In this work, through thegrowth of MnOnanosheets on the surface of terbium metal-organic frameworks (Tb-MOFs), MOF@MnOnanocomposites are prepared and the fluorescence of Tb-MOFs is quenched significantly by MnO. Additionally, the hybrid nanoflowers are self-assembled by cholesterol oxidase (ChOx) and copper phosphate (Cu(PO)·3HO). Then a new strategy for cholesterol determination is developed based on MOF@MnOnanocomposites and hybrid nanoflowers. Cholesterol is oxidized under the catalysis of hybrid nanoflowers to yield HO, which further reduces MnOnanosheets into Mn. Hence, the fluorescence recovery of Tb-MOFs is positively correlated to the concentration of cholesterol in the range of 10 to 360M. The limit of detection (LOD) of cholesterol is 1.57M. On the other hand, the hierarchical and confined structure of ChOx-inorganic hybrid nanoflowers greatly improve the stability of the enzyme. The activity of hybrid nanoflowers remains at a high level for one week when stored at room temperature. Moreover, the hybrid nanoflowers can be collected by centrifugation and reused. The activity of hybrid nanoflowers can continue at a high level for five cycles of determination. Therefore, it can be concluded that the hybrid nanoflowers are more stable and more economic than free enzymes, and they show a similar sensitivity and specificity to cholesterol compared with free ChOx. Finally, this strategy has been further validated for the determination of cholesterol in serum samples with satisfactory recoveries.
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