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Molecularly imprinted polymer test strip for detecting 17β-estradiol and preparation method thereof

A technology of molecular imprinting and estradiol, which is applied in biological testing, chemical instruments and methods, analytical materials, etc., can solve the problems of easy failure, complicated process and high cost

Active Publication Date: 2019-03-19
山东师大瑞柏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods require complex pretreatment of samples during actual operation, which is expensive and complicated, and is not suitable for real-time on-site determination.
Rapid detection methods are mainly immunological methods, including enzyme-linked immunosorbent assays and test strip methods. This method has high selectivity due to the specific combination of antigens and antibodies, but due to the disadvantages of difficult screening, storage, and failure of antibodies, such as Chinese patent "a kit for detecting estradiol using magnetic particle chemiluminescence immunotechnology", application number: 200910091848.9, which discloses a kit for detecting estradiol using magnetic particle chemiluminescence immunotechnology

Method used

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  • Molecularly imprinted polymer test strip for detecting 17β-estradiol and preparation method thereof
  • Molecularly imprinted polymer test strip for detecting 17β-estradiol and preparation method thereof
  • Molecularly imprinted polymer test strip for detecting 17β-estradiol and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Synthesis of 17β-estradiol-HRP

[0062] The 17β-estradiol used in this example was purchased from Shanghai Ruiyong Biotechnology Co., Ltd., and HRP was purchased from sigma-aldrich (Shanghai) Trading Co., Ltd.

[0063] Proceed as follows:

[0064] (1) Synthesis of 17β-estradiol-3-carboxymethyl ether:

[0065] Dissolve 300mg 17β-estradiol in 6mL dry dimethyl sulfoxide, add 1g potassium hydroxide powder, stir for 5 minutes, add 300mg bromoacetic acid, continue stirring, react for 2 hours, add ice water; ethyl acetate extraction and recovery For the unreacted 17β-estradiol, the aqueous phase was acidified with 2mol / L hydrochloric acid, and a white precipitate appeared, which was filtered through a funnel. The precipitate was washed with deionized water to neutrality, and dried in a vacuum of 0.08MPa.

[0066] (2) Coupling of 17β-estradiol-3-carboxymethyl ether with HRP:

[0067] ① Weigh 330 mg of 17β-estradiol-CME, 145 mg of N-hydroxysuccinamide, and 260 mg of dicycloethylcarbodiimi...

Embodiment 2

[0070] Synthesis of 17β-estradiol-HRP

[0071] Proceed as follows:

[0072] (1) Synthesis of 17β-estradiol-3-carboxymethyl ether:

[0073] Dissolve 1g 17β-estradiol in 10mL dry dimethyl sulfoxide, add 5g potassium hydroxide powder, stir for 5 minutes, add 1g bromoacetic acid, continue stirring, react for 2 hours, add ice water; ethyl acetate extraction and recovery For unreacted 17β-estradiol, the aqueous phase was acidified with 2mol / L hydrochloric acid, and a white precipitate appeared, which was filtered through a funnel. The precipitate was washed with deionized water to neutrality, and dried in a vacuum of 0.08MPa.

[0074] (2) Coupling of 17β-estradiol-3-carboxymethyl ether with HRP:

[0075] ① Weigh 1g 17β-estradiol-CME, 0.8g N-hydroxysuccinamide, 0.5g dicycloethylcarbodiimide, add 80mL dimethyl sulfoxide, and stir for 2 hours.

[0076] ②8mg HRP is dissolved in 5mL sodium bicarbonate solution, and the solution ① is slowly dropped into the HRP solution and stirred gently for 2 hou...

Embodiment 3

[0078] Synthesis of molecularly imprinted polymer test strips:

[0079] The paper used in this embodiment is INYC00010 Charged nylon positively charged nylon membrane NY+ purchased from Millipore Corporation (Billerica, USA).

[0080] Proceed as follows:

[0081] (1) Activation test strip

[0082] 100mg 3-(methacryloyloxy)propyltrimethoxysilane is dissolved in 10mL ethanol solution, and then the blank test strip is immersed in the prepared solution and incubated at room temperature for 1 hour. The excess silane can be passed with ethanol Rinse the solution to remove it, and then place the treated test strip in an air stream to dry it.

[0083] (2) Synthesis of molecularly imprinted polymers on test strips:

[0084] (1) The polymerization solution contains 0.1g 17β-estradiol (template molecule), 1014μL 3-aminopropyltriethoxysilane (APTES, functional monomer), 963μL tetraethoxysilane (TEOS, crosslinking agent) , 0.1 mol / L acetic acid (catalyst) was dissolved in 12 mL of acetonitrile solu...

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Abstract

The invention discloses a molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum and a preparing method thereof. The test strip is modified with the molecularly imprinted polymer, and the molecularly imprinted polymer bears specific binding sites of 17 beta-Estradiolum. The specific binding sites can not only carry out specific binding with enzyme marked 17 beta-Estradiolum, but also can carry out specific binding with 17 beta-Estradiolum. The detection is realized through the competitive binding between the 17 beta-Estradiolum to be detected, the enzyme marked 17 beta-Estradiolum and the biding sites. The molecularly imprinted polymer test strip used for detecting 17 beta-Estradiolum can realize the specific binding of the target template module and can magnify signal through the color reaction between enzyme and substrate. The method for detecting 17 beta-Estradiolum does not need complicated sample pretreatment, has simple operation, low cost, and good reliability, and can be used for the detection of 17 beta-Estradiolum in food or clinic.

Description

Technical field [0001] The invention belongs to the technical field of molecular imprinting, and specifically relates to a molecularly imprinted polymer test strip for detecting 17β-estradiol, and a preparation method and detection method thereof. Background technique [0002] Estradiol (estradiol, C 18 H 24 O 2 ) Is a steroidal estrogen, there are two types of α, β, 17β-estradiol (17β-estradiol, 17β-E 2 ) Is one of the most active substances in environmental estrogen. Due to the obvious carcinogenicity of 17β-estradiol, countries such as Europe and the United States have successively banned or strictly banned its use, but 17β-estradiol has a growth-promoting effect. Abuse is widespread in developing countries, and there is a high probability of residues in animal food. In addition, 17β-estradiol can be enriched in organisms through the food chain in the ecosystem, causing a long-term adverse effect and posing a serious threat to the health of humans and natural organisms. The c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G77/26C08G77/08G01N33/74G01N33/531B01J20/26B01J20/30
CPCB01J20/268C08G77/08C08G77/26G01N33/531G01N33/74G01N2600/00
Inventor 张鸿雁肖蕾刘兆臣张震
Owner 山东师大瑞柏生物科技有限公司
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