Method for simultaneously detecting ochratoxin A and zearalenone

A technology for zearalenone and ochratoxin, applied in the field of analysis and detection, can solve problems such as poor accuracy, and achieve the effects of improving accuracy and sensitivity, improving recovery rate, and high sensitivity and accuracy

Inactive Publication Date: 2017-12-12
INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention solves the problem of poor accuracy of the existing methods for simultaneously measuring ochratoxin A and zearalenone in grains, and further pr

Method used

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  • Method for simultaneously detecting ochratoxin A and zearalenone
  • Method for simultaneously detecting ochratoxin A and zearalenone
  • Method for simultaneously detecting ochratoxin A and zearalenone

Examples

Experimental program
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Embodiment 1

[0055] Weigh 0.01mg of ochratoxin A and 0.1mg of zearalenone respectively, mix them, dissolve them with mobile phase and dilute to 1mL, pass through a 0.45μm organic filter membrane to obtain the standard test solution;

[0056] Adopt high-performance liquid chromatograph to detect standard substance for testing solution, the HPLC spectrogram that obtains is as follows figure 1 as shown, figure 1 Among them, the peak at the retention time of 8.817min is the absorption peak of ochratoxin A, and the peak at the retention time of 10.037min is the absorption peak of zearalenone; wherein the chromatographic conditions are as follows:

[0057] Column: C 18 Column, 250mm×4.6mm, 5μm;

[0058] Mobile phase: acetonitrile: water: 2v% glacial acetic acid with a volume ratio of 49.5:49.5:1;

[0059] Flow rate: 1.0mL / min;

[0060] Detection wavelength: excitation wavelength 333nm, emission wavelength 477nm;

[0061] Injection volume: 10μL;

[0062] Column temperature: 25°C.

[0063] ...

Embodiment 2

[0082] The method for simultaneously detecting ochratoxin A and zearalenone provided in this embodiment comprises the following steps:

[0083] S1. Sample pretreatment:

[0084] Weigh 25.00g of wheat powder, add 150mL of acetonitrile and water mixed solvent with a volume ratio of 8:1, shake and extract for 40min, collect the extract, filter, absorb 10mL of the filtrate, add to 50mL of water to dilute and mix;

[0085] Take 10mL of the diluted solution and transfer it to the immunoaffinity column, adjust the pressure so that the solution passes through the immunoaffinity column at a flow rate of 1-2 drops / s, until some air enters the column, then rinse the column with water twice, wait until the water flow After drying, use 2.0mL methanol for one elution, and control its flow rate to 3 drops / 4s. After the second eluent was drained, combine the two eluents, blow dry under nitrogen at 50°C, cool the residue to room temperature, dissolve it with mobile phase and dilute to 1 mL, p...

Embodiment 3

[0095]The method for simultaneously detecting ochratoxin A and zearalenone provided in this embodiment comprises the following steps:

[0096] S1. Sample pretreatment:

[0097] Weigh 25.00 g of rice powder, add 125 mL of acetonitrile and water mixed solvent with a volume ratio of 9.5:1, shake and extract for 30 minutes, collect the extract, filter, absorb 10 mL of the filtrate, add to 30 mL of water to dilute and mix;

[0098] Take 10mL of the diluted solution and transfer it to the immunoaffinity column, adjust the pressure so that the solution passes through the immunoaffinity column at a flow rate of 1-2 drops / s, until some air enters the column, then rinse the column with water twice, wait until the water flow After drying, use 1.5mL methanol for one elution, control its flow rate to 1 drop / s, and then use 2.0mL methanol for a second elution, control its flow rate to 3 drops / 2s, wait for After the second eluent was drained, combine the two eluents, blow dry under nitrogen...

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Abstract

The invention provides a method for simultaneously detecting ochratoxin A and zearalenone. The method comprises the steps of performing extraction on cereals and/or cereal products by using acetonitrile-water mixed solvent, collecting extracting solution, filtering, diluting filtrate and then transferring to an immunoaffinity column, eluting with methyl alcohol for at least two times, combining eluent and drying by blowing, dissolving and filtering residue to acquire test liquid; and detecting the test liquid by high performance liquid chromatography. The detection method provided by the invention has detection limits of 0.7ng/ml and 4ng/ml for the ochratoxin A and the zearalenone, separately achieves ochratoxin A recovery rates of 98.4%, 99.6% and 99.3% when standard addition levels are 2.5, 5 and 10 microgram/kg, and separately achieves zearalenone recovery rates of 95.17%, 95.23% and 98.62% when standard addition levels are 30, 60 and 120 microgram/kg, which are far higher than the prior art. Therefore, the method provided by the invention is high in sensitivity and accuracy, and can simultaneously detect the ochratoxin A and the zearalenone.

Description

technical field [0001] The invention belongs to the technical field of analysis and detection, and in particular relates to a method capable of simultaneously detecting ochratoxin A and zearalenone in grain and its products. Background technique [0002] Mycotoxins are toxic secondary metabolites produced by fungal growth. In the process of food or feed production, temperature and humidity provide suitable conditions for the growth and reproduction of fungi, thereby causing mycotoxin pollution. According to the report of the World Food and Agriculture Organization, about 25% of the crops in the world are polluted by fungi and their toxins every year, and about 2% of the crops lose their nutritional and economic value due to serious pollution. [0003] Mycotoxins are harmful to human and animal health, especially ochratoxin and zearalenone, which are not only widely distributed, but also more harmful to human health. Ochratoxin is a group of secondary metabolites produced by...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/14G01N33/53
CPCG01N30/02G01N30/14G01N33/53
Inventor 杜晶晶丁勇宝李慧娟王克超宋晓东常建军
Owner INNER MONGOLIA MENGNIU DAIRY IND (GRP) CO LTD
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