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Fluorometric and colorimetric dual-readout alkaline phosphatase activity assay based on enzymatically induced formation of colored Au@Ag nanoparticles and an inner filter effect. 2019

Chuanxia Chen, and Guanglu Zhang, and Pengjuan Ni, and Yuanyuan Jiang, and Yizhong Lu, and Zhengliang Lu
School of Materials Science and Engineering, University of Jinan, Jinan, 250022, China. mse_chencx@ujn.edu.cn.

An ultrasensitive fluorometric and colorimetric dual-mode assay is described for the determination of the activity of alkaline phosphatase (ALP). ALP catalyzes the decomposition of 2-phospho-L-ascorbic acid, and the ascorbic acid thus generated reduces silver ions. In the presence of gold nanoparticles, gold-silver nanoparticles (Au@Ag NPs) are formed. This is accompanied by a color change form pink to deep yellow. The Au@Ag NPs reduce the fluorescence of blue fluorescent graphene quantum dots due to spectral overlap. The changes of absorbance (measured at 410 and 520 nm) and fluorescence (measured at excitation/emission wavelengths of 346/415 nm) correlate well with the ALP activity in the 0.01-6 mU·mL-1 (absorption) and 0.01-2 mU·mL-1 (fluorescence) ranges, and the detection limits are 9 and 5 μU·mL-1 individually. Graphical abstract Schematic presentation of colorimetric and fluorometric dual-readout assay for alkaline phosphatase (ALP) activity. It is based on enzymatically induced formation of gold-silver nanoparticles (Au@Ag NPs), and the fluorescence quenching of graphene quantum dots due to inner filter effect.

UI MeSH Term Description Entries
D010084 Oxidation-Reduction A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471). Redox,Oxidation Reduction
D003124 Colorimetry Any technique by which an unknown color is evaluated in terms of standard colors. The technique may be visual, photoelectric, or indirect by means of spectrophotometry. It is used in chemistry and physics. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
D006046 Gold A yellow metallic element with the atomic symbol Au, atomic number 79, and atomic weight 197. It is used in jewelry, goldplating of other metals, as currency, and in dental restoration. Many of its clinical applications, such as ANTIRHEUMATIC AGENTS, are in the form of its salts.
D006108 Graphite An allotropic form of carbon that is used in pencils, as a lubricant, and in matches and explosives. It is obtained by mining and its dust can cause lung irritation. Graphene
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000328 Adult A person having attained full growth or maturity. Adults are of 19 through 44 years of age. For a person between 19 and 24 years of age, YOUNG ADULT is available. Adults
D000469 Alkaline Phosphatase An enzyme that catalyzes the conversion of an orthophosphoric monoester and water to an alcohol and orthophosphate. EC 3.1.3.1.
D001205 Ascorbic Acid A six carbon compound related to glucose. It is found naturally in citrus fruits and many vegetables. Ascorbic acid is an essential nutrient in human diets, and necessary to maintain connective tissue and bone. Its biologically active form, vitamin C, functions as a reducing agent and coenzyme in several metabolic pathways. Vitamin C is considered an antioxidant. Vitamin C,Ascorbic Acid, Monosodium Salt,Ferrous Ascorbate,Hybrin,L-Ascorbic Acid,Magnesium Ascorbate,Magnesium Ascorbicum,Magnesium di-L-Ascorbate,Magnorbin,Sodium Ascorbate,Acid, Ascorbic,Acid, L-Ascorbic,Ascorbate, Ferrous,Ascorbate, Magnesium,Ascorbate, Sodium,L Ascorbic Acid,Magnesium di L Ascorbate,di-L-Ascorbate, Magnesium
D012834 Silver An element with the atomic symbol Ag, atomic number 47, and atomic weight 107.87. It is a soft metal that is used medically in surgical instruments, dental prostheses, and alloys. Long-continued use of silver salts can lead to a form of poisoning known as ARGYRIA.
D013050 Spectrometry, Fluorescence Measurement of the intensity and quality of fluorescence. Fluorescence Spectrophotometry,Fluorescence Spectroscopy,Spectrofluorometry,Fluorescence Spectrometry,Spectrophotometry, Fluorescence,Spectroscopy, Fluorescence

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