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
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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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