Endogenous inhibitors have a crucial effect on the success of sugar beet (Beta vulgaris) seed germination. The strength of this effect changes during seed maturation, and washing away these inhibitors can facilitate germination. Investigation of various washing factors such as water-to-seed ratio, stirring time, and speed of stirring on a mixed-maturity lot of sugar beet seeds revealed the significant effect of the interaction of all factors. Seeds with different maturity classes, from the same lot, were washed using procedures determined from the first experiment. Statistical analysis of the germination data of the washed seeds showed a significant difference in mean germination time depending on maturity class. Washing seeds of different maturity classes with 40 mL of water per gram of seed improved the germination performance of maturity classes 2 and 5, compared with 20 mL water per gram of seed. The UV absorbance of inhibitors washed from seeds differed between maturity classes. The change in seed moisture content during the washing process is another factor altered by the extent of seed development, and this variation in water uptake may reflect the pericarp structure of seed in different maturity classes. The findings in this manuscript highlight the importance of optimizing the pre-treatment procedures in accordance with the individual seed lot.