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Method for detecting erythromycin residue in edible animal tissues

A technology of erythromycin and analysis method, applied in the field of veterinary drug residue analysis, to achieve the effects of high ultraviolet signal response, quality improvement and sensitivity improvement

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

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Problems solved by technology

It can be seen that there are many reports on the detection of erythromycin residues, especially the residues of macrolides, by liquid chromatography tandem mass spectrometry. The above three national standards are all confirmation methods, and there are relatively few screening methods. , poultry muscle, fat, liver, kidney and milk, eggs, erythromycin residue detection HPLC screening method has not been reported

Method used

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  • Method for detecting erythromycin residue in edible animal tissues
  • Method for detecting erythromycin residue in edible animal tissues
  • Method for detecting erythromycin residue in edible animal tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Embodiment 1: Derivatization reaction of hydroxyl compound and 9-fluorenylmethoxybenzoyl chloride

[0094] (1) Derivatization reaction of erythromycin with 9-fluorenylmethoxybenzoyl chloride

[0095] Accurately weigh 10.0 mg of erythromycin, dissolve it in methanol, dilute to 10 mL, and prepare a 1 mg / mL standard stock solution. Accurately pipette an appropriate amount of erythromycin standard stock solution, and dilute it successively with acetonitrile to a concentration of 10 μg / L, 50 μg / L, 100 μg / L, 250 μg / L, 500 μg / L, 1000 μg / L, 2000 μg / L, 4000 μg / L 600μL standard solution, add 300μL 0.1mol / L KH2PO4 solution (pH7.5) and 100μL 0.5mg / mL FMOC-CL, mix well and react at 45℃ for 1h. After the reaction, cool to room temperature, and after standing at room temperature for 20 minutes, take 20 μL for high-performance liquid chromatography determination.

[0096] (2) HPLC detection of derivative products

[0097] Determination is carried out according to the following HPLC ...

Embodiment 2

[0102] Embodiment 2: sample measurement, analysis

[0103] (1) Processing of samples

[0104] The muscle, liver, kidney, fat, milk, and eggs of cattle, sheep, pigs, and poultry were selected as experimental samples, and erythromycin was added and recovered (refer to the literature for the test method:). Add different concentrations of erythromycin standard solutions to blank tissue samples so that the erythromycin concentration in the sample is 4, 200 and 400 μg / kg, respectively (the erythromycin concentration in milk is 4, 40 and 80 μg / kg) , according to the sample processing method, extraction, purification, derivatization, and then perform HPLC analysis (see the literature for the test method:). Each concentration was repeated 5 times, and 5 separate experiments were carried out, and the recovery rate and the inter-day (or inter-batch) coefficient of variation were calculated by the external standard method.

[0105] 1) Sample extraction: Accurately weigh 5g samples (accu...

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Abstract

The invention discloses a method for detecting erythromycin residue in edible animal tissues, belongs to the technical field of veterinary drug residue detection, and particularly relates to a high performance liquid chromatography pre-column derivatization reaction analysis method for the erythromycin residue in the animal tissues. Hydroxyl groups contained in erythromycin drug react with 9-fluorenyl methoxyl benzoyl chloride, and the product has a relatively high ultraviolet signal response. In a slightly-alkaline monopotassium phosphate buffer solution, a derivatization reagent 9-fluorenyl methoxyl benzoyl chloride is used to derive erythromycin for one hour in a water bath at 45 DEG C; after the reaction ends, the reaction product is cooled to the room temperature; HPLC (High Performance Liquid Chromatography) analysis detection is performed on the reaction product, so as to obtain a relatively-stable derivative; ultraviolet spectrum scanning is performed on a standard erythromycin derivative solution through a diode array detector; maximum absorption exists at 264 nm. The method has the advantages as follows: the sensitivity is 4 [mu]g / kg; the linear response is shown to the peak area within the range of 0.1-2 [mug] / mL; the result reproducibility is good; the within-run and between-run variable coefficient CV is smaller than or equal to 15%; all indexes can meet the requirements of the erythromycin residue analysis; the sensitivity is improved greatly.

Description

technical field [0001] The invention belongs to the technical field of veterinary drug residue analysis, and relates to a method for detecting erythromycin residue in edible tissues in animal tissues. Background technique [0002] Erythromycin (Erythromycin, EM) has another name called Weimycin, molecular formula C 37 h 67 NO 13 , the molecular weight is 733.93, the melting point is 138-140°C, and it is weakly alkaline. Erythromycin is white or off-white crystal or powder; odorless; bitter; slightly hygroscopic. Erythromycin is easily soluble in methanol, ethanol or acetone, and slightly soluble in water. The pH value of 0.066% aqueous solution should be 8.0-10.5. The dry state of erythromycin is relatively stable in neutral and weakly alkaline solutions, but unstable in acidic conditions, and will be destroyed immediately when the pH value is lower than 4. Its structural formula is as follows: [0003] [0004] Erythromycin is a special macrolide antibiotic for an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 袁宗辉陶燕飞陈冬梅潘源虎王玉莲黄玲利王旭刘振利谢书宇彭大鹏戴梦红郝海红程古月
Owner HUAZHONG AGRI UNIV
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