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Use of bovine serum albumin-gold-silver alloy nanocluster for detecting alkaline phosphatase

A technology of bovine serum albumin and gold-silver alloy, which is applied in the field of detection of alkaline phosphatase, can solve the problem that few people care about ALP detection, and achieve the effect of simple, convenient and high sensitivity detection method

Pending Publication Date: 2019-04-09
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is of great significance to develop a fluorescent sensor for alkaline phosphatase. More importantly, the fluorescent detection and analysis method of ALP is basically focused on serum research, but the detection of ALP in the environment is rarely paid attention to.

Method used

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  • Use of bovine serum albumin-gold-silver alloy nanocluster for detecting alkaline phosphatase
  • Use of bovine serum albumin-gold-silver alloy nanocluster for detecting alkaline phosphatase
  • Use of bovine serum albumin-gold-silver alloy nanocluster for detecting alkaline phosphatase

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0027] (1) Preparation of bovine serum albumin-gold-silver alloy nanocluster (BSA-Au / Ag NCs) aqueous solution:

[0028] Mix 4mL of chloroauric acid (10mM) and 1mL of silver nitrate (10mM) (the molar ratio of gold and silver is 4:1), add it to 5mL of bovine serum albumin solution (0.75mM), stir vigorously and mix for 5 minutes , adjust the pH of the solution with sodium hydroxide, and then place it in a constant temperature oil bath at 37°C for 12 hours to react. After the reaction is completed, use ultrapure water for dialysis and purification for 48 hours to obtain a BSA-Au / Ag NCs aqueous solution;

[0029] (2) Add KMnO to the BSA-Au / Ag NCs aqueous solution prepared in step (1) 4 Aqueous solution quenches its fluorescence, makes mixed solution A, and detection fluorescence intensity is I 1 . Among them, BSA-Au / Ag NCs aqueous solution and KMnO 4 The volume ratio of the aqueous solution is 2:1, the concentration of BSA-Au / Ag NCs aqueous solution is 20nmol / L, KMnO 4 The conc...

example 2

[0045] (1) The steps for preparing an aqueous solution of bovine serum albumin-gold-silver alloy nanoclusters (BSA-Au / Ag NCs) are the same as in Example 1.

[0046] (2) mixed solution A preparation step is the same as embodiment 1, and detection fluorescence intensity is 1 1 .

[0047] (3) Alkaline phosphatase catalyzes ascorbic acid 2-phosphate to generate ascorbic acid. The steps are the same as in Example 1.

[0048] (4) The preparation steps of mixed solution B are the same as in Example 1, and the detection fluorescence intensity is 1.

[0049] (5) Establish (I-I 1 ) / I and the standard curve of alkaline phosphatase concentration.

[0050] (6) Detection of alkaline phosphatase in water sample 1:

[0051] Collect lake water sample 1, use a 0.22 μm filter membrane to remove suspended particles in the water sample, dilute 10 times with Tris-HCl buffer, and set aside.

[0052] Add ascorbic acid 2-phosphate (300nmol / L) into the water sample to be tested. After the catalysi...

example 3

[0054] (1) The steps for preparing an aqueous solution of bovine serum albumin-gold-silver alloy nanoclusters (BSA-Au / Ag NCs) are the same as in Example 1.

[0055] (2) mixed solution A preparation step is the same as embodiment 1, and detection fluorescence intensity is 1 1 .

[0056] (3) Alkaline phosphatase catalyzes ascorbic acid 2-phosphate to generate ascorbic acid. The steps are the same as in Example 1.

[0057] (4) The preparation steps of mixed solution B are the same as in Example 1, and the detection fluorescence intensity is 1.

[0058] (5) Establish (I-I 1 ) / I and the standard curve of alkaline phosphatase concentration.

[0059] (6) Detection of alkaline phosphatase in water sample 2:

[0060] Collect lake water sample 1, use a 0.22 μm filter membrane to remove suspended particles in the water sample, dilute 10 times with Tris-HCl buffer, and set aside.

[0061] Add ascorbic acid 2-phosphate (300nmol / L) into the water sample to be tested. After the catalysi...

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Abstract

The invention belongs to the field of analytical chemistry and provides use of a bovine serum albumin-gold-silver alloy nanocluster for detecting alkaline phosphatase, comprising the following steps:1) preparing a BSA-Au / Ag NCs aqueous solution, adding a KMnO4 solution to quench fluorescence of the BSA-Au / Ag NCs aqueous solution to obtain a mixed solution A, and detecting fluorescence intensity to be I1; 2) using alkaline phosphatase to catalyze ascorbic acid 2-phosphate to generate ascorbic acid; 3) mixing the mixed solution A and ascorbic acid to react and detecting the fluorescence intensity I; 4) establishing a standard curve of (I-I1) / I and alkaline phosphatase concentration; and 5) adding the ascorbic acid 2-phosphate to a to-be-tested sample, catalyzing, then, mixing with the mixedsolution A, after the completion of the reaction, detecting the fluorescence intensity I', and comparing the value of (I'-I1) / I' with the standard curve to obtain the alkaline phosphatase concentration in the sample. The method of the invention has the advantages of high sensitivity, good selectivity and simple operation. The alkaline phosphatase level in environmental water is related to the degree of eutrophication of water. The use in the invention has important significance in inventing an alkaline phosphatase fluorescence sensor.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and relates to a use of bovine serum albumin-gold-silver alloy nano-clusters to detect alkaline phosphatase. Background technique [0002] In recent years, water eutrophication has become one of the main causes of water pollution, so it has attracted more and more attention. As we all know, eutrophication of water body is caused by excessive content of nutrients such as nitrogen and phosphorus in water. The influx of large amounts of phosphorus in rivers and lakes leads to eutrophication and algal blooms. Therefore, the monitoring of phosphorus in water is often used to monitor the degree of eutrophication in water. Today, simply measuring the total phosphorus concentration in the water is very useful in discerning the relationship between phosphorus and eutrophication because the directly available inorganic phosphorus content of phytoplankton in lake water is usually less than 5% of the tot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 孙见凡王晓艳刘振江胡文杰杜道林
Owner JIANGSU UNIV
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