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 recovery rate fluctuation, long operation time, interference, etc., and achieve rapid detection, simple steps and good stability

Active Publication Date: 2017-01-04
GUANGDONG TESTING INST OF PROD QUALITY SUPERVISION
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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
  • Detection method of aflatoxin B1
  • Detection method of aflatoxin B1

Examples

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Embodiment 1

[0048] A detection method for aflatoxin B1, comprising the following steps:

[0049] 1. Pre-processing.

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

[0051] The specific pretreatment method is as follows: Weigh 5.0g of representative samples into a 50mL polystyrene centrifuge tube, add different extraction solutions in different proportions and mix them; vibrate vigorously on an oscillator for 10 minutes at a speed of 150r / min (or vortex for more than 5 minutes); take the liquid and centrifuge it at 4000r / min for 5 minutes (or let it stand for 3 minutes, and then filter it 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 5 s or shake it by hand, and take 50 μL for analysis.

[0052] Second, detection.

[0053] Take the solution to be tested, ...

Embodiment 2

[0120] A detection method for aflatoxin B1, comprising the following steps:

[0121] 1. Pre-processing.

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

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

[0124] Second, detection.

[0125] Detect according to the metho...

Embodiment 3

[0134] A detection method for aflatoxin B1, comprising the following steps:

[0135] 1. Pre-processing.

[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] Concrete pretreatment method is: take by weighing 5.0g representative sample and insert in 100mL tool stopper Erlenmeyer flask, add the methyl alcohol of 25mL60% and mix it as extraction solution; Vigorously vibrate 10 minutes on the shaker, the rotating speed is 150r / min ( Or vortex for more than 5min); take the liquid and centrifuge at 4000r / min for 5min (or let it stand for 3min, and then filter it with quantitative filter paper); take 1mL of supernatant or filtrate, and then add 5mL deionized water as a diluent to obtain the solution to be tested. The pH value of the solution to be tested should be between 6 and 8, which can be adjusted with "NaOH" and "HCL". Shake the solution to be t...

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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 method for detecting aflatoxin B1. Background technique [0002] At present, the detection methods of aflatoxin B1 in soy sauce, vinegar and other foods mainly include chemical analysis, instrumental analysis, immunoassay and biosensor. [0003] Among them, thin-layer chromatography is the most commonly used method in chemical analysis, and thin-layer chromatography (TCL) utilizes the characteristic that certain mycotoxins (such as aflatoxins) can display fluorescence at specific wavelengths. This method is more economical and requires less equipment and inspectors, but it cannot be quantified accurately. [0004] Instrumental analysis methods mainly include high pressure liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). Instrumental analysis method requires expensive instruments and detection costs, requires special p...

Claims

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Application Information

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Patent Type & Authority Applications(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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