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Method for detecting illegal cooking oil, test paper and application of test paper

A technology of gutter oil and test strips, applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of false positives, low specificity, and low accuracy

Inactive Publication Date: 2012-07-18
GUANGZHOU CENT FOR DISEASE CONTROL & PREVENTION (GUANGZHOU HYGIENE INSPECTION CENT GUANGZHOU CENT FOR FOOD SAFETY RISK SURVEILLANCE & ASSESSMENT INST OF PUBLIC HEALTH OF GUANGZHOU MEDICAL UNIV)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, thin layer chromatography is prone to false positives for some unpurified oils (such as sesame oil, pepper oil, etc.) and oils with other ingredients (such as chili oil)
[0008] In summary, the existing methods are not very accurate and specific
On October 12, 2011, the spokesperson of the Ministry of Health of my country, Deng Haihua, revealed that the 5 detection methods for gutter oil collected so far are not very specific, and the relevant departments will open the collection methods to the public again.

Method used

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  • Method for detecting illegal cooking oil, test paper and application of test paper
  • Method for detecting illegal cooking oil, test paper and application of test paper
  • Method for detecting illegal cooking oil, test paper and application of test paper

Examples

Experimental program
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Embodiment 1

[0076] The preparation of embodiment 1 antibody A and antibody B

[0077] Antibody A and Antibody B both belong to monoclonal antibodies against the same substance, i.e. bacteria (Klebsiella pneumoniae) lipopolysaccharide or fat-soluble protein monoclonal antibodies, and the specific preparation method is as follows:

[0078] (1) Synthesis of bacterial lipopolysaccharide and liposoluble protein artificial immune antigen

[0079] After mixing methylated bovine serum albumin (BSA), glutaraldehyde, bacterial (Klebsiella pneumoniae) lipopolysaccharide and bacterial liposoluble protein in a molar ratio of 1:25:80, add 0.01M sodium carbonate The solution adjusts the pH of the mixture solution to 9.0, and reacts at 40°C for 6 hours, so that the bacterial lipopolysaccharide and fat-soluble protein are connected to the carrier protein, and the artificial immune antigen of the bacterial lipopolysaccharide and fat-soluble protein is obtained.

[0080] (2) Using bacterial lipopolysacchar...

Embodiment 2

[0093] Example 2 Preparation of fluorescein mixed label binding pad

[0094] 1. Preparation of dextran-antibody B-fluorescein mixed label

[0095] (1) Preparation of fluorescein-bound dextran molecules: Weigh 10 mg of fluorescein isothiocyanate (FITC), dissolve it in 10 mL of absolute ethanol, and add it dropwise to 20 mL of dextran under magnetic stirring conditions. In 10 mg / ml acetic acid solution (0.2 mol / L), react in the dark for 4 hours to make the carbon atoms on FITC react with divinyl sulfone on dextran for label coupling. Adjust the pH value to 9 with 10mol / L NaOH, centrifuge at 10000rpm for 5min, and take the precipitate.

[0096] (2) Preparation of primary product of dextran-antibody B-fluorescein mixed label: wash the precipitate with distilled water until the filtrate is clear and colorless. Dissolve the precipitate with 2% acetic acid solution by mass percentage again, adjust the pH value to 5 with 10mol / L NaOH, and add 10mL antibody B protein solution (20mg / m...

Embodiment 3

[0100] Example 3 Preparation of high-sensitivity fluorescent detection test paper for waste oil

[0101] 1. Materials required for the production of a waste oil detection immunofluorescence test strip

[0102] ① Nitrocellulose membrane: imported from Millipore / NC of the United States, the specification of nitrocellulose membrane is 30cm×3m / HAHY00010:

[0103] The area of ​​nitrocellulose membrane required for one immunofluorescence test strip is: 2.5cm×0.9cm.

[0104] The total area of ​​nitrocellulose membrane required to produce 100,000 copies of high-sensitivity fluorescent quantitative test strips is:

[0105] 2.5cm×0.9cm×100,000=2.25×105cm 2 ;

[0106] Total rolls of nitrocellulose membrane required to produce 100,000 test strips:

[0107] 2.25×105cm 2 / 30×300cm 2 = 25 volumes.

[0108] ② Anti-bacterial lipopolysaccharide or fat-soluble protein monoclonal antibody required for detection of T-line:

[0109] The total amount of anti-bacterial lipopolysaccharide or f...

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Abstract

The invention discloses a method for detecting illegal cooking oil, test paper and application of the test paper. According to the method, a lipopolysaccharide antibody and / or a fat-soluble protein antibody are / is taken as an antibody for detection. The method comprises the following steps of: preparing an antibody A and an antibody B by using a hybridoma cell strain gzcdc-ETX001; reacting fluorescein molecules with glucan molecules to obtain glucan molecules combined with fluorescein, reacting the glucan molecules combined with the fluorescein with the antibody B, and purifying to obtain a glucan-antibody B-fluorescein mixed marker; and reacting a sample to be tested with the antibody A, adding the glucan-antibody B-fluorescein mixed marker, continuing to react, and detecting by using a device for detecting fluorescence so as to judge whether the illegal cooking oil is contained in the sample to be tested. The glucan-antibody B-fluorescein mixed marker is enveloped by a bonding pad in the test paper for implementing the method; and a detection band on a chromatographic membrane is fixed with the antibody A, and a quality control band is fixed with an antiantibody which can be specifically combined with the glucan-fluorescein marked antibody B. The test paper is sensitive and easy to use.

Description

technical field [0001] The invention relates to the field of food safety detection and the technical field of immunofluorescence detection, in particular to a method for detecting waste oil, a test paper and its application. Background technique [0002] Waste oil is a non-edible oil with extremely poor quality and hygiene, and indicators such as peroxide value, acid value, moisture, carbonyl value, malondialdehyde, and AFBI (Aflatoxin Bl (AFBI)) seriously exceed the standard. Waste oil from the catering industry contains a large amount of toxic and harmful substances. After human consumption, it will cause poisoning symptoms such as dizziness, nausea, vomiting, diarrhea, insomnia, fatigue, indigestion, liver discomfort and severe abdominal cramps. Long-term consumption will cause anemia, Malnutrition and damage to internal organs can lead to weight loss and developmental disorders in children. In severe cases, toxic liver disease may occur, which can induce gastric adenocar...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 陈守义杨智聪李军涛侯水平姬泽薇
Owner GUANGZHOU CENT FOR DISEASE CONTROL & PREVENTION (GUANGZHOU HYGIENE INSPECTION CENT GUANGZHOU CENT FOR FOOD SAFETY RISK SURVEILLANCE & ASSESSMENT INST OF PUBLIC HEALTH OF GUANGZHOU MEDICAL UNIV)
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