of phenolic compounds with an aging population of algae.