A method for extracting and purifying residual neomycin in aquatic products

A technology for neomycin and aquatic products, applied in the field of extraction and purification of neomycin residues in aquatic products, can solve problems such as complex protein matrix, achieve improved detection sensitivity, good specificity, and reduce batch-to-batch differences Effect

Active Publication Date: 2019-05-03
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional extraction and purification method of neomycin is mainly solid-phase extraction. Although solid-phase extraction has good general adaptability, due to the complex problem of protein matrix in aquatic products, there is no suitable solid-phase extraction method applied so far. Study on the extraction and purification methods of neomycin in aquatic products

Method used

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  • A method for extracting and purifying residual neomycin in aquatic products
  • A method for extracting and purifying residual neomycin in aquatic products
  • A method for extracting and purifying residual neomycin in aquatic products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1) neomycin extraction:

[0041] Cut the edible part of the aquatic product into small pieces no larger than 0.5cm×0.5cm×0.5cm, mix well, and fully homogenate; then weigh 1.0g and place it in a centrifuge tube, add 10mL sample extract, shake for 5min, The tissue was dispersed for 1 min, and then centrifuged at 10,000 r / min at 4°C for 10 min; among them, the supernatant was transferred to another centrifuge tube, and the pH was adjusted to 7.5 with 6mol / L NaOH; 10 mL of deionized water was added to the residue, and the first After treatment by the extraction method, combine the supernatants obtained twice, add 10 mL of n-hexane and let stand for 10 min, remove the n-hexane layer, add 15 mL of deionized water to the supernatant to obtain an extract.

[0042] Wherein, the preparation method of the sulfosalicylic acid solution is as follows: Weigh EDTA-Na2 in a beaker, add deionized water at a solid-to-liquid ratio of 1.5g / 800mL, and ultrasonically treat at 35°C for 20min; ...

Embodiment 2

[0074] Take the edible part of turbot purchased from Liqun Supermarket (Qingdao), cut into small pieces no larger than 0.5cm×0.5cm×0.5cm, mix well, fully homogenate, take 1.00g (accurate to 0.01g) in a 50mL centrifuge tube. Accurately weigh 1.0g of fish meat (accurate to 0.01g), place it in a 50mL polypropylene centrifuge tube, add 10mL of 2% sulfosalicylic acid, shake for 5min, disperse at high speed for 1min, and then centrifuge at 10000r / min at 4 degrees After 10 minutes, the supernatant was transferred to another 50mL centrifuge tube, and the pH was adjusted to about 7.5 with 6mol / L NaOH. Add 10 mL of 2% sulfosalicylic acid to the residue, shake for 5 min, disperse at high speed for 1 min, centrifuge at 10,000 r / min at 4 degrees for 10 min, combine the supernatants, add 10 mL of n-hexane and let stand for 10 min, remove the n-hexane layer, and Add 15mL deionized water to the clear solution to dilute.

[0075] The sample extract was purified by an enzyme-linked immunoaffi...

Embodiment 3

[0077] Take the edible part of turbot purchased from Liqun Supermarket (Qingdao), cut into small pieces no larger than 0.5cm×0.5cm×0.5cm, mix well, fully homogenate, take 1.00g (accurate to 0.01g) in a 50mL centrifuge tube. Accurately weigh 1.0g of fish meat (accurate to 0.01g), place it in a 50mL polypropylene centrifuge tube, add 10mL of 2% sulfosalicylic acid, shake for 5min, disperse at high speed for 1min, and then centrifuge at 10000r / min at 4 degrees After 10 minutes, the supernatant was transferred to another 50mL centrifuge tube, and the pH was adjusted to about 7.5 with 6mol / L NaOH. Add 10 mL of deionized water to the residue, shake for 5 min, disperse at high speed for 1 min, centrifuge at 10,000 r / min at 4°C for 10 min, combine the obtained supernatants, add 10 mL of n-hexane and let stand for 10 min, remove the n-hexane layer.

[0078]The sample extract was purified by an enzyme-linked immunoaffinity column, and the sample extract was passed through the enzyme-li...

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Abstract

The invention relates to the technical field of pesticide residue detection, and discloses a method for extracting and purifying neomycin remaining in an aquatic product. The method includes the steps that 1, neomycin is extracted, wherein an aquatic product sample is diced, the dices are evenly mixed, fully homogenized and then put into a centrifuge tube, sample extraction liquor is added for extraction, and extract liquor is obtained; 2, neomycin is purified, wherein an ELISA affinity column is placed at the room temperature in advance to be restored to the normal temperature, and balanced activation is carried out with a phosphate buffer solution; the obtained extract liquor passes through the ELISA affinity column, then the ELISA affinity column is leached with leaching liquor, leacheate is discarded, the leaching liquor is fully removed, elution is carried out with an organic reagent and inorganic acid eluent in sequence, and eluate is collected. The method is high in extraction efficiency, high in recovery rate, good in specificity, easy to implement and low in cost.

Description

technical field [0001] The invention relates to the technical field of drug residue detection, in particular to a method for extracting and purifying residual neomycin in aquatic products. Background technique [0002] Neomycin is a representative of aminoglycoside antibiotics and one of the veterinary drugs commonly used in my country's agriculture, animal husbandry and aquaculture. It is often used to prevent and control fish diseases, and it is also added to feed to promote growth and development. However, it can form residues in animal tissues and endanger human health through the food chain. Therefore, the harm of antibiotics such as neomycin residues in aquatic products has been paid more and more attention all over the world. The European Union stipulates that aminoglycosides (streptomycin, neomycin) should not be used as feed additives. The European Union, the United States, Japan and my country stipulate that the maximum residue limit (MRLs) of neomycin in animal ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N1/34
CPCG01N1/28G01N1/34G01N1/405
Inventor 隋建新郑洪伟曹立民林洪郑琦崔梦琪屈雪丽曲欣陈静张鑫磊
Owner OCEAN UNIV OF CHINA
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