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A technology of aflatoxin and determination method, which is applied in the field of determination of aflatoxin and achieves the effects of high safety, reduced pollution and less pollution
Active Publication Date: 2014-07-30
南通市产品质量监督检验所
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Problems solved by technology
However, the current high-performance liquid chromatography analysis methods for aflatoxin all use immunoaffinity columns to enrich the toxin, and the shortcomings of the antibodies mentioned in the enzyme-linked immunoassay method are still unavoidable. Therefore, a new aflatoxin-rich method is studied. Set method to get rid of the shortcomings of antibodies
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Embodiment 1
[0027] (1) Sample extraction: Weigh 10.0346g of edible vegetable oil, add acetonitrile-water extract and stir for 30 minutes, where the volume ratio of acetonitrile and water is 84:16, and then filter with qualitative filter paper.
[0028] (2) Determination of standard solution: take aflatoxin B 1 , G 1 Standard stock solution 10μg / mL, dilute to 5mL with acetonitrile, mix well, and store at 2-8°C for three months; when using, acetonitrile is formulated into 0.025μg / mL, 0.05μg / mL, 0.1μg / mL, 0.15μg / mL , 0.2μg / mL, 0.25μg / mL series standard solutions, draw 200μL of each standard series solution, blow dry under nitrogen in a water bath at 60°C, add 200mL n-hexane and 100mL trichloroacetic acid and mix, after 30s, derivatize in a 40°C oven for 15min , put it in a desiccator for 1min at room temperature, dissolve it with 200uL of water-acetonitrile, mix it evenly, and put it into an automatic sampling bottle. figure 2 , the aflatoxin G1 curve is shown in image 3 .
[0029] (3)...
Embodiment 2
[0038] (1) Sample extraction: Weigh 10.1679g corn flour, add acetonitrile-water extract and stir for 30 minutes, where the volume ratio of acetonitrile and water is 84:16, and then filter with qualitative filter paper.
[0039] (2) Sample determination: use high performance liquid chromatography to detect aflatoxin in the sample
[0040] Chromatographic conditions: column: Agilent SB C-18 (25cm×4.6mm); mobile phase: 85% acetonitrile and 15% water by volume; column temperature: 30°C, injection volume: 25mL; flow rate: 1.0mL / min.
[0041] Take 8mL of extraction filtrate to Mycosep TM 228 multifunctional purification column, transfer 2 mL of purification solution into the derivatization bottle in the collection tank of the purification column, after nitrogen blowing at 60 °C, add 200 mL of n-hexane and 100 mL of trichloroacetic acid and mix well, after 30 s, derivatize in a 40 °C oven for 15 min, at room temperature After drying for 1min, dissolve it with 200uL water-acetonitr...
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Abstract
The invention discloses a method for determination of aflatoxin. The invention is innovative by comprising the following steps: 1) weighing a sample, adding an acetonitrile-water extract, stirring for 30 minutes, and then filtering by using qualitative paper, wherein the acetonitrile and water are in the volume ratio of 84:16; 2) determining the sample under the following conditions: chromatographic conditions of a liquid chromatographic column AgilentSBC-18 (25 cm*4.6 mm), a mobile phase of 85 v% of acetonitrile and 15 v% of water, column temperature at 30 DEG C, sample size of 25 mL and flow rate of 1.0 mL / min; 8 mL of extraction filtering liquid to a multifunctional purification column, transferring 2 mL of purification liquid from a collection pool of a purification column into a derivative bottle, blowing nitrogen at 60 DEG C, adding 200 mL of normal hexane and 100 mL of trichloroacetic acid, mixing uniformly for 30 s, derivatizing for 15 min at a 40 DEG C baking oven, drying at room temperature, dissolving with 200 uL of water-acetonitrile solution, mixing uniformly, and sending into an automatic sample introduction bottle, and determining. The detection method provided by the invention is simple and fast in operation and accurate and reliable in results, has little pollution, high security, reduced pollution to operation personnel and the environment, and low detection cost, and is easy for popularization.
Description
technical field [0001] The invention relates to a method for determining aflatoxin, which belongs to the technical field of food safety inspection. Background technique [0002] Aflatoxins (AF for short) are mainly metabolites of Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are mainly found in soil, animals and plants, various nuts, especially peanuts and walnuts. Aflatoxins are also frequently found in soybeans, rice, corn, macaroni, condiments, milk, dairy products, cooking oils, and other products. Twelve species have been isolated and identified, including B 1 , B 2 , G 1 , G2, M1, M2, P1, Q, H1, GM, B2a, and toxin. [0003] There are three main methods for the analysis of aflatoxins: thin layer chromatography, high performance liquid chromatography and enzyme-linked immunosorbent assay. Thin-layer chromatography is still used at home and abroad because of its simple equipment and easy popularization. However, due to the cumbersome sample pretreatment...
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