The authors report that cobalt oxyhydroxide (CoOOH) nanoflakes possess intrinsic oxidizing ability to directly oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to form a blue colored product (oxTMB) even in the absence of H2O2 and oxygen. In the presence of ascorbic acid (AA), less of the blue product is formed because AA reduces oxTMB. These findings constitute a new scheme for colorimetric detection of AA. Absorbance, best measured at 652 nm, linearly drops in the 10 nM to 1 μM AA concentration range, and the limit of detection is 5 nM (at an S/N ratio of 3). The reaction is complete within <5 s and highly selective. A strip test has been designed for fast and on-spot visual detection of AA. The method was applied to the direct estimation of AA in the microdialysate of brain, and also in soft drink samples. The strip test is considered to be a promising tool for the rapid screening of AA in brain and commercial samples. Graphic abstract Schematic of the CoOOH-TMB colorimetric system that exhibits a high selectivity for ascorbic acid (AA). A strip test has been designed for fast and on-spot visual detection of AA.