Method for detecting erythromycin residue in edible animal tissues
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A technology of erythromycin and analysis method, which is applied in the field of veterinary drug residue analysis to achieve the effects of safeguarding human health, increasing export earnings, and improving quality
Inactive Publication Date: 2015-05-13
HUAZHONG AGRI UNIV
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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
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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 was 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 the addition and recovery experiment of erythromycin was carried out (see the literature for the test method:). Add different concentrations of erythromycin standard solutions to blank tissue samples so that the concentrations of erythromycin in the samples are 4, 200 and 400 μg / kg, respectively (the concentrations of erythromycin in milk are 4, 40 and 80 μg / kg) According to the sample processing method, extraction, purification, derivatization, and then 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: Accur...
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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) is also known as 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 the 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 anima...
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