Oxytetracycline SERS detection method based on nanomaterial self-assembly

A technology of nanomaterials and detection methods, applied in the field of nanomaterials and analytical chemistry, can solve the problems of drug-resistant strains, human health damage, human calcium deficiency, etc., and achieve the effects of easy chemical modification, good stability and strong specificity

Active Publication Date: 2016-10-26
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But at the same time, oxytetracycline can combine with calcium in the human body, leading to calcium defic

Method used

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  • Oxytetracycline SERS detection method based on nanomaterial self-assembly
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  • Oxytetracycline SERS detection method based on nanomaterial self-assembly

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Embodiment 1

[0031] Through the two-step growth method, the preparation of gold nanoparticles of about 80nm includes the following steps:

[0032] 1) At first, synthesize the gold nanoparticle of about 15nm, by the following steps: get 2.5mL chloroauric acid solution (0.2%) and join in 50mL water and boil, add 2mL sodium citrate (1%, containing 0.05% citric acid ), the solution was boiled for 5min, cooled naturally, and carried out the characterization of ultraviolet and transmission electron microscopy ( figure 2 A. figure 2 B).

[0033] 2) Next, carry out the first step of growth, the specific steps are as follows: Dilute 3mL of gold nanoparticles (about 15nm) to 20mL and add them to the round bottom flask, and add 10mL of gold chloride to the round bottom flask in turn under vigorous stirring. A mixture of acid solution (0.04%) and 10 mL of ascorbic acid (1%) and sodium citrate (1%) was stirred vigorously for 45 minutes, then heated and boiled for 30 minutes, and cooled naturally to...

Embodiment 2

[0036] The activation of the oxytetracycline aptamer, the specific steps are as follows:

[0037] First, the aqueous solution of the aptamer was annealed in a water bath at 65°C for 30 min, and after it was slowly and naturally cooled to room temperature, 1% 1 mM tris(2-carboxyethyl)phosphine hydrochloride was added to the aptamer volume. salt at room temperature for 1 h to activate the sulfhydryl-modified stem-loop DNA.

Embodiment 3

[0039] The connection between colloidal gold and oxytetracycline aptamer, the steps are as follows:

[0040] Take 1mL of 80nm colloidal gold and add the same volume of 1μM oxytetracycline aptamer solution to it, and incubate on a shaker at 37°C for 12h. After incubation, use buffer (NaCl 5mM, Tris 5mM) to centrifuge at 3500rpm for 15min, wash 3 times, collect supernatant and material respectively, and carry out ultraviolet characterization ( figure 2 A) and the determination of nucleic acid content ( figure 2 B) After washing, use buffer to make up the reaction system to the original volume.

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Abstract

The invention provides an oxytetracycline SERS detection method based on nanomaterial self-assembly. The oxytetracycline SERS detection method is used for detecting content of oxytetracycline (OTC) in aquatic products such as fish meal. According to the OTC SERS detection method, gold nanoparticles (80 nm particle size) are connected with gold nanoparticles (15 nm particle size) through a base sequence containing OTC aptamers, and 4-mercaptobenzoic acid is incubated on the surfaces of the gold nanoparticles, thereby generating Raman signals. When the OTC is in contact with a detection system, the OTC aptamers are in specific binding with the OTC, thus the distance between the 80nm gold nanoparticles and the 15nm gold nanoparticles is reduced, the hotspot is enhanced, and the Raman signals are enhanced, thereby achieving the detection purpose. The detection system can detect the oxytetracycline quantitatively, the linearity range is from 0.046 to 460 fg/mL, and the detection limit is 0.004 fg/mL. The oxytetracycline SERS detection method used for oxytetracycline detection has the advantages of being high in sensitivity and being fast and convenient, can be applied to detection of samples such as fish meal, and is accurate and reliable in result.

Description

technical field [0001] The invention relates to the technical field of nanomaterials and analytical chemistry, in particular to an oxytetracycline SERS detection method based on self-assembly of nanomaterials, which is used for detecting oxytetracycline in food. Background technique [0002] Nanomaterial refers to any type of material composed of nanostructure units, and its particle size is generally 0.1-100nm. Nanomaterials have different physical and chemical properties from macroscopic materials, including surface effects, small size effects, quantum size effects, macroscopic quantum tunneling effects, catalytic properties, etc. These characteristics make them have excellent innate application advantages in some aspects. [0003] Nucleic acid aptamer (Aptamer) is obtained from in vitro screening by systematic evolution of ligands by exponential enrichment (SELEX) technology. nucleotide sequence. The high-level structure formed by this oligonucleotide sequence can speci...

Claims

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Application Information

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IPC IPC(8): G01J3/44G01N1/28G01N1/44
CPCG01J3/44G01N1/28G01N1/44
Inventor 王周平孟凡伟马小媛段诺吴世嘉
Owner JIANGNAN UNIV
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