Intrinsic peroxidase-like activity of mesoporous nickel oxide for selective cysteine sensing

Mesoporous nickel oxide nanoflowers (NiO NFs) can be easily synthesized by a two-step synthetic procedure based on modified hydrothermal (MHT) treatment of nickel acetate and ethanol amine in water followed by thermal decomposition at 350 °C for 4 h. After thermal treatment, the porosity is increased by 18% with retention of parental nickel hydroxide size. In this study, for the first time, a new catalytic application of NiO NFs has been revealed in terms of peroxidase-like activity where colorless 3,3',5,5' tetramethylbenzidine (TMB) is oxidized to blue color product in the presence of H₂O₂ at room temperature. Comparative study confirms that mesoporous NiO NFs exhibit superior catalytic activity to the parent analogues, i.e. Ni(OH)₂ or bulk NiO. This intrinsic peroxidase-like activity from an easily synthesized inorganic nanomaterial provides an alternative to horseradish peroxidase (HRP) enzyme. The lower Michaelis constant (K_m) value indicates that the catalyst NiO NFs bind efficiently to the test substrate, i.e. TMB. Interestingly, the NiO NFs-catalyzed TMB oxidation, i.e. blue color formation, has been found to be selectively and successively inhibited by a variable amount of cysteine among a set of 21 congeners. Thus our adopted simple, low-cost and novel colorimetric assay stands to be a highly efficient approach for selective detection of cysteine with a limit of detection (LOD) value of ∼1.1 μM using a simple UV-vis spectrophotometer. The proposed method also exhibits outstanding selectivity and accuracy for N-acetyl cysteine (an analogue of cysteine) estimation in real pharmaceutical samples.