Protein modification by aldehydes has been generally associated with aging and onset of various chronic and cardiovascular diseases. The characteristics of bovine serum albumin (BSA) incubated with three dien-aldehydes (trans, trans-2,4-heptadienal, trans, trans-2,4-nonadienal, trans, trans-2,4-decadienal) of different chain lengths at different concentrations were examined. The results were as follows: loss of the amino group and increased carbonyl value were indicative of BSA side chain damage by these three unsaturated aldehydes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis results showed that all the three aldehydes led to formation of BSA aggregation; the most significant effect was observed for heptadienal. Changes in intrinsic fluorescence and surface hydrophobic value of BSAs indicated that aldehydes modified protein structures of BSAs. Moreover, the three aldehydes used in this experiment caused BSAs to form yellowish-brown adducts and fluorescent lipofuscin. The heptadienal-BSA adducts exhibited increased ultraviolet–visible (UV-Vis) absorbance at 270-280 nm and 300-400 nm, similar to the two other aldehyde-BSA adducts. We also examined the correlation between the various oxidation parameters and the concentration of modifiers. Strong correlations were observed between formation of protein-bound carbonyls, the retention ratio of free amino content, maximum UV-Vis absorption value, and concentration of aldehydes. Finally, Principal component analysis (PCA) analysis was conducted on oxidation parameters and the comprehensive effect of these parameters on BSA modification. In general, greater BSA damage was observed when incubated with aldehydes with shorter chain length at higher concentration.