Journal
ANALYST
Volume 141, Issue 19, Pages 5549-5554Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c6an01124a
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Funding
- National Natural Science Foundation of China [21371007]
- Anhui Provincial Natural Science Foundation [1208085QB28]
- Anhui Provincial Natural Science Foundation for Distinguished Youth [1408085J03]
- Natural Science Foundation of Anhui [KJ2012A139]
- Program for Innovative Research Team at Anhui Normal University
- Open funding of State Key Laboratory of Chemo/Biosensing and Chemometrics
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In this paper, a novel colorimetric method for the detection of alkaline phosphatase ( ALP) and pyrophosphate (PPi) was designed based on a Cu2+-horseradish peroxidase (HRP)-3,3', 5,5'-tetra-methyl-benzidine (TMB)-H2O2 system. In the presence of ALP, L-ascorbic acid-2-phosphate (AAP) could be hydrolyzed to ascorbic acid which could reduce Cu2+ to Cu+ to inhibit the enzymatic activity of HRP in the colorimetric system. The change in absorbance was found to be proportional to the ALP concentration with a linear detection range and a limit of detection of 5.4 mU mL(-1). In the presence of PPi, because Cu2+ was chelated by PPi, the conversion of Cu(II) by AA was effectively inhibited. The color of the HRP-TMB-H2O2 system with Cu2+ showed blue. The HRP-TMB-H2O2 system with the Cu2+ colorimetric system could also detect PPi with a satisfying result. In summary, this method possesses sensitivity, reproducibility, and cost-effectiveness without labelling and separation and the use of a colorimetric method is more in line with the requirements of on-site detection and green chemistry.
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