A time-resolved fluorescent immunochromatographic test strip for detecting coumarin and its preparation method and application

An immunochromatography test paper and time-resolved fluorescence technology, which is applied to the analysis of materials, measuring devices, instruments, etc., can solve the problems of high professional requirements for operators, expensive detection costs, cumbersome steps, etc., and achieve a wide range of applications and detection The effect of short time and simple operation

Active Publication Date: 2020-09-22
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the above methods require advanced detection instruments, the detection costs are expensive, the steps are cumbersome, time-consuming, and require high professionalism of operators, they are not suitable for high-throughput rapid screening detection in grassroots enterprises and institutions.

Method used

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  • A time-resolved fluorescent immunochromatographic test strip for detecting coumarin and its preparation method and application
  • A time-resolved fluorescent immunochromatographic test strip for detecting coumarin and its preparation method and application
  • A time-resolved fluorescent immunochromatographic test strip for detecting coumarin and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 The composition of the time-resolved fluorescent immunochromatographic test strip for detecting coumarin

[0041] 1. Test strips

[0042] The test strip is composed of a bottom plate, a sample absorbent pad, a conjugate release pad, a nitrocellulose membrane and an absorbent pad;

[0043] The sample absorbent pad 1, the conjugate release pad 2, the nitrocellulose membrane 3 and the water absorbent pad 4 are overlapped and pasted on the bottom plate 7 in sequence, and 1 / 3 of the conjugate release pad is covered by the sample absorbent pad from the starting end , the end of the conjugate release pad is connected to the beginning of the nitrocellulose membrane, the end of the nitrocellulose membrane is connected to the beginning of the absorbent pad, the beginning of the sample absorbent pad is aligned with the beginning of the PVC bottom plate, and the end of the absorbent pad is connected to the end of the PVC bottom plate alignment;

[0044] The nitrocellulo...

Embodiment 2

[0046] Example 2 Preparation of time-resolved fluorescent immunochromatographic test strips for detection of coumarin

[0047] The preparation method of the time-resolved fluorescent immunochromatographic test strip for detecting coumarin mainly comprises the following steps:

[0048] 1) Preparation of the conjugate release pad: label the coumarin monoclonal antibody with commercially available fluorescent microspheres and dilute it with a specific buffer system, soak the conjugate release pad in the dilution buffer, and freeze-dry in vacuum Post-preparation;

[0049] 2) Preparation of nitrocellulose membrane: Spray the coumarin hapten-carrier protein conjugate on the detection area of ​​the nitrocellulose membrane to make the detection area; spray goat anti-mouse anti-antibody onto the nitrocellulose membrane The scope of the quality control area above is made into the quality control area;

[0050] 3) Assembly and cutting: Paste the sample absorption pad, the conjugate rel...

Embodiment 3

[0103] Example 3 Application of time-resolved fluorescent immunochromatographic test strips for detection of coumarin

[0104] 1. Tobacco sample pretreatment

[0105] Weigh 1.0±0.05 g of crushed tobacco sample into a polystyrene centrifuge tube; add 10 mL of 50% methanol aqueous solution, and use a homogenizer to fully break it up to obtain a sample liquid; pipette 75 μL of sample liquid and 425 μL of The ionized water was mixed and waited for inspection.

[0106] 2. Test with test strips

[0107] Draw 100 μL of the sample solution to be tested and drop it vertically into the sample hole of the test paper card. When the liquid flows, start timing and react for 10 minutes; insert the test paper card into the carrier of the KFT-100A fluorescence detector, and select the sample to be Check the items, press the "Start Test" button, the fluorescence detector will automatically scan the test paper card, and read or print the test results on the display screen of the instrument.

...

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Abstract

The invention discloses a time resolved fluorescence immunochromatography test strip for detecting coumarin as well as a preparation method and application thereof. The test strip comprises a bottom plate, a sample absorption pad, a compound release pad, a nitrocellulose membrane and a water absorption pad, wherein the sample absorption pad, the combined object release pad, the nitrocellulose membrane and the water absorption pad are sequentially lapped and stuck; a fluorescent microsphere marked coumarin monoclonal antibody is embedded on the compound release pad; a detecting region and a quality control region are fixed on the nitrocellulose membrane; a coumarin hapten-carrier protein conjugate is sprayed and coated on the detecting region; an anti-goat anti-mouse antibody is sprayed andcoated on the quality control region; the coumarin hapten is obtained from coumarin through aldolization. The invention also provides the preparation method of the test strip and a method for using the coumarin in a test strip detecting sample. The test strip and the detecting method provided by the invention have the advantages of operation simplicity, high sensitivity, high detecting speed andlow cost; the fast detection and field monitoring on coumarin in a great batch of samples can be realized.

Description

technical field [0001] The invention belongs to the field of detection of illegal additives in food safety, and in particular relates to a time-resolved fluorescent immunochromatographic test strip for detecting coumarin in tobacco flavors and fragrances, tobacco and tobacco products, and a preparation method and application thereof. Background technique [0002] Coumarin-based compounds are a class of natural products with an aromatic odor and a benzoα-pyrone structure as the core. They are widely distributed in the plant kingdom and can be divided into simple coumarin-based compounds and furanocoumarin-based compounds according to their structures. Compounds, pyranocoumarin-based compounds and other coumarin-based compounds, etc. Coumarin, also known as o-hydroxycinnamic acid lactone and 1,2-benzopyrone, is a natural plant secondary product and a lactone compound widely present in nature. It was discovered in 1820 Found in tonka bean, mainly exists in plants in the form o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/558G01N33/577
CPCG01N33/533G01N33/558G01N33/577
Inventor 陈黎刘惠民崔华鹏赵乐樊美娟谢复炜张晓兵聂聪秦亚琼尚平平
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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