Twelve biochemical parameters were measured in serum blood samples from rainbow trout (Oncorhynchus mykiss) maintained under controlled conditions. Forty-five samples were taken every 3 months (T1–T4) over the course of one year to define baseline values. The effect of fish total weight (TW) was also evaluated. Principal component analysis showed a clear separation between T4 and T1, and T2 and T3, indicating an increase in certain biochemical parameters with weight. Linear regression analysis showed how TW significantly explained 11–67% of the variability observed for nine parameters out of 12. Pearson’s correlation matrix showed a significant positive correlation (p p O. mykiss, indicating that fish weight is an important factor for interpreting the blood biochemical profile.