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Two-channel detection method of alkaline phosphatase activities

A technology of phosphatase activity and detection method, which is applied in the field of dual-channel detection of alkaline phosphatase activity, can solve the problem of low sensitivity of colorimetric detection and visual semi-quantitative detection, construction of multiple signal sensing system, poor luminescent performance, etc. problems, to achieve the effect of simple preparation process, increased sensitivity and visualization, and high sensitivity

Inactive Publication Date: 2019-01-25
UNIV OF JINAN
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Problems solved by technology

Chinese patent CN104007080A discloses an alkaline phosphatase analysis method based on gold nanorods and long silver shells. Although the wavelength variability of the core-shell nanorods in this method can provide rich color changes, it is difficult to combine them with fluorescence. Combining materials to build multi-signal sensing systems
Conventional undoped graphene quantum dots have poor luminescence performance and low quantum yield, and heteroatom doping can effectively control their optical characteristics and improve the fluorescence quantum yield
Chinese patent CN106769959A discloses a method for detecting alkaline phosphatase activity based on dual modes of fluorescence and colorimetry, which has a long response time, low resolution of colorimetric detection sensitivity and visual semi-quantitative detection

Method used

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  • Two-channel detection method of alkaline phosphatase activities
  • Two-channel detection method of alkaline phosphatase activities
  • Two-channel detection method of alkaline phosphatase activities

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Embodiment 1

[0033] (1) The preparation of GQDs specifically includes the following steps:

[0034] Mix 1g of citric acid and 0.3g of cysteine ​​in a 5mL small beaker and heat it to 240°C with a heating mantle. Colorless to light yellow to orange, forming GQDs. The obtained product was dissolved in ultrapure water and dialyzed with a dialysis bag with a molecular weight cut off of 500 for 24 hours, and the finally obtained GQDs were stored at room temperature in the dark. The particle size of the prepared GQDs is between 1.2-2.6nm, the maximum excitation and emission wavelengths are at 346nm and 415nm respectively, the quantum yield is 67%, and it can be kept stable for at least half a year at room temperature.

[0035] (2) The preparation of AuNPs specifically includes the following steps:

[0036]All glass containers used were soaked in aqua regia (the volume ratio of concentrated hydrochloric acid to concentrated nitric acid was 3:1), washed with ultrapure water and dried for later us...

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Abstract

The invention provides a method for colorimetric-fluorescent two-channel detection of alkaline phosphatase activities. The method comprises the following steps: catalyzing a substrate trisodium L-ascorbyl-2-phosphate (AAP) of alkaline phosphatase by utilizing the alkaline phosphatase so as to hydrolyze to generate ascorbic acid (AA) with reducibility, whereas the AA can cause a Tollens reagent tobe reduced to an Ag elementary substance; under the condition that gold nanoparticles (AuNPs) and graphene quantum dots (GQDs) exist, Ag can be deposited on the surface of AuNPs to form gold and silver core shell nanoparticles (Au@AgNPs) and quenches the fluorescence of GQDs. According to the method, metallization of enzymatic silver ions, AuNPs-induced silver deposition and Au@AgNPs-quenched GQDsfluorescence are combined, the changes of color, ultraviolet-visible absorption spectroscopy and fluorescence intensity are utilized, and the colorimetric-fluorescent two-channel detection of the alkaline phosphatase activities can be realized. The method has the characterics that the sensitivity is high, the specificity is strong, the response is fast, the operation is simple, and no complex instruments are required.

Description

technical field [0001] The invention relates to a dual-channel detection method for alkaline phosphatase activity, belonging to the technical field of enzyme detection. Background technique [0002] Alkaline phosphatase (ALP) is an important biomarker whose abnormal expression is associated with various diseases such as breast cancer, prostate cancer, bone disease, diabetes and liver dysfunction. Quantitative detection of ALP in serum can assist in the diagnosis of related diseases. Spectroscopic analysis has been widely used in ALP detection due to its simplicity and fast response. However, conventional spectroscopic methods based on a single material with a single colorimetric or fluorescent output signal have poor anti-interference ability and low accuracy, making it difficult to detect targets in complex samples. With the advancement of nanotechnology, a variety of ALP colorimetric sensing methods based on nano-gold and nano-silver have been developed, but the sensitiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/64G01N21/31
CPCG01N21/3103G01N21/6402G01N21/6486G01N21/78
Inventor 逯一中陈传霞姜媛媛倪朋娟
Owner UNIV OF JINAN
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