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Post-column derivatization preparation method and application of aminoglycosides compound

A post-column derivatization and compound technology, applied in the field of veterinary drug residue analysis, can solve problems that are difficult to control and difficult to find

Inactive Publication Date: 2013-12-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention overcomes the defects that it is difficult to find a suitable mobile phase and difficult to control in the reaction process in the pre-column reaction detection of the liquid phase method, and effectively improves the sensitivity and expansion of high-performance liquid chromatography and fluorescence detection of such drug residues through post-column derivatization. scope of application

Method used

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  • Post-column derivatization preparation method and application of aminoglycosides compound
  • Post-column derivatization preparation method and application of aminoglycosides compound
  • Post-column derivatization preparation method and application of aminoglycosides compound

Examples

Experimental program
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Effect test

Embodiment 1

[0013] In this example, a high performance liquid chromatography (HPLC) separation and detection method for standard samples of aminoglycoside compounds was first established, and then the conditions for derivatization reaction with o-phthalaldehyde were established, and finally the samples were analyzed and detected.

[0014] Specific steps are as follows:

[0015] 1. Purification treatment of aminoglycosides

[0016] Extract with phosphate buffer (weigh 8.50g of sodium chloride, 3.58g of disodium hydrogen phosphate, 3.12g of sodium dihydrogen phosphate, add water to dissolve and dilute to 1000mL) and 10% trichloroacetic acid solution, C 18 SPE column purification, eluting with methanol, collecting the eluate, blowing dry with nitrogen in a water bath at 40°C, dissolving in 1.0 mL of water, transferring to a 1.5 mL polypropylene centrifuge tube, centrifuging at 10,000 r / min for 10 min, and taking the supernatant for high performance liquid chromatography analyze.

[0017] 2...

Embodiment 2

[0042] Embodiment 2: the mensuration of sample

[0043] (1) Extraction of samples

[0044] Select pig kidney, pig muscle, bovine kidney and bovine muscle, chicken muscle and milk as test samples, and carry out single or multiple residue detection of apramycin, kanamycin, neomycin and gentamicin. The concentrations of kanamycin, apramycin and gentamicin added to milk were 20μg / L, 50μg / L, 100μg / L and 200μg / L respectively, and the concentration of neomycin was 100μg / L, 500μg / L, 1000μg / L and 2000μg / L; the concentration of gentamicin added in pig kidney, cow kidney, pig muscle, cow muscle and chicken muscle was 50μg / kg, 100μg / L kg and 200μg / kg three added concentrations, set a blank sample respectively, repeat each concentration 5 times, and carry out 5 separate experiments to calculate the recovery rate and variation coefficient with the commonly used external standard method

[0045] The sample is extracted with 10% trichloroacetic acid solution of phosphate buffer (the formul...

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Abstract

The invention belongs to the technical field of veterinary drug residue analysis, and discloses a detection and analysis method for post-column derivatization reaction of aminoglycosides compound and o-phthalaldehyde. Aminoglycosides compound reacts with o-phthalaldehyde to generate a fluorescent reagent which has unique excitation wavelength and unique emission wavelength. For the optimization selection of the reaction conditions, the result shows that under the alkali condition, aminoglycosides can react easily and quickly with o-phthalaldehyde, so that the reaction is suitable for the post-column derivatization; a derivatization product has the strongest fluorescence intensity at the lambda ex value of 235 nm and lambda em value of 465 nm; the minimum detectable amount is 10 microgram / L with good reproducibility; the variable coefficient of 4 concentrations and 5 independent repetitions is smaller than 15%, so that each index of the method can satisfy the requirements for the residue analysis of aminoglycosides antibiotics, and the sensitivity is greatly increased.

Description

technical field [0001] The invention belongs to the technical field of veterinary drug residue analysis, and in particular relates to a post-column derivatization preparation method and application of aminoglycoside compounds. The method of the invention can be used for residue analysis of aminoglycoside antibiotics in edible animal products. Background technique [0002] Aminoglycosides are a class of antibiotics commonly used in clinical practice. Their basic structure is composed of aglycone and amino sugar molecules connected by oxygen bridges, including gentamicin, streptomycin, dihydrostreptomycin, kanamycin , spectinomycin, neomycin and synthetic tobramycin, amikacin, netilmicin, etc. The mechanism of action of aminoglycoside antibiotics is to affect multiple links in protein biosynthesis to prevent protein synthesis. Aminoglycoside antibiotics are effective against both Gram-positive and Gram-negative bacteria, and have a stronger effect on Gram-negative bacteria. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H15/26C07H19/044C07H1/00G01N30/06
Inventor 袁宗辉陶燕飞陈冬梅潘源虎黄玲利王玉莲王旭刘振利谢叔宇彭大鹏戴梦红郝海红程古月
Owner HUAZHONG AGRI UNIV
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