The effect of stock-culture period on the sensitivity of theLemna bioassay to four different phenolic substances was evaluated in this study. The sensitivity of the bioassay interacted with the stock-culture period of either 11, 14, or 18 days. After a seven-day testp-hydroxybenzoic acid (HBA), vanillic acid (VA),trans-cinnamic acid (CA), and ferulic acid (FA) with a concentration of 0.05 mM significantly reduced the number of fronds and the dry weight compared with the untreated control when the stock-culture period had been exactly 14 days. The sensitivity after the shorter (11 days) or longer (18 days) stock-culture period was reduced, and the differences in the dry weight to the untreated control were not significant after a stock culture period of 18 days. The two higher concentrations (0.1 and 0.25 mM) showed stronger inhibition. A comparison of the inhibition at 0.05 mM revealed that the stock-culture period affected the relative toxicity of the four phenolic substances. Since the pH increased in the stock-culture flasks during the 18-day period from 6.25 to 7.9, we hypothesize that differences in theLemna assay can be at least partly attributed to a pH effect, possibly in combination with a relative nutrient deficiency. We recommend a standardization of the stock-culture period to 14 days to improve the sensitivity of theLemna bioassay.