The study addressed the effect of the structure of silicone polyethers on selected functional properties of cotton fabric rinsed in conditioners containing the additives under study. Fabric softener formulations containing two comb-structured compounds (PEG/PPG-14/0 Dimethicone and PEG/PPG-20/20 Dimethicone) and one block-structured compound (Bis-PEG/PPG-20/20 Dimethicone) were developed. Cotton fabric rinsed in conditioners containing silicone glycols was not found to be affected by yellowing. However, differences were noted in the softening ability and re-wettability of rinsed fabrics due to diverse structures of the additives used. The most desirable soft hand effect was observed after cotton rinsing in fabric softeners containing the block-structured compound Bis-PEG/PPG-20/20 Dimethicone. In contrast, the highest fabric re-wettability was shown for the conditioner enriched with a comb-structured compound (PEG/PPG-20/20 Dimethicone). The study results demonstrate that the prototypical fabric softeners containing silicone derivatives have a potential to provide quality characteristic required by consumers of this product group.