Method for simultaneously determining bacitracin B2 and bacitracin B3 contents in pig tissue

A technology of bacitracin and tissue, applied in the field of simultaneous determination of bacitracin B2 and B3 content in porcine tissue

Pending Publication Date: 2022-07-15
NINGBO ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] In view of the risk of drug residues caused by the use of bacitracin, a series of bacitracin maximum residue limits (MRL) have been formulated at home and abroad for monitoring. For example, the Chinese standard GB31650-2019 stipulates that the MRL of bacitracin in edible tissues of cattle, pigs and poultry is 500 μg / kg, the bacitracin residue markers specified in it are three components of bacitracin A, B, and C, but currently only bacitracin A is measured in the current national standards, or some documents have tested bacitracin B At present, there is no residual analysis method to accurately quantify the bacitracin B2 and B3 respectively. Since there are certain toxicological differences between bacitracin B2 and B3, quantitative analysis of them will be more beneficial to the risk of bacitracin residues. scientific assessment

Method used

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  • Method for simultaneously determining bacitracin B2 and bacitracin B3 contents in pig tissue
  • Method for simultaneously determining bacitracin B2 and bacitracin B3 contents in pig tissue
  • Method for simultaneously determining bacitracin B2 and bacitracin B3 contents in pig tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Pretreatment method

[0034] a. Accurately weigh 2.00 g of the homogenized pork sample, add 10 mL of extraction solvent (1% methanol formic acid), disperse and extract by high-speed homogenization at 10000 r / min for 1 min, centrifuge at 9500 r / min for 2 min, and layer to obtain supernatant A, The remaining residue was repeatedly extracted according to the above steps once to obtain supernatant B. Combine the two supernatants to obtain solution C;

[0035] b. Add 10 mL of n-hexane to the solution C obtained in step a, shake for 1 min, centrifuge at 9500 r / min for 2 min, separate layers, and discard the upper layer to obtain solution D;

[0036] c. Take 2 mL of the solution D obtained in step b with 100 mg of N-propylethylenediamine (PSA), 100 mg of octadecyl-bonded silica gel (C 18 ) mixing, after shaking and purifying for 1min, centrifuging at 9500r / min for 2min, and layering to obtain solution E;

[0037]d. Take 1.0 mL of the solution E obtained in step c and pa...

Embodiment 2

[0058] (1) Pretreatment method

[0059] a. Accurately weigh 2.00 g of the homogenized pig liver sample, add 10 mL of extraction solvent (1% methanol formic acid), disperse and extract by high-speed homogenization at 10000 r / min for 1 min, centrifuge at 9500 r / min for 2 min, and layer to obtain supernatant A , and the remaining residue was repeatedly extracted according to the above steps once to obtain supernatant B. Combine the two supernatants to obtain solution C;

[0060] b. Add 10 mL of n-hexane to the solution C obtained in step a, shake for 1 min, centrifuge at 9500 r / min for 2 min, separate layers, and discard the upper layer to obtain solution D;

[0061] c. Take 2 mL of the solution D obtained in step b with 100 mg of N-propylethylenediamine (PSA), 100 mg of octadecyl-bonded silica gel (C 18 ) mixing, after shaking and purifying for 1min, centrifuging at 9500r / min for 2min, and layering to obtain solution E;

[0062] d. Take 1.0 mL of the solution E obtained in st...

Embodiment 3

[0075] (1) Pretreatment method

[0076] a. Accurately weigh 2.00 g of the homogenized pig kidney sample, add 10 mL of extraction solvent (1% methanol formic acid), disperse and extract by high-speed homogenization at 10000 r / min for 1 min, centrifuge at 9500 r / min for 2 min, and layer to obtain supernatant A , and the remaining residue was repeatedly extracted according to the above steps once to obtain supernatant B. Combine the two supernatants to obtain solution C;

[0077] b. Add 10 mL of n-hexane to the solution C obtained in step a, shake for 1 min, centrifuge at 9500 r / min for 2 min, separate layers, and discard the upper layer to obtain solution D;

[0078] c. Take 2 mL of the solution D obtained in step b with 100 mg of N-propylethylenediamine (PSA), 100 mg of octadecyl-bonded silica gel (C 18 ) mixing, after shaking and purifying for 1min, centrifuging at 9500r / min for 2min, and layering to obtain solution E;

[0079] d. Take 1.0 mL of the solution E obtained in s...

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Abstract

The invention discloses a method for simultaneously determining the contents of bacitracin B2 and bacitracin B3 in pig tissues, which adopts a liquid chromatography-tandem mass spectrometry method for determination, and the chromatographic conditions are as follows: a mobile phase is chromatographically pure acetonitrile and a 0.1% formic acid solution; the mobile phase ratio is as follows: the mobile phase ratio is 20% chromatographic pure acetonitrile in 0-0.2 min, the mobile phase ratio is changed from 20% chromatographic pure acetonitrile in 0.2-6.0 min to 30% chromatographic pure acetonitrile, the mobile phase ratio is changed from 30% chromatographic pure acetonitrile in 6.0-6.1 min to 80% chromatographic pure acetonitrile, the mobile phase ratio is changed from 80% chromatographic pure acetonitrile in 6.1-7.0 min, the mobile phase ratio is changed from 80% chromatographic pure acetonitrile in 7.0-7.1 min to 20% chromatographic pure acetonitrile, and the mobile phase ratio is changed from 7.1-9.0 min to 20% chromatographic pure acetonitrile; the flow velocity of the mobile phase is 0.300 mL / min; the sample size is 10.0 microliters.

Description

technical field [0001] The invention relates to the technical field of bacitracin detection, and more particularly to a method for simultaneously measuring the contents of bacitracin B2 and B3 in pig tissues. Background technique [0002] Bacitracin is an important polypeptide antibiotic, mainly produced by Bacillus licheniformis and Bacillus subtilis. All are effective, so it is widely used in the field of animal husbandry. Bacitracin is a multi-component compound, containing A, B, C, D, E and other components. At present, bacitracin A is the main active ingredient in commercial bacitracin preparations, followed by bacitracin B, and bacitracin B includes components such as B1, B2, and B3. [0003] In response to the risk of drug residues caused by the use of bacitracin, a series of maximum residue limits (MRLs) for bacitracin have been formulated at home and abroad for monitoring. For example, the Chinese standard GB31650-2019 stipulates that the MRL of bacitracin in edib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/30G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/30G01N30/32G01N30/34G01N30/72G01N2030/324
Inventor 王全胜张亮凌淑萍吴银良
Owner NINGBO ACAD OF AGRI SCI
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