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Fast detecting method for aftatoxin B*

A technology for rapid detection of aflatoxins, applied in measuring devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of low detection accuracy, expensive instruments, and long time consumption, and achieve less organic reagents and other toxic reagents. High efficiency and recovery, and improved specificity and sensitivity

Inactive Publication Date: 2007-06-20
BEIJING CHINAINVENT INSTR TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, thin-layer chromatography is the most commonly used detection method for detecting aflatoxins. It does not require special equipment and can be carried out in general laboratories, but the amount of reagents is large, the operation is cumbersome, the interference of other components is serious, and the accuracy is poor. Quantitative, and more harmful to experimenters and the environment, not suitable for on-site rapid detection
Precision instrument analysis methods include fluorescence spectrophotometry and high performance liquid chromatography, which have high sensitivity and good accuracy, but the instruments are expensive, requiring a high degree of purification of aflatoxin samples, and the sample pretreatment process is cumbersome and time-consuming. High, difficult to achieve rapid detection
Immunological analysis methods include enzyme immunoassay, radioimmunoassay and time-resolved fluorescence method. Enzyme immunoassay has the advantages of strong specificity, high sensitivity, low cost, and is suitable for batch detection. However, it is generally a qualitative or semi-quantitative method. The detection of aflatoxins in agricultural products with complex components is susceptible to interference, and there are disadvantages such as a high proportion of false positives and low detection accuracy
Radioimmunoassay is similar to enzyme immunoassay, but radioimmunoassay still has radioactive pollution, which is likely to bring additional adverse effects on operators and the environment, and is rarely used now
Time-resolved fluorescence method has been applied in recent years, and its detection sensitivity is higher than the former two, but the pre-processing is still complicated, and the required equipment is expensive, so it is limited to the use in the field of medical clinical diagnosis

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  • Fast detecting method for aftatoxin B*

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: take by weighing 10g ground rice sample and put into stopper test tube, add 20ml concentration and be 50~60% methyl alcohol sodium chloride aqueous solution, contain 1~2% sodium chloride by weight in the methyl alcohol sodium chloride aqueous solution, Ultrasonic extraction in a water bath at 50°C for 3 minutes, then suction filtration with medium-speed qualitative filter paper, and extract 10ml of filtrate; add 2 to 4ml of redistilled petroleum ether to the filtrate, shake and mix well, and take 4ml of the lower layer solution after standing for 2 minutes. Add 20ml of pure water to the removed lower layer solution, and then pass the aflatoxin B 1 Immunoaffinity microcolumns, the aflatoxin B 1 The specification of the glass microcolumn used in the immunoaffinity microcolumn is Φ2~4mm (inner diameter)×60mm (length), and each gram of amino-modified silica gel particles (microspheres) contains 1.0~3.0 mg of aflatoxin B 1 Polyclonal antibody; wash aflatoxin B...

Embodiment 2

[0017] Embodiment 2: take by weighing 8.0g ground Shandong big peanut sample and put into stopper test tube, add 25ml concentration and be 80% methanolic sodium chloride aqueous solution, contain 1.8% sodium chloride by weight in methanolic sodium chloride aqueous solution, in Ultrasonic extraction in a water bath at 60°C for 5 minutes, suction filtration with medium-speed qualitative filter paper, and 5ml of filtrate; add 3ml of redistilled petroleum ether to the filtrate, shake and extract, and after standing for 2 minutes, take 1ml of the lower layer solution, and add the lower layer solution taken out to 4ml of pure water, then aflatoxin B 1 Immunoaffinity microcolumn, wash aflatoxin B with 10ml concentration of 10% aqueous methanol 1 The immunoaffinity micro-column was eluted twice with 2.0ml of pure methanol, and the eluate was collected, and 1.0ml of test reagent A was added, and then measured by a fluorescence spectrophotometer. According to aflatoxin B 1 Fluorescenc...

Embodiment 3

[0018] Embodiment 3: take by weighing 2.0g vegetable oil sample and put into tool stopper test tube, add 10ml concentration and be 70% methanolic sodium chloride aqueous solution, contain 1.5% sodium chloride by weight in methanolic sodium chloride aqueous solution, in 40~60 ℃ water bath Medium-ultrasonic extraction for 3 minutes, suction filtration with medium-speed qualitative filter paper, and 8ml of filtrate; add 4ml of redistilled petroleum ether to the filtrate, shake and extract, take 2ml of the lower layer solution after standing for 2-3 minutes, and add 8ml of ultrapure water, then aflatoxin B 1 Immunoaffinity microcolumn, wash aflatoxin B with 5ml concentration of 10% methanol water 1 The immunoaffinity micro-column was then eluted with 1.0ml of pure methanol, the eluate was collected, 0.5ml of test reagent A was added, and measured by a fluorescence spectrophotometer. According to aflatoxin B 1 Fluorescence spectrophotometer was used to measure the standard soluti...

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Abstract

The present invention is fast detection method of aflatoxin B1 in food and feed. The fast detection method includes the following steps: adding ground sample to be tested in 2-10 g in test tube, adding methanol-sodium chloride aqua of 50-80 % concentration in 10-25 ml, ultrasonic extraction in water bath at 40-60 deg.c, and suction filtering to obtain filtrate in 4-10 ml; adding re-distilled petroleum ether in 2-4 ml into the filtrate, shaking, letting stand to laminate, taking the lower layer of solution in 1-4 ml and adding pure water in 4-20 ml; adding to micro immunoaffinity column of aflatoxin B1, washing with methanol aqua of 10-40 % concentration in 5-10 ml, eluting with methanol in 0.5-2.0 ml, collecting the eluted liquid, adding test agent A in 0.2-1.0 ml; and detecting in fluorescent spectrophotometry or similar instrument. The present invention has the features of simple process, high detection precision and stability, fast detection speed and high safety.

Description

technical field [0001] The present invention relates to a kind of aflatoxin B in food and feed 1 quick test method. Background technique [0002] Mycotoxins are secondary metabolites secreted by toxin-producing fungi, and are natural toxic compounds that can cause various damages to humans and animals. Among the mycotoxins that have been discovered, aflatoxin B 1 (aflatoxin B 1 , referred to as AFB 1 ) is the most toxic mycotoxin, and its toxicity, carcinogenicity and pollution frequency all rank first in biological toxins. After it pollutes food and feed, it directly or indirectly enters the human food chain, threatening human health and life safety, and the degree of harm is proportional to the intake of aflatoxin. Aflatoxin widely exists in agricultural products such as rice, corn, peanuts, sesame, soybeans, rapeseed, fish and other foods, and there are many links in which it contaminates food and feed. 1 The maximum allowable content in food and feed is used as a m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/569G01N33/531G01N21/64
Inventor 李培武张文丁小霞杨金娥谢立华杨湄李光明
Owner BEIJING CHINAINVENT INSTR TECH
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