In this work, we evaluated the effect of harmful ecotoxins, 4,5,6-trichloroguaicol (4,5,6-TCG) and tetrachloroguaiacol (TeCG), on the oxidation of the fluorescent probe dihydrorhodamine 123, the content of free phenols and the level of the total, oxidized and reduced glutathione in the leaves of reed canary grass (Phalaris arudinacea). Furthermore, the effect on the activity of guaiacol peroxidase and glutathione S-transferase was investigated. Both 4,5,6-TCG and TeCG increased the activity of guaiacol peroxidase and glutathione S-transferase, they also elevated the content of free phenols and the level of the total glutathione. A stronger effect was exerted by tetrachloroguaiacol, which strongly increased the level of the total glutathione and the content of free phenols on the 3rd and 6th day of the experiment. The activity of glutathione S-transferase was more intensively induced by trichloroguaiacol. Both 4,5,6-TCG and TeCG oxidized dihydrorodamine 123 and the effect was stronger in the presence of magnesium ions.