Preparation and application of aptamer-magnetic nanoparticle used for enrichment and separation of aflatoxin B1

A technology of magnetic nanoparticles and aflatoxin, which is applied in the field of purification and treatment process of aflatoxin B1 samples, can solve the problems of low separation efficiency, complicated purification and separation operations, etc., and achieves the effect of strong selectivity and high efficiency

Inactive Publication Date: 2015-10-07
INST OF AGRI PROD QUALITY SAFETY & STANDARD JIANGXI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the above-mentioned technical problems, the object of the present invention is to provide an aptamer-magnetic nanoparticle complex for the separation of aflatoxin B1, and further, the present invention also provides an adapter for the separation of aflatoxin B1 The preparation of body-magnetic nanoparticle complex, further, the present invention also relates to the application of a kind of aptamer-magnetic nanocomposite that is used for the separation of aflatoxin B1, to solve the sample purification of aflatoxin B1 in background technology Complicated separation operation and low separation efficiency

Method used

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  • Preparation and application of aptamer-magnetic nanoparticle used for enrichment and separation of aflatoxin B1
  • Preparation and application of aptamer-magnetic nanoparticle used for enrichment and separation of aflatoxin B1
  • Preparation and application of aptamer-magnetic nanoparticle used for enrichment and separation of aflatoxin B1

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The method for preparing the aflatoxin B1 aptamer-magnetic nanoparticle complex comprises the following steps:

[0037] (1) Preparation method of inner core layer of magnetic nanoparticles

[0038] FeCl 3 ·6H 2 O and FeCl 2 4H 2 O was dissolved in 160 mL of 0.1 mol / L hydrochloric acid solution at a molar ratio of 2:1. Transfer the mixed solution into a 250mL flask, stir vigorously for 30min, slowly add 5mol / L NaOH solution dropwise at 40°C, adjust the pH to 10-11, raise the temperature to 60°C, and age for 2 hours. The whole process is carried out under the protection of nitrogen. The magnetic nanoparticles were washed to neutral with deionized water, and the magnetic nanoparticles were freeze-dried.

[0039] (2) Preparation method of magnetic nanoparticle silanization layer

[0040] Take by weighing 200mg of Fe after step (1) freeze-drying 3 o 4 Add 5-15 mL of toluene solution containing 5% 3-(2,3-glycidoxy)propyltrimethoxysilane (volume fraction) to the nanopa...

Embodiment 2

[0046] The aflatoxin B1 aptamer-magnetic nanoparticle extraction method for aflatoxin B1 in a wheat sample comprises the following steps:

[0047] Add 1.0 μg / kg, 2.0 μg / kg, and 5.0 μg / kg aflatoxin B1 to the blank wheat flour, add 5 in parallel, and perform HPLC-FLD analysis after the following steps (1) to (3). The obtained results are shown in Table 1, and the measured recovery rate all reaches more than 80%, and the relative standard deviation is less than 10%, indicating that this nanomaterial is suitable for the purification of aflatoxin B1 in the actual sample of wheat flour;

[0048] (1) Weigh about 50% wheat flour, add 10% water / acetonitrile solution (4:6, v / v), vortex mix for 2min, ultrasonicate for 15min, take 1mL supernatant after standing, and dilute to 10mL with PBS buffer.

[0049] (2) Add the aptamer-magnetic nanoparticle complex containing 10 mg into the solution in step (1), and shake at 37° C. for 30 min. After the reaction, the supernatant was discarded unde...

Embodiment 3

[0056] Establishment of HPLC-FLD evaluation method for aflatoxin B1:

[0057] Liquid phase conditions: detection conditions are:

[0058] Chromatographic column: C18, 4.6×150mm, 5μm, or equivalent;

[0059] Column temperature: 30°C;

[0060] Mobile phase: acetonitrile + methanol + water (13+12+75);

[0061] Flow rate: 1.0mL / min;

[0062] Injection volume: 20.0μL;

[0063] Elution method: isocratic elution;

[0064] Fluorescence detection conditions: excitation wavelength, 365nm, emission wavelength, 440nm.

[0065] Take 50.0μg / L aflatoxin B1 standard solution to make series concentration (1.0, 2.0, 5.0, 10.0, 20.0μg / L), add 20.0μL, HLPC-FLD analysis. Fit it linearly with concentration, the linear equation is Y=1.16×10 6 -2.91×10 6 (X, μg / L; R=0.9906).

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Abstract

The invention relates to preparation and application of aptamer-magnetic nanoparticle used for separation of aflatoxin B1. The compound is composed of the following three layers: (1) an inner nuclear layer composed of paramagnetic Fe3O4 magnetic nanoparticles; (2) a silanization layer coating the inner nuclear layer and having an epoxy function group with reaction activity at the tail end; and (3) a modification layer coupled with aflatoxin B1 aptamer and located on the surface of a shell layer. The invention further provides a method for effectively and quickly separating a trace amount of aflatoxin B1 in a sample by using the compound.

Description

technical field [0001] The invention relates to a process for purifying and treating aflatoxin B1 samples, in particular to a preparation and application method of an aptamer-magnetic nanoparticle separated from aflatoxin B1. Background technique [0002] Aflatoxin B1 is a metabolite produced by Aspergillus flavus and Aspergillus parasiticus. It has carcinogenic, teratogenic and mutagenic effects. It is the most stable mycotoxin found so far. It is not easy to be destroyed by general food processing conditions, so It has laid a huge hidden danger to the food safety of consumers. Countries attach great importance to the status quo of aflatoxin residues in food and have formulated corresponding limit standards. For example, my country stipulates that the allowable amount of aflatoxin in rice and edible oil is (B1+B2+G1+G2) 10ng / g. [0003] At present, the pretreatment technologies for aflatoxin B1 include immunoaffinity columns, multifunctional purification columns, etc. Thes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/28B01J20/30G01N1/34
Inventor 廖且根罗林广
Owner INST OF AGRI PROD QUALITY SAFETY & STANDARD JIANGXI ACAD OF AGRI SCI
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