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With amino acids as model systems, optically active sum frequency generation (OA-SFG) was used to probe the chirality of molecules with a chiral center and an intrinsically achiral chromophore in isotropic solution for the first time. Like that of circular dichroism (CD), the OA-SFG’s near electronic resonance originates from the extrachromophoric chiral perturbation on the carboxyl chromophore. The difference in the relative strengths of OA-SFG and CD among different amino acids can be explained by the difference in the details of perturbations.