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The NiMn2O4/C necklace-like microspheres (NLM) were successfully prepared by hydrothermal method and oil bath. This unique necklace-like structure makes them exhibit the enhanced intrinsic oxidase-like activity, as the special interface can help capture electrons from 3,3',5,5'-tetramethylbenzidine. The fabricated NiMn2O4/C NLM were successfully used as the high-performance oxidase mimetic to catalyze the oxidation of TMB directly for the color reaction. A simple colorimetric method for detection of ascorbic acid by fading was developed