Detection method of aflatoxin b1

A technology of aflatoxin and detection method, applied in the field of detection of aflatoxin B1, can solve the problems of long operation time, fluctuation of recovery rate, interference, etc., and achieve the effect of good stability, simple steps and rapid detection

Active Publication Date: 2018-03-02
GUANGDONG TESTING INST OF PROD QUALITY SUPERVISION
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, the enzyme-linked immunosorbent assay for the determination of aflatoxin in soy sauce and vinegar has been reported, and some related products are also available, but the pretreatment process of the samples is relatively complicated, and highly toxic organic solvents such as chloroform are used, and nitrogen is required at the same time. Blowing treatment, the reason is that the pigment and high salt content in soy sauce will interfere with the detection system and cause inaccurate detection results. Therefore, it is necessary to use organic solvents such as chloroform for extraction and nitrogen blowing reconstitution for measurement
However, such a pretreatment method takes a long time to operate, and the nitrogen blowing process needs to be strictly monitored, which can easily lead to fluctuations in the recovery rate.

Method used

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  • Detection method of aflatoxin b1
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  • Detection method of aflatoxin b1

Examples

Experimental program
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Effect test

Embodiment 1

[0048] A method for detecting aflatoxin B1 includes the following steps:

[0049] 1. Pre-treatment.

[0050] The sample to be tested is extracted with the extraction solution. In this embodiment, the sample to be tested is soy sauce.

[0051] The specific pre-treatment method is as follows: Weigh 5.0g representative sample into a 50mL polystyrene centrifuge tube, add different ratios of different extraction solutions and mix with them; vigorously shake on a shaker for 10 minutes at a speed of 150r / min (Or vortex for more than 5min); centrifuge the liquid at 4000r / min for 5min (or stand still for 3min, and then filter with quantitative filter paper); take 0.5mL of supernatant or filtrate, and then add diluent in different proportions to obtain the solution to be tested. Shake the solution to be tested for 5s or shake it by hand, and take 50μL for analysis.

[0052] 2. Testing.

[0053] Take the test solution, add it to the enzyme-labeled plate coated with aflatoxin B1 antigen, then add...

Embodiment 2

[0120] A method for detecting aflatoxin B1 includes the following steps:

[0121] 1. Pre-treatment.

[0122] The sample to be tested is extracted with the extraction solution. In this embodiment, the sample to be tested is bean paste.

[0123] The specific pretreatment method is as follows: Weigh 4.0g representative sample into a 50mL polystyrene centrifuge tube, add 10mL methanol as the extraction solution and mix with it; vigorously shake on a shaker for 10 minutes at a speed of 150r / min ( Or vortex for more than 5min); centrifuge the liquid at 4000r / min for 5min (or stand still for 3min, and then filter with quantitative filter paper); take 0.1mL of supernatant or filtrate, and add 1.9mL of deionized water as the diluent to obtain the test Solution, the dilution factor is 50. The solution to be tested is shaken for 5 seconds or shaken by hand, and 50 μL is taken for analysis.

[0124] 2. Testing.

[0125] The detection is carried out according to the method of claim 1.

[0126] 3. ...

Embodiment 3

[0134] A method for detecting aflatoxin B1 includes the following steps:

[0135] 1. Pre-treatment.

[0136] The sample to be tested is extracted with the extraction solution. In this embodiment, the sample to be tested is cooking wine, rice wine, pipa wine, and white vinegar.

[0137] The specific pre-treatment method is as follows: Weigh 5.0g representative sample into a 100mL Erlenmeyer flask with stopper, add 25mL of 60% methanol as the extraction solution and mix with it; vigorously shake on a shaker for 10 minutes at a speed of 150r / min ( Or vortex for more than 5min); centrifuge the liquid at 4000r / min for 5min (or stand still for 3min, and then filter with quantitative filter paper); take 1mL of supernatant or filtrate, and add 5mL of deionized water as the dilution to obtain the solution to be tested. The pH value of the solution to be tested should be between 6 and 8, and it can be adjusted with "NaOH" and "HCL". The solution to be tested is shaken for 5 seconds or shaken...

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Abstract

The invention relates to a detection method of aflatoxin B1 and belongs to the technical field of ELISA (enzyme-linked immunosorbent assay). The detection method comprises steps as follows: pretreatment: a to-be-detected sample is extracted with an extracting solution, then a diluent is added, and a to-be-detected solution is obtained, wherein the extracting solution is a methanol aqueous solution with the concentration of 40%-100% in percentage by volume and / or acetonitrile, and the diluent is water or a buffered saline solution; detection: the to-be-detected solution is taken and added to an ELISA plate coated with an aflatoxin B1 antigen, then an aflatoxin B1 antibody solution is added, incubation is performed, a horseradish peroxidase marked goat-anti-mouse second antibody is added for amplification of enzymatic activity, the plate is washed, a substrate color developing agent and a stop solution are added, an absorbance value is determined by a microplate reader, and the content of aflatoxin B1 in the to-be-detected sample is obtained through calculation. The detection method omits the pretreatment steps such as nitrogen blowing and the like and has the advantages of simple steps and good stability, and the residual amount of aflatoxin B1 can be rapidly detected.

Description

Technical field [0001] The invention relates to the technical field of enzyme-linked immunosorbent assay, in particular to a detection method of aflatoxin B1. Background technique [0002] At present, the detection methods of aflatoxin B1 in foods such as soy sauce and vinegar mainly include chemical analysis, instrumental analysis, immunoassay, and biosensor. [0003] Among them, the most commonly used chemical analysis method is thin layer chromatography, and thin layer chromatography (TCL) uses the characteristics of certain mycotoxins (such as aflatoxins) that can display fluorescence at a specific wavelength. This method is more economical and does not require high equipment and inspection personnel, but it cannot be accurately quantified. [0004] Instrumental analysis methods mainly include high-pressure liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). The equipment required for the instrumental analysis method is expensive and requires a spe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/535G01N33/577
Inventor 李江綦艳李晓明陈满英黎丁滔黄志刚田秀梅
Owner GUANGDONG TESTING INST OF PROD QUALITY SUPERVISION
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