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Herein we demonstrate the synthesis, characterization, and peroxidase-mimicking behavior of carboxymethyl cellulose-Co3O4nanoparticles (C@NZ3) toward rapid and sensitive colorimetric detection of ascorbic acid (AA). AA was detected by monitoring the quenching of color signal that was obtained from C@NZ3catalyzed H2O2oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB). The excellent peroxidase-mimicking behavior of C@NZ3is evidenced by the low detection limit of AA (0.41 μM) and good linearity in the range of 0.5–30 μM. The method is cost-effective, highly selective, and requires only 3 min to complete. C@NZ3exhibited high shelf life and could be successfully used to detect AA in real samples of human serum.