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A technology of ochratoxin and detection method, which is applied in the field of detection of ochratoxin A in grains, can solve the problem of high cost and achieve the effect of an accurate analysis method
Inactive Publication Date: 2017-05-10
JIANGSU ACAD OF AGRI SCI
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The pretreatment method for the detection of ochratoxin A in grains at home and abroad is mainly the immunoaffinity column method, because of its high selectivity, strong specificity, high sample recovery rate, and high accuracy, but the disadvantage is that the cost is too high
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Embodiment 1
[0046] Embodiment 1 establishes standard curve
[0047] (1) According to the method of GB / T 25220-2010, the wheat samples and corn samples purchased from the market were tested, and the wheat and corn samples not detected by OTA (such as figure 1 shown, where figure 1 A is a blank sample of wheat, figure 1 B is a corn blank sample) as a blank sample;
[0048] Mix acetonitrile and water at a volume ratio of 4:1 to prepare an acetonitrile solution, add the blank sample powder (ground through a 0.5mm sieve) respectively at a volume to mass ratio (mL / g) of 5:1, shake on a shaker at 180rpm for 30min , centrifuge at 2500rpm for 5min to collect the supernatant; pass 5mL of acetonitrile solution through an amino solid-phase extraction column (500ng, 6mL. Anpu Company), discard the eluate; take 5mL of the supernatant and pass through an amino solid-phase extraction column, discard the column liquid ; Then take 5mL of acetonitrile solution and rinse twice through the column, and disc...
Embodiment 2
[0069] Example 2 sample detection
[0070] 1. Preparation of sample purification solution
[0071] Take the wheat and corn samples collected in the market, and analyze and test according to the following operation process respectively:
[0072] (1) Take wheat and corn samples, crush them and sieve them (the samples are crushed by a pulverizer until they can completely pass through a 0.5mm sieve), and store them in a refrigerator at 4°C for later use;
[0073] (2) After taking the sample out and returning it to room temperature, take 5g of the sample powder and place it in a 150mL Erlenmeyer flask, add 25mL of acetonitrile-water 80:20 (v / v) solution, shake at 180r / min for 30min, then centrifuge at 2500rpm for 5min , take the supernatant;
[0074] (3) Pass 5 mL of acetonitrile-water 80:20 (v / v) through the amino solid-phase extraction column, and discard the eluate;
[0075] (4) Take 5 mL of the supernatant and pass it through the amino solid-phase extraction column, discard ...
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Abstract
The invention discloses a detection method of ochratoxin A in cereals. The detection method comprises steps as follows: a cereal sample is dissolved in an acetonitrile solution, a mixed solution is centrifuged, a supernatant is collected and injected to an amine solid phase extraction column for elution, an eluent is collected and blow-dried, residues are obtained and re-dissolved in a methanol solution, filtering is performed, filtrate is a sample purification liquid, the sample purification liquid is subjected to HPLC-MS-MS (high performance liquid chromatography-tandem mass spectrometry) detection, a detection result is substituted into a curve of a formula 1 for comparison, and the content of ochratoxin A in the sample is obtained. The detection method solves the problem of high cost caused by adoption of an immunoaffinity column for detection of ochratoxin A in the prior art, has the characteristics of being simple, fast, sensitive and accurate, and is applicable to detection of massive samples, and an accurate analysis method can be provided for quality security risk assessment of agricultural products.
Description
technical field [0001] The invention relates to the technical field of agricultural product quality and safety detection, in particular to a method for detecting ochratoxin A in grains. Background technique [0002] Ochratoxins are a group of toxic metabolites with similar structures produced by toxin-producing strains such as Aspergillus and Penicillium. Ochratoxins include seven structurally similar compounds, among which ochratoxin A (ochratoxins A, OTA) is the most toxic and widely distributed. Ochratoxin A is soluble in polar organic solvents and dilute sodium bicarbonate solution, slightly soluble in water, the chemical name of ochratoxin A is 7-(L-β-phenylpropylamino-carbonyl)-carboxy -5-Chloro-8-hydroxy-3,4-dihydro-3R-methylisoxanone (coumarin), the molecular formula is C 2 Oh 18 ClNO 6 . Animal toxicity tests show that ochratoxin A has immunosuppressive toxicity, neurotoxicity, teratogenicity and carcinogenicity. In 1993, the International Center for Cancer Re...
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