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A test paper card for detecting aflatoxin b1 and its application

A technology of aflatoxin and colloidal gold test paper, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that are not suitable for large-scale sample detection, sample purity requirements are not high, and are not suitable for rapid detection, etc., and the detection time is short , low cost, easy to promote and use

Active Publication Date: 2016-03-30
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography has high sensitivity, strong separation ability, and good specificity, etc., but some samples need to be thoroughly and effectively pre-treated, which is not suitable for the detection of large-scale samples, and the equipment is expensive and difficult to popularize
Enzyme-linked immunosorbent assay is a commonly used detection method at present. It has the advantages of rapidity, sensitivity, quantification, and low requirements on sample purity. It is especially suitable for the detection of large batches of samples. The personnel and the detection time are relatively long, so it is not suitable for on-site rapid detection

Method used

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  • A test paper card for detecting aflatoxin b1 and its application
  • A test paper card for detecting aflatoxin b1 and its application
  • A test paper card for detecting aflatoxin b1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation of embodiment 1 aflatoxin B1 detection test paper card

[0041] The preparation method of the test paper card mainly comprises the following steps:

[0042] 1) Prepare a conjugate release pad sprayed with aflatoxin B1 monoclonal antibody-colloidal gold marker;

[0043] 2) Prepare a reaction membrane with a detection line coated with aflatoxin B1 hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;

[0044] 3) Assemble the conjugate release pad, reaction membrane, sample absorbent pad, water absorbent pad and bottom plate prepared in 1) and 2) into a test strip card.

[0045] The following step-by-step detailed description:

[0046]1. Preparation of aflatoxin B1 hapten

[0047] Add the mixture of 0.10g aflatoxin B1 in 2ml dimethylsulfoxide (DMSO) slowly at 60°C to the mixture of 0.1ml 1,3-propanediamine and 0.1ml pyridine in 2ml DMSO, after the addition is complete , continue the reaction for 12h, remo...

Embodiment 2

[0079] The detection of aflatoxin B1 residue in the sample of embodiment 2

[0080] 1. Sample pretreatment

[0081] Weigh 3.0g±0.05g crushed sample into a 15ml or 50ml polystyrene centrifuge tube, add 6ml of acetonitrile, cover the bottle tightly, shake for 5min; centrifuge at room temperature (20-25℃) over 3000g for 5min; Put the supernatant into a glass centrifuge tube, dry it in a water bath at 50-60°C under nitrogen flow, add 0.4ml of sample complex solution (0.02mol / L phosphate buffer), vortex for 30s, and wait for detection.

[0082] 2. Test with test paper card

[0083] Use a straw to absorb the sample solution to be tested and add 3 drops vertically to the sample hole, start timing when the liquid flows, react for 5-10 minutes, and judge the result. After 15 minutes, it is judged invalid.

[0084] 3. Analyze test results

[0085] Negative (-): Both the T line and the C line are colored, indicating that the concentration of aflatoxin B1 in the sample is lower than t...

Embodiment 3

[0088] Example 3 sample detection example

[0089] 1. Detection limit test

[0090] Take blank grains (soybeans, corn) and feed (raw materials, compound materials, concentrates), add aflatoxin B1 to them to a final concentration of 2.5, 5, and 10 μg / kg, take a test paper card for detection, and repeat for each sample Measured three times.

[0091] When testing samples of soybeans, corn, raw materials, batch materials, and concentrated materials with a test paper card, when the concentration of aflatoxin B1 is 2.5 μg / kg, two red lines visible to the naked eye are displayed on the test paper card, which is negative; Among them, when the added concentration of aflatoxin B1 is 5, 10 μg / kg, the quality control line of the test paper card develops color, but the detection line does not develop color, which is positive, indicating that the detection limit of this test paper card for aflatoxin B1 in grain and feed samples 5 μg / kg.

[0092] 2. False positive rate and false negative ...

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Abstract

The invention discloses a test paper card for detecting aflatoxin B1 and application of the test paper card. The test paper card comprises a sample absorption pad, a conjugate release pad, a reactive film, a water absorption pad and a bottom plate, wherein a detection line coated with an aflatoxin B1 hapten-carrier protein conjugate and a quality control line coated with a goat anti-rat antibody are arranged on the reactive film; an aflatoxin B1 monoclonal antibody-colloid gold marker is sprayed on the conjugate release pad. The invention also provides a method for detecting aflatoxin B1 residues in grains and feed by using the aflatoxin B1 test paper card. The test paper card provided according to the invention has the characteristics of simplicity in operation, high sensitivity, high detection speed, low cost and the like and is suitable for screening and field monitoring of lots of samples.

Description

technical field [0001] The invention relates to a test paper card for detecting aflatoxin B1 and its application, in particular to a colloidal gold test paper card for detecting aflatoxin B1, which is especially suitable for grains such as soybeans and corn and feed (raw materials, batch materials, etc.) , concentrate) detection of aflatoxin B1 residues in samples. Background technique [0002] Aflatoxin (AF) is the strongest type of biological toxin found to pollute agricultural products so far, and it is also a strong carcinogen. It is a class of metabolites produced by various fungi such as Aspergillus flavus and A. parasiticus, a group of compounds with similar structures. It is easy to naturally exist in crops such as peanuts, cottonseed, corn, wheat and rice, and has strong toxicity and carcinogenicity. It is the strongest carcinogenic mutagen among chemical carcinogens, and its toxicity is 10 times stronger than potassium cyanide. times, its carcinogenicity is 75 ti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/531
CPCG01N33/531G01N33/558G01N33/577
Inventor 何方洋冯才伟崔海峰冯才茂陶光灿彭鸽马腊腊王建霞
Owner BEIJING KWINBON BIOTECH
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