Colloidal gold solutions with nanostars and nanospheres as well as Klarite™ gold and gold-copper bimetallic substrates were used for SERS analysis of aniline and nitroaniline isomers to investigate their chemisorption phenomena. Computational modeling based on Density Functional Theory (DFT) was used in conjunction with the SERS analysis to study the adsorption behaviors of the analytes on metal surfaces. Gold nanospheres and Klarite™ samples produced about a 10-fold increase in signal enhancement compared to gold nanostars for the SERS analysis of aniline, nitroaniline isomers, and nitrobenzene. Signal enhancement is significantly greater for aniline compared to nitrobenzene and it is dependent on the proximity of the NH 2 to the NO 2 group for the nitroaniline isomers. Charge-transfer in chemisorbed analytes is an important contributing factor for SERS signal. The relative strengths of enhancement can be predicted by the DFT calculation of the HOMO-LUMO energy gaps of the analyte-metal cluster. Aniline and the three nitroaniline isomers showed stronger preference for the copper substrates if both the gold and copper substrates are present. The NO 2 group in 2-nitroaniline has a very strong preference and affinity for the copper in the Au-Cu bimetallic cluster.