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Method for detecting toxoflavin in foodstuff based on multifunctional purifying column-high performance liquid chromatography

A technology of high-performance liquid chromatography and multi-functional purification column, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve high detection efficiency, easy promotion and application, and good purification effect

Inactive Publication Date: 2016-09-28
浙江省产品质量安全检测研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, there are few reports on the research on food poisoning flavin detection technology at home and abroad.

Method used

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  • Method for detecting toxoflavin in foodstuff based on multifunctional purifying column-high performance liquid chromatography
  • Method for detecting toxoflavin in foodstuff based on multifunctional purifying column-high performance liquid chromatography
  • Method for detecting toxoflavin in foodstuff based on multifunctional purifying column-high performance liquid chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: Draw the calibration curve of response peak area-toxoflavin concentration.

[0034] (1) Weigh 0.0100g of toxin standard product into a 10mL volumetric flask, dissolve in methanol, set the volume to the mark, and prepare a 1.0mg / mL standard mother solution. Stored in -18°C refrigerator, it can be stored for 3 months. Before use, take it out and let it stand at room temperature, take 200μL of the standard mother solution in a 10mL volumetric flask, dilute to the mark with water, shake well, and prepare a 20.0μg / ml intermediate solution for use. Take 25μL, 50μL, 100μL, 200μL, 500μL, 1.0mL intermediate use solution and 50μL, 100μL standard mother solution in 10mL volume, and dilute water to the mark. Prepare a series of standard solutions with concentrations of 0.05, 0.1, 0.2, 0.4, 1.0, 2.0, 5.0, and 10.0 μg / ml.

[0035] At the same time, this part optimizes the constant volume solution, and compares the influence of different volume fractions of methanol wa...

Embodiment 2

[0037] Embodiment 2: The present embodiment detects toxanthin in fermented cornmeal by the following method:

[0038] (1) Sample extraction conditions: Weigh 20-25 g of sample and mix with 100 mL of extract (80 acetonitrile + 20 water), ultrasonically extract for 30 min, take the extract, and centrifuge at 5000 r / min for 5 min to obtain supernatant.

[0039] (2) Sample purification: Take 10 mL of the supernatant obtained in step 1, and use a multifunctional purification column for purification; after purification, take 5.0 mL of the purification solution and use N 2 Blow dry, then dilute to volume with 1.0mL pure water, vortex to dissolve to obtain supernatant solution.

[0040] (3) Chromatographic analysis conditions: the upper machine liquid obtained in step 2 is subjected to chromatographic analysis through a quaternary low-pressure or binary high-pressure liquid chromatography system equipped with a DAD detector, chromatographic column: ODS reverse-phase chromatographic colu...

Embodiment 3

[0043] Example 3: In this example, in Example 2, a positive fermented cornmeal sample was prepared by adding toxin standard substance into cornmeal at the same time, and the same method was used for detection. Wherein in step 1, this part has carried out the test to extraction reagent, has selected methanol water ( Figure 9 ), 70% ethanol water ( Figure 10 ) and 70% acetonitrile water ( Figure 11 ), to investigate the influence of different organic solvents on the extraction efficiency and the effect of purification and recovery on the multifunctional purification column. Among them, the influence of different organic solvents on the extraction efficiency was obtained by extracting the positive corn fermented noodles, and then taking 3.0ml of the extract solution to be directly dried with N2, dissolved in water and analyzed on the machine, so as to eliminate the influence of recovery rate caused by column purification. In addition, each part of the extract was purified by...

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Abstract

The invention provides a method for detecting a toxoflavin residue in foodstuff based on multifunctional purifying column purification-high performance liquid chromatography. 80mL of acetonitrile and 20mL of water are used for dissolving a foodstuff sample, ultrasonic extraction is carried out, a multifunctional purifying column is used for purifying, 5.0mL of purifying liquid is used, N2 is used for blowing and drying, 1.0mL of water is used for metering the volume, and liquid for testing is obtained. Detection of the toxoflavin residue in the sample is realized, and the method has the advantages of good purifying effects, accurate quantification, and elimination of influences of solvent effect on chromatographic behaviors of toxoflavin. Gradient elution of a mobile phase is carried out by methanol and water, retention of toxoflavin in a reversed phase chromatography column, effective separation between toxoflavin and other interfering substances is realized during sample analysis, so that the purpose of accurate quantification is realized. In the sample purifying process, interferences of lipids, pigments, proteins and other substances are effectively removed from the sample, the purifying effect is good, and the quantitative analysis has good accuracy, high sensitivity, and good result reappearance; and the method is suitable for detection of the toxoflavin residue in a plurality of foodstuffs.

Description

technical field [0001] The invention relates to the technical field of food safety, in particular to a method for detecting food poisoning flavins based on a multifunctional purification column-high performance liquid chromatography. Background technique [0002] The structural formula of toxoflavin (CAS:84-82-2) is: [0003] [0004] It is a bacterial toxin that can cause food poisoning produced by Pseudomonas cocos subsp. Common contaminated foods are mainly fermented cereal products, potato products and white fungus. Food poisoning by Pseudomonas cocos subspecies fermented rice noodles mostly occurs in summer and autumn (mainly from May to August) during the rainy and humid seasons. Improper food storage causes Pseudomonas cocos to multiply and grow rapidly, and food contaminated by it is eaten It can cause poisoning, manifested as symptoms such as epigastric discomfort, nausea, vomiting, dizziness, and general weakness. In severe cases, hepatosplenomegaly, subcutane...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 李红艳翁晨辉
Owner 浙江省产品质量安全检测研究院
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