We describe a simple and effective strategy to construct a molecular imprinting ratiometric fluorescence sensor (MIR sensor) for the visual detection of bovine hemoglobin (BHb) used as a model protein. The sensor was prepared by simply mixing the solution of green and red CdTe quantum dots (QDs), which were embedded in core-shell structured molecularly imprinted polymers and silica nanoparticles, respectively. The resultant hybrid MIR sensor can selectively bind with BHb and thus quench the fluorescence of the green QDs, while the red QDs wrapped with silica are insensitive to BHb with the fluorescence intensity unchanged. As a result, a continuous obvious fluorescence color change from green to red can be observed by naked eyes, with the detection limit of 9.6 nM. Moreover, the MIR sensor was successfully applied to determine BHb in bovine urine samples with satisfactory recoveries at three spiking levels ranging from 95.7 to 101.5%, indicating great potential application for detecting BHb in real samples. This strategy of using different fluorescence emission materials incorporated to construct a ratiometric fluorescence sensor is reasonable and convenient, which can be extended to the preparation of other ratiometric fluorescence systems for targeted analytes.

We reported a facile strategy to assemble a ratiometric nanosensor for the ovalbumin (OVA) fluorescence determination and meanwhile it can be utilized for selective visual identification by naked eyes with fluorescent test papers under 365 nm UV lamp. The nanosensor was prepared through simply mixing blue color carbon dots (CDs) and green color core-shell imprinted polymers. Blue CDs were used directly as the internal reference without participating in the imprinting process and modified molecularly imprinted polymers (MIPs) were synthesized by post-imprinting, using fluorescein isothiocyanate (FITC) as fluorescence enhanced signal. Upon the addition of different concentrations of OVA, the fluorescence intensity of FITC was enhanced, while the fluorescence intensity of CDs was almost unchanged, leading to a detection limit as low as 15.4 nM. Accordingly, the fluorescence color was gradually changed from blue to dark olive green to green with naked eyes observation. Moreover, the ratiometric nanosensor was successfully applied to detect OVA in the human urine samples with satisfactory recoveries attaining of 92.0-104.0% with relative standard deviation (RSD) of 3.3-3.9% and 93.3-101.0% with RSDs of 2.7-3.8% for the spiked chicken egg white samples. This strategy reported here opens a novel pathway for biomacromolecule detection in real applications and can realize the visual observation on fluorescent test papers.