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Anti-capsaicin substance general specific antibody, test paper strip and kitchen waste oil immunochromatographic rapid discriminating method

A technology for waste oil and capsaicin, applied in the field of immunochemistry, can solve the problems of difficulty in determining the pH of the sample extract, reduced specific antibody activity, and reduced detection sensitivity.

Active Publication Date: 2016-11-09
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 in kitchen waste oils and fats at home and abroad.
The core component of colloidal gold immunochromatography technology is specific antibody, but the matrix composition of kitchen waste oil is complex, and the pH of the sample extract is difficult to determine, which usually reduces the activity of specific antibody and the detection sensitivity

Method used

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  • Anti-capsaicin substance general specific antibody, test paper strip and kitchen waste oil immunochromatographic rapid discriminating method
  • Anti-capsaicin substance general specific antibody, test paper strip and kitchen waste oil immunochromatographic rapid discriminating method
  • Anti-capsaicin substance general specific antibody, test paper strip and kitchen waste oil immunochromatographic rapid discriminating method

Examples

Experimental program
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Effect test

Embodiment 1

[0064] The preparation of embodiment 1 capsaicinoid artificial hapten

[0065] (1) Weigh 1 g of vanillin amine hydrochloride and dissolve it in 11.77 ml of ultrapure water, add 2.12 ml of 2M NaOH drop by drop under vigorous stirring, and react at room temperature for 10 min. The white precipitate was obtained by filtration, and the next reaction was carried out after the white precipitate was thoroughly dried.

[0066] (2) Weigh and weigh 60 mg of the product from the previous step, dissolve it in 30 ml of chloroform (after dehydration with anhydrous sodium sulfate), stir at room temperature for 40 min, add 40 mg of maleic anhydride, and react at room temperature for 1 h. The resulting precipitate after filtration is capsaicinoid artificial hapten (2Z)-4-[(4-hydroxy-3-methoxy)benzylamino]-4-carbonyl-2-enoic acid ((2Z)-4- [(4-hydroxy-3-methoxybenzyl)amino]-4-oxobut-2-enoic acid). Molecular formula is C 12 h13 NO 5 .

[0067]

Embodiment 2

[0068] Example 2 Preparation of capsaicinoids artificial complete antigen-immune antigen

[0069] Weigh 20.8mg (about 0.08mmol) of the artificial hapten of the above-mentioned capsaicinoids and 11.6mg (about 0.1mmol) of NHS in the reaction flask, add 400ulDMF to dissolve in the reaction flask, stir at room temperature for 30min, weigh 20.6mg (about 0.1mmol) mmol) DCC was dissolved in 50 ul of DMF, and the DCC / DMF solution was added dropwise to the above-mentioned reaction flask, stirred at room temperature for 4 h, and left standing at 4° C. overnight. 8000rpm / 5min, take the supernatant active ester solution.

[0070] The supernatant active ester solution was added dropwise to 6ml of 7mg / ml BSA solution for reaction, and the reaction buffer was 0.2M phosphate buffer at pH 8.0. The reaction was carried out at room temperature for 4 h under magnetic stirring. The reaction solution was placed in a dialysis bag, and dialyzed with 0.01M PBS, pH 7.4 at 4°C with stirring, and the d...

Embodiment 3

[0071] Example 3 Preparation of capsaicinoids artificial complete antigen-coated antigen

[0072] Dissolve 4.52 mg of the above artificial hapten of capsaicinoids in 200 μL of anhydrous DMF, then add 4.27 μL of tri-n-butylamine and 2.34 μL of isobutyl chloroformate in sequence, and stir at room temperature for 1 hour in the dark to obtain activated capsicum. Capsaicin, dihydrocapsaicin and synthetic capsaicin hapten solutions.

[0073] The activated capsaicin, dihydrocapsaicin and synthetic capsaicin hapten solutions were added dropwise to 10ml of 4.5mg / ml OVA solution for reaction, and the reaction buffer was 0.2M phosphate buffer at pH 8.0. The reaction was carried out at room temperature for 4 h under magnetic stirring. The reaction solution was placed in a dialysis bag and dialyzed with 0.01M PBS with pH 7.4 at 4°C with stirring, and the dialysate was changed every 4 hours for a total of 72 hours. That is, capsaicin artificial antigen-coated antigen is obtained. For the...

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Abstract

The invention relates to an anti-capsaicin substance general specific antibody, a test paper strip and a kitchen waste oil immunochromatographic rapid discriminating method. The anti-capsaicin substance general specific antibody is obtained through immunizing animals with a capsaicin substance general artificial immunization antigen compound represented by formula I, separating and purifying, and freeze-drying. The kitchen waste oil immunochromatographic immune test paper strip made from the anti-capsaicin substance general specific antibody can be used for rapidly detecting the total amount of capsaicin substances in kitchen waste oil, and has the advantages of high detection sensitivity, wide pH adaptation range, one-step operation, and high simplicity and convenience. No capsaicin substance standard solutions are used as positive control substances.

Description

technical field [0001] The invention relates to a general-specific specific antibody against capsaicinoids, a test strip and a rapid immunochromatographic identification method for kitchen waste oil, belonging to the technical field of immunochemistry. Background technique [0002] Waste oil is a general concept, which is a general term for all kinds of low-quality oil, generally including swill oil, frying waste oil, food and waste oil produced by related enterprises, and swill oil from the catering industry is the main raw material source of waste oil one. In Chinese food culture, pepper containing capsaicin is one of the essential condiments. Capsaicinoids are the main chemical substances present in capsicum fruits that make them pungent and stimulating. There are more than 20 kinds of compounds whose structures have been proven so far. Capsaicinoids have high nutritional value and are widely used in the field of food processing. Synthetic capsaicin also has a structur...

Claims

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Application Information

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IPC IPC(8): C07K16/44C07K16/06G01N33/92G01N33/558
CPCG01N33/558G01N33/92C07K16/06C07K16/44G01N33/03G01N33/5308C07K14/765C07K19/00C07K2317/30
Inventor 李培武杨青青张奇马飞张良晓张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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