Journal
MICROCHIMICA ACTA
Volume 184, Issue 2, Pages 541-546Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-016-2033-y
Keywords
Visual assay; Uricase; Peroxidase; Enzyme inhibition; Hydrogen peroxide; AFS; Water analysis; Tetramethylbenzidine
Categories
Funding
- National Natural Science Foundation of China [21475013, 21305009]
- China Postdoctoral Science Foundation [2015 M570773, 2016 T90840]
- scientific research and innovation team in University of Sichuan Provincial Department of Education [15TD0009]
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The article describes a bienzyme visual system for aptamer-based assay of Hg(II) at nanomolar levels. The detection scheme is based on the finding that Hg(II) ions captured by aptamer-functionalized magnetic beads are capable of inhibiting the enzymatic activity of uricase and thus affect the formation of H2O2 and the blue product, i.e., oxidized tetramethylbenzidine. This strategy allows for a visual detection of Hg(II) at nanomolar levels without additional amplification procedure. Measuring the absorbance at 650 nm, the logarithmic calibration plot is linear in the concentration range of 0.5-50 nM and the limit of detection (LOD) is 0.15 nM. This is as low as the LOD obtained by atomic fluorescence spectrometry (AFS). The ions K+, Mg2+, Na+, Ca2+, Cu2+, Zn2+, Fe3+, Al3+, Co2+, AsO2 (-), Ni2+, Cd2+ and Pb2+ do not have a significant effect on color formation. The method was applied to the analysis of (spiked) river water, lake water, mineral water, tap water and certified reference water samples, and the results agreed well with those obtained by AFS or certified values, with recoveries ranging from 97% to 109%. The relative standard deviation for five parallel detections at a 10 nM Hg(II) level is 5.2%.
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