Citation: Mingyue Luo, Kehui Zhang, Honghong Rao, Jianying Li, Xin Xue, Panpan Sun, Xiaoquan Lu, Zhonghua Xue. A simplified ratiometric fluorescent sensing strategy for enhanced detection of alkaline phosphatase employing Prussian blue nanozymes and commercially available chromogen[J]. Chinese Chemical Letters, ;2025, 36(9): 110703. doi: 10.1016/j.cclet.2024.110703 shu

A simplified ratiometric fluorescent sensing strategy for enhanced detection of alkaline phosphatase employing Prussian blue nanozymes and commercially available chromogen

Mingyue Luo ^a ,
Kehui Zhang ^b ,
Honghong Rao ^b ,
Jianying Li ^a ,
Xin Xue ^c ,
Panpan Sun ^a ,
Xiaoquan Lu ^a ,
Zhonghua Xue ^{a, *,}

a.
Key Laboratory of Water Security and Water Environment Protection in Plateau Intersection (NWNU), Ministry of Education, Key Lab of Bioelectrochemistry and Environmental Analysis of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070, China
b.
School of Chemical Engineering, Lanzhou City University, Lanzhou 730070, China
c.
State Key Laboratory of Applied Organic Chemistry and Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, Lanzhou University, Lanzhou 730000, China

xzhlab@hotmail.com

xzh@nwnu.edu.cn

Received Date: 27 September 2024
Revised Date: 22 November 2024
Accepted Date: 2 December 2024
Available Online: 3 December 2024

Figures(6)

The traditional nanozymes-based ratiometric fluorescence sensing platforms usually necessitate the supplementary addition of fluorescent probes, therefore greatly restricting its convenient and broad application. In this study, a highly sensitive and selective ratiometric fluorescence platform for alkaline phosphatase (ALP) detection was established, only employing Prussian blue (PB) nanozymes and a commercially available chromogen of o-phenylenediamine (OPD). PB nanozymes with remarkable peroxidase-like (POD-like) activity can effectively catalyze OPD chromogen to yield 2,3-diaminophenazine (OPDox) with an intense yellow fluorescence at 573 nm emission peak. Target ALP can facilitate ascorbic acid 2-phosphate (AAP) dephosphorylation to generate phosphate and ascorbic acid (AA). Significantly, both these two resultant hydrolysis products could effectively decrease the OPDox generation via a dual-path based inhibition on the PB nanozymes POD-like activity. On the other hand, the generated dehydroascorbic acid (DHAA) from AA oxidation would exclusively react with OPD chromogen to yield 3-(dihydroxyethyl)furo[3,4-b]quinoxaline-1-one (DFQ) with a strong blue fluorescent signal at 434 nm, which further providing a significant enhancement on the sensing selectivity of ALP detection. As a result, an increased yellow fluorescence of OPDox and decreased blue fluorescence of DFQ could be clearly observed with different ALP addition. A robust linear relationship between the fluorescence ratio of F₄₃₄/F₅₇₃ and ALP activity ranging from 0.25 U/L to 6 U/L was obtained, with a low detection limit of 0.112 U/L. This proposed method demonstrates high sensitivity, excellent selectivity, cost-effectiveness, and operational simplicity, yet enabling an effective detection of ALP levels in human serum.
- Ratiometric fluorescence,
- Prussian blue nanozymes,
- Peroxidase-like activity,
- Commercially available chromogens,
- Alkaline phosphatase
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