Alkaline phosphatase (ALP) is integral to a diverse array of biological processes, consequently, its presence is frequently linked to the onset of various pathologies. In the current investigation, 1-naphthyl phosphate (1-Npp) served as a substrate, with the catalytic hydrolysis of phosphate groups by ALP being leveraged to engender a detectable current at an applied potential of 0.277 V. Initially, carbon nanotubes (SWCNTs) were deposited via drop-coating onto graphite paper (Gp), a material characterized by exceptional electrical conductivity. Subsequently, Cu-MOF was uniformly deposited on Gp/SWCNTs composites by rapid electrodeposition technique, that is, chronocurrent method (i–t), at a potential of −1.1 V for 240 s, thereby fabricating an electrochemical sensor platform denominated Gp/SWCNTs/Cu-MOF. This platform enabled the ratiometric electrochemical quantification of ALP, utilizing the signal from Cu-MOF as an internal standard. This platform’s integrated design facilitates signal amplification and provides an inbuilt reference, which was instrumental in mitigating the impact of environmental fluctuations and enhancing the precision of the obtained data. The ratiometric signals exhibited strong linear correlations within the 0.014–14.29 U mL−1 concentration, with a detection limit of 0.005 U mL−1. Moreover, the sensor exhibited robust selectivity, repeatability, and excellent detection performance in real-world samples, with recovery rates spanning 99.3 %–103.5 %, making it a suitable candidate for the expedient and precise quantification of ALP in serum.
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