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Method for detecting chiral compound based on aptamer modified nanogold

A technology for chiral compounds and nucleic acid aptamers, which is used in the fields of chiral identification, colorimetric analysis and environmental detection, and can solve the problems of long analysis time, unfavorable rapid identification, expensive chiral chromatographic columns, and high separation conditions. , to reduce the detection cost and the complexity of the detection scheme, the modification is convenient and fast, and the effect of non-immunogenicity

Inactive Publication Date: 2015-09-09
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The simplest is the optical rotation method, but the optical rotation direction of the same optically active compound will change due to the change of the measurement conditions, and there is no easy-to-distinguish relationship with the three-dimensional structure. It is easy to identify chiral compounds by measuring the optical rotation. limited
High-performance liquid chromatography, capillary electrophoresis, etc. are realized by combining separation means with photometric detection, but the chiral chromatographic columns often required are expensive, consume a lot of reagents and have high requirements for separation conditions, and the analysis time is not long enough. for quick identification
However, spectroscopic analysis and electrochemical sensors have low sensitivity and complicated identification process.

Method used

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  • Method for detecting chiral compound based on aptamer modified nanogold
  • Method for detecting chiral compound based on aptamer modified nanogold
  • Method for detecting chiral compound based on aptamer modified nanogold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Qualitative and quantitative chiral recognition of S-ibuprofen in the environment with S-ibuprofen aptamer-probe complex

[0029] (1): Preparation of gold nanoparticle solution:

[0030] Add 50mL of ultrapure water into the 100mL round bottom beaker soaked and cleaned by aqua regia, pipette 0.5mL of 1% chloroauric acid aqueous solution prepared in advance into 50mL of ultrapure water with a cleaned 1mL pipette, The concentration of chloroauric acid in the solution was reduced to 0.01% (w / v), and the solution was heated in an oil bath at a constant temperature of 92±4°C. Under the vigorous stirring of the magnet, quickly add 1.6mL of 1% (w / v) trisodium citrate solution prepared in advance, and continue to stir. In the first three minutes of the reaction, the solution basically does not change. After 3 minutes, the solution begins to Turn blue, turn into blue-purple after a few minutes, continue to stir at constant temperature, the solution turns red after 20 ...

Embodiment 2

[0039] Example 2: Qualitative and quantitative chiral recognition of R-ibuprofen in the environment with R-ibuprofen aptamer-probe complex

[0040] (1): Preparation of gold nanoparticle solution: the specific steps are the same as in Example 1.

[0041] (2): prepare the R-ibuprofen aptamer, the sequence of the R-ibuprofen aptamer is:

[0042] 5'-SH GCGAACGACTTCATAAAATGCTATAAGGTTGCCCTCTGTC-3'. After centrifuging the obtained R-ibuprofen aptamer, add Tris-HCl buffer solution to dissolve into a 1 μmol / L solution, and put it in a 4°C refrigerator for later use;

[0043] (3): Using the R-Ibuprofen aptamer as the recognition probe for R-Ibuprofen, add 200 μL of the above-mentioned nano-gold solution and 100 μL of the above-mentioned R-Ibuprofen aptamer solution in a 2 mL centrifuge tube, Add 0, 20, 40, 60, 80, 100, and 120 μL of 100 μmol R-ibuprofen standard solution to 10 μL of 1M sodium chloride solution, and dilute to 500 μL with ultrapure water to obtain a standard detection s...

Embodiment 3

[0049] Example 3: Qualitative and quantitative chiral recognition of S and R-Ibuprofen mixtures with S-Ibuprofen aptamer-probe complex

[0050] (1): Preparation of gold nanoparticle solution: the specific steps are the same as in Example 1.

[0051] (2): Prepare the S-ibuprofen aptamer, the specific steps are the same as in Example 1.

[0052] (3) Using the S-Ibuprofen aptamer as the recognition probe for S-Ibuprofen, add 200 μL of the above-mentioned gold nanoparticle solution and 100 μL of the above-mentioned S-Ibuprofen aptamer solution in a 2 mL centrifuge tube, To 10 μL of 1M sodium chloride solution, add 0, 20, 40, 60, 80, 100, and 120 μL of S-ibuprofen solution with a concentration of 100 μmol and correspondingly add 120, 100, 80, 60, 40, 20, and 0 μL of 100 μmol of R-ibuprofen solution was mixed, and the volume was adjusted to 500 μL with ultrapure water. After reacting for 30 minutes at room temperature, scan with UV-vis in the wavelength range of 190-800nm. Accordi...

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Abstract

The invention provides a method for detecting chiral compound based on aptamer modified nanogold. The method is characterized by specifically comprising the following steps: step 1, preparing a gold nanoparticle solution; step 2, preparing at least one of an S-ibuprofen aptamer and an R-ibuprofen aptamer; and step 3, by taking the S-ibuprofen aptamer as a recognition probe of S-ibuprofen, mixing the gold nanoparticle solution, S-ibuprofen aptamer solution, sodium chloride solution and S-ibuprofen standard solution, making the volume constant, reacting for 30 minutes, detecting the light absorption values A650 and A520 of the detection system at 650nm and 520nm by virtue of an ultraviolet spectrophotometer, and obtaining a standard curve by taking the ibuprofen concentration as a horizontal coordinate and taking the A650 / A520 as a vertical coordinate; and detecting the S-ibuprofen concentration in a to-be-detected sample by utilizing the standard curve. The method disclosed by the invention can be finished in a short time, qualitative and quantitative detection results are obtained, the method has the advantages of high specificity, high stability, wide application range, low cost and the like, and the conventional application of chiral recognition is easily popularized.

Description

technical field [0001] The invention relates to the technical fields of chiral recognition, colorimetric analysis and environmental detection, in particular to an aptamer colorimetric detection method for detecting S-ibuprofen and R-ibuprofen. Background technique [0002] The existence of chiral enantiomers is a common phenomenon in nature. Biomacromolecules, such as proteins, polysaccharides, nucleic acids, and enzymes, are almost all chiral, and these small molecules often have important functions in the body. Physiological function. During the process of absorption, distribution, metabolism and excretion, chiral enantiomers produce different pharmacological effects and reactions through different three-dimensional combinations with macromolecules in the body. In terms of clinical treatment, taking enantiomerically pure chiral drugs can not only eliminate the toxic and side effects caused by ineffective (bad) enantiomers, but also reduce the dosage and the metabolic burd...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33G01N21/78
Inventor 平婧柳建设沈忱思刘娜娄晓祎陈名明于海波
Owner DONGHUA UNIV
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