Colloidal gold immunochromatography test strip for rapidly detecting capsaicinoids as well as preparation method and application thereof

An immunochromatographic test strip and capsaicin technology, which is applied in biological tests, measuring devices, analytical materials, etc., can solve the problems of blurred spiciness level labels, and achieve the effects of simple operation and high detection sensitivity.

Active Publication Date: 2016-05-04
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, this method has not been applied to the detection of capsaicinoids at home and abroad.
Due to the eating habits of our country, chili food is abundant, but the hotness level in the product description...

Method used

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  • Colloidal gold immunochromatography test strip for rapidly detecting capsaicinoids as well as preparation method and application thereof
  • Colloidal gold immunochromatography test strip for rapidly detecting capsaicinoids as well as preparation method and application thereof
  • Colloidal gold immunochromatography test strip for rapidly detecting capsaicinoids as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1 Obtaining of anti-capsaicinoid monoclonal antibody

[0047] The anti-capsaicinoid monoclonal antibody is a monoclonal antibody secreted by the hybridoma cell line YQQD8 with the preservation number CCTCCNO.C201534. The specific preparation method:

[0048] The hybridoma cell line YQQD8 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. Rat ascites, 4°C, centrifuged at 12000r / min for 30min, sucked the supernatant, mixed the obtained ascites supernatant with 3 times the volume of acetate buffer, slowly added n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites was 33μL, mix at room temperature for 30min, stand at 4°C for more than 2h; centrifuge at 12000r / min for 30min at 4°C, discard the precipitate, filter the obtained supernatant with double-layer filter paper, add 1 / 10 o...

Embodiment 2

[0075] Embodiment 2: the preparation method of the colloidal gold immunochromatography test paper strip of rapid detection capsaicinoid, the steps are as follows:

[0076] (1) Preparation of absorbent pad

[0077] Cut the absorbent paper to a length of 16mm and a width of 4mm to obtain an absorbent pad;

[0078] (2) Preparation of detection pad

[0079] Coating of the detection line: the capsaicin-like substance-coated antigen, that is, the above-mentioned capsaicin-like substance general-purpose artificial complete antigen-coated antigen, is prepared into a coating solution with a concentration of 0.5 mg / mL, and placed on the upper edge of the nitrocellulose membrane. At the position of 15mm, it was coated horizontally on the nitrocellulose membrane by line spraying method to obtain the detection line. The coating amount of capsaicinoid-coated antigen required for each centimeter of detection line was 0.3 μg, and then at 37°C Dry for 30 minutes;

[0080] The coating buffer...

Embodiment 3

[0100] Embodiment 3: the preparation method of the colloidal gold immunochromatography test strip of rapid detection capsaicinoid substance, the steps are as follows:

[0101] (1) Preparation of absorbent pad

[0102] Cut the absorbent paper to a length of 18mm and a width of 4mm to obtain an absorbent pad;

[0103] (2) Preparation of detection pad

[0104] Coating of the detection line: the capsaicin-like substance-coated antigen, that is, the above-mentioned capsaicin-like substance universal artificial complete antigen-coated antigen, is prepared into a coating solution with a concentration of 0.25 mg / mL, and placed on the edge of the nitrocellulose membrane. At the position of 9mm, it was coated horizontally on the nitrocellulose membrane by line spraying method to obtain the detection line. The coating amount of capsaicinoid-coated antigen required for each centimeter of detection line was 0.15 μg, and then at 37°C Dry for 20 minutes;

[0105] The coating buffer used i...

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Abstract

The invention relates to a colloidal gold immunochromatography test strip for rapidly detecting capsaicinoids as well as a preparation method and application thereof. The colloidal gold immunochromatography test strip is characterized by comprising a paperboard, wherein a water absorption pad, a detection pad, a gold mark pad and a sample pad are stuck on one face of the paperboard in sequence from top to bottom; the adjacent pads are overlapped and connected at a connection part; the detection pad is used as a base pad of a nitrocellulose membrane; the nitrocellulose membrane is provided with a transverse quality control line and a detection line from top to bottom; a rabbit anti-mouse polyclonal antibody covers the quality control line; a capsaicinoids coating antigen coats the detection line; the gold mark pad is transversely sprayed with a nano gold marked common monoclonal antibody resisting capsaicin, dihydrocapsaicin and synthesized capsaicin; and the nano gold marked common monoclonal antibody resisting capsaicin, dihydrocapsaicin and synthesized capsaicin are produced by secretion of hybridoma cell strains YQQD8 with the preservation number of CCTCC (China Center For Type Culture Collection) NO.C201534. The colloidal gold immunochromatography test strip for rapidly detecting the capsaicinoids, provided by the invention, can be used for simultaneously detecting the total content of the capsaicin, the dihydrocapsaicin and the synthesized capsaicin; and the detection sensitivity is high and the minimum detection limit is 10ng/mL.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a colloidal gold immunochromatographic test strip for rapidly detecting capsaicin substances, a preparation method and an application thereof. Background technique [0002] Capsaicin compounds are the main chemical substances that make peppers pungent, and their components mainly include capsaicin, dihydrocapsaicin, homodihydrocapsaicin, nordihydrocapsaicin, homocapsaicin and other compounds. Wherein capsaicin and dihydrocapsaicin account for about 90% of the total amount of capsaicinoids, and provide about 90% of the hot stimulation. In addition to giving food a spicy taste and being used as a condiment, capsaicinoids also have anti-inflammatory and analgesic, anti-bacterial, anti-tumor effects, can promote fat burning, lower blood lipids, inhibit gastric acid secretion, and have protective effects on gastric mucosal damage. Use value. In the field of food proce...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/52
CPCG01N33/52G01N33/558G01N33/577
Inventor 李培武杨青青马飞张奇张良晓张文丁晓霞张兆威
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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