title: Naked-eye sensitive detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) based on a horseradish peroxidase catalytic colorimetric system with Cu(ii).
abstract: In this paper, a novel colorimetric method for the detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) was designed based on a Cu(2+)-horseradish peroxidase (HRP)-3,3',5,5'-tetra-methylbenzidine (TMB)-H2O2 system. In the presence of ALP, l-ascorbic acid-2-phosphate (AAP) could be hydrolyzed to ascorbic acid which could reduce Cu(2+) 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 Cu(2+) was chelated by PPi, the conversion of Cu(ii) by AA was effectively inhibited. The color of the HRP-TMB-H2O2 system with Cu(2+) showed blue. The HRP-TMB-H2O2 system with the Cu(2+) 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.
authors.affiliation: Key Laboratory of Chem-Biosensing, Anhui province; Key Laboratory of Functional Molecular Solids, Anhui province; College of Chemistry and Materials Science, Center for Nano Science and Technology, Anhui Normal University, Wuhu 241000, PR China. wangyuz@mail.ahnu.edu.cn.
authors.forename: Dongmin
authors.initials: D
authors.lastname: Shi
authors.forename: Yue
authors.initials: Y
authors.lastname: Sun
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.forename: Chunjun
authors.initials: C
authors.lastname: Shi
authors.forename: Guangfeng
authors.initials: G
authors.lastname: Wang
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis*; D003124:Colorimetry / Q000379:methods*; D003300:Copper / Q000737:chemistry*; D011756:Diphosphates / Q000032:analysis*; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D011756:Diphosphates; D003300:Copper; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase