In situ fluorogenic reaction for ratiometric fluorescent detection of alkaline phosphatase activity

Jialin Wen; Yuxuan Hu; Na Li; Dan Li; Guangfan Zheng; Yongjian Zou; Minmin Zhang; Lingling Shui

doi:10.1016/j.aca.2022.340414

In situ fluorogenic reaction for ratiometric fluorescent detection of alkaline phosphatase activity

Anal Chim Acta. 2022 Oct 16:1230:340414. doi: 10.1016/j.aca.2022.340414. Epub 2022 Sep 19.

Authors

Jialin Wen¹, Yuxuan Hu², Na Li³, Dan Li⁴, Guangfan Zheng⁵, Yongjian Zou², Minmin Zhang², Lingling Shui⁶

Affiliations

¹ Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510006, PR China; South China Academy of Advanced Optoelectronics, South China Normal University, Guangzhou, 510006, PR China.
² Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510006, PR China.
³ Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510006, PR China. Electronic address: dreamerlina@163.com.
⁴ School of Control Engineering, Northeastern University at Qinhuangdao, Qinhuangdao, 066004, PR China. Electronic address: lidan1@neuq.edu.cn.
⁵ School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, 710062, PR China.
⁶ Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510006, PR China; South China Academy of Advanced Optoelectronics, South China Normal University, Guangzhou, 510006, PR China. Electronic address: shuill@m.scnu.edu.cn.

PMID: 36192072
DOI: 10.1016/j.aca.2022.340414

Abstract

Assays utilizing in situ fluorogenic reactions provide a simple and convenient alternative approach for the detection of biological molecules and activities. In this work, a novel ratiometric fluorescent probe based on in situ fluorogenic reaction is explored and developed for alkaline phosphatase (ALP) activity sensing. An intriguing fluorogenic reaction between 2,3-diaminonaphthalene (2,3-DAN) and ascorbic acid (AA) in alkaline aqueous solutions could generate the fluorescent quinoxaline derivative. The resultant quinoxaline emits intense yellow fluorescence, differing from the blue fluorescence of 2,3-DAN. Thus, a ratiometric fluorescent probe based on this fluorogenic reaction is constructed for ALP activity sensing, combining with ALP-triggered hydrolysis of AA2P into AA. Meanwhile, the addition of copper(II) acetate into the reaction system largely improves reaction rate and efficiency. This sensing strategy shows high sensitivity for ALP activity with detection limit of 0.08 U/L, and excellent selectivity towards ALP out of various interferences. This method is extended to human salivary ALP detection. The present method provides a simple and reliable alternative for the detection of ALP activity and has the potential for clinical applications. It also shows a feasible way to construct ratiometric fluorescent methods.

Keywords: 2,3-Diaminonaphthalene; Alkaline phosphatase; Ascorbic acid; In situ fluorogenic reaction; Ratiometric fluorescence.

MeSH terms

Alkaline Phosphatase*
Ascorbic Acid
Fluorescent Dyes*
Humans
Quinoxalines
Spectrometry, Fluorescence / methods

Substances

Fluorescent Dyes
Quinoxalines
Alkaline Phosphatase
Ascorbic Acid