Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction

Development of sensitive and convenient alkaline phosphatase (ALP) detection methods is of great significance for food analysis, biomedical applications, and clinical tests. In this work, a sensitive detection method for ALP was established based on nanochannel-modified electrodes, where the electro...

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Main Authors: Tianjun Ma, Xuan Luo, Fengna Xi, Nuo Yang
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Biosensors
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Online Access:https://www.mdpi.com/2079-6374/15/6/377
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author Tianjun Ma
Xuan Luo
Fengna Xi
Nuo Yang
author_facet Tianjun Ma
Xuan Luo
Fengna Xi
Nuo Yang
author_sort Tianjun Ma
collection DOAJ
description Development of sensitive and convenient alkaline phosphatase (ALP) detection methods is of great significance for food analysis, biomedical applications, and clinical tests. In this work, a sensitive detection method for ALP was established based on nanochannel-modified electrodes, where the electrochemical luminescence (ECL) signal was quenched by the enzymatic reaction product. Vertically ordered mesoporous silica film (VMSF) was rapidly grown on low-cost ITO via the electrochemically assisted self-assembly (EASA) method. The resulting modified electrode (VMSF/ITO) exhibited a uniform and ordered nanochannel structure with nanochannel diameter of 2–3 nm and charge-selective permeability, enabling the enrichment of cationic ECL emitter tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)<sub>3</sub><sup>2+</sup>). Compared to the ITO electrode, VMSF/ITO increased the ECL signal by 60 times. In the presence of ALP, it catalyzes the hydrolysis of its substrate, disodium phenyl phosphate hydrate (DPP), generating phenol (Phe), which quenched the ECL signal of Ru(bpy)<sub>3</sub><sup>2</sup><sup>+</sup> and the co-reactant N,N-Dipropyl-1-propanamine (TPA). Based on this principle, ECL detection of ALP can be achieved. The linear detection range for ALP was 0.01 U/L to 30 U/L, with a limit of detection (LOD) of 0.008 U/L. The sensor exhibited high selectivity. Combined with the anti-contamination and anti-interference capabilities of VMSF, the constructed sensor enabled the detection of ALP levels in milk samples.
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spelling doaj-art-7b1a461b436c45f48429b7fbc667fdc72025-08-20T03:27:22ZengMDPI AGBiosensors2079-63742025-06-0115637710.3390/bios15060377Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic ReactionTianjun Ma0Xuan Luo1Fengna Xi2Nuo Yang3The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, ChinaSchool of Chemistry and Chemical Engineering, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaSchool of Chemistry and Chemical Engineering, Zhejiang Sci-Tech University, Hangzhou 310018, ChinaThe First Affiliated Hospital of Guangxi Medical University, Nanning 530021, ChinaDevelopment of sensitive and convenient alkaline phosphatase (ALP) detection methods is of great significance for food analysis, biomedical applications, and clinical tests. In this work, a sensitive detection method for ALP was established based on nanochannel-modified electrodes, where the electrochemical luminescence (ECL) signal was quenched by the enzymatic reaction product. Vertically ordered mesoporous silica film (VMSF) was rapidly grown on low-cost ITO via the electrochemically assisted self-assembly (EASA) method. The resulting modified electrode (VMSF/ITO) exhibited a uniform and ordered nanochannel structure with nanochannel diameter of 2–3 nm and charge-selective permeability, enabling the enrichment of cationic ECL emitter tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)<sub>3</sub><sup>2+</sup>). Compared to the ITO electrode, VMSF/ITO increased the ECL signal by 60 times. In the presence of ALP, it catalyzes the hydrolysis of its substrate, disodium phenyl phosphate hydrate (DPP), generating phenol (Phe), which quenched the ECL signal of Ru(bpy)<sub>3</sub><sup>2</sup><sup>+</sup> and the co-reactant N,N-Dipropyl-1-propanamine (TPA). Based on this principle, ECL detection of ALP can be achieved. The linear detection range for ALP was 0.01 U/L to 30 U/L, with a limit of detection (LOD) of 0.008 U/L. The sensor exhibited high selectivity. Combined with the anti-contamination and anti-interference capabilities of VMSF, the constructed sensor enabled the detection of ALP levels in milk samples.https://www.mdpi.com/2079-6374/15/6/377alkaline phosphataseelectrochemiluminescencehomogeneous sensingnanochannel-modified electrodesignal quenching
spellingShingle Tianjun Ma
Xuan Luo
Fengna Xi
Nuo Yang
Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction
Biosensors
alkaline phosphatase
electrochemiluminescence
homogeneous sensing
nanochannel-modified electrode
signal quenching
title Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction
title_full Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction
title_fullStr Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction
title_full_unstemmed Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction
title_short Simple Nanochannel-Modified Electrode for Sensitive Detection of Alkaline Phosphatase Through Electrochemiluminescence Signal Quenching by Enzymatic Reaction
title_sort simple nanochannel modified electrode for sensitive detection of alkaline phosphatase through electrochemiluminescence signal quenching by enzymatic reaction
topic alkaline phosphatase
electrochemiluminescence
homogeneous sensing
nanochannel-modified electrode
signal quenching
url https://www.mdpi.com/2079-6374/15/6/377
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AT fengnaxi simplenanochannelmodifiedelectrodeforsensitivedetectionofalkalinephosphatasethroughelectrochemiluminescencesignalquenchingbyenzymaticreaction
AT nuoyang simplenanochannelmodifiedelectrodeforsensitivedetectionofalkalinephosphatasethroughelectrochemiluminescencesignalquenchingbyenzymaticreaction