Study On Detection Method Of Four Veterinary Drug Residues In Eggs By High Performance Liquid Chromatography

This study established residue analytical methods of quinolones, sulfonamides amphenicols, tetracyclines kinds of drugs in eggs. Using the simple traditional liquid-liquid extraction method combined with the solid-phase method to extract and purify eggs samples,Finally using high performance liquid chromatography to analysis. Four kinds of veterinary drug residues in eggs were determined,results are as follows: 1. Research of analysis method on quinolones residue in EggsThe four kinds of quinolones residues determination of eggs by HPLC was established. Samples were extracted by phosphate, wiped off fat by n-hexane and went over PLS cartridge purify for HPLC determination. The results showed that four kinds of quinolones showed good linearity in a range of 50 ng/g~1000 ng/g. The limits of detection were 10 ng/g and the limit of quantification were 50 ng/g. Recoveries ranged from 60.2% to 68.2% for the three different concentrations added(10 ng/g, 200 ng/g, 500 ng/g). Intra-day and Inter-day relative standard deviations were less than 6.6%. 2. Research of analysis method on sulfonamides residue in eggsThe six kinds of sulfonamides residues determination of eggs by HPLC was established. The extraction solvent was ethyl acetate, combined with ultrasonic assisted extraction, then was vacuum evaporated. The residue dissolved in mobile phase, and went over the Oasis MCX solid phase extraction cartridge. 5% ammoniation of methanol was used to purge. Then C18 reverse column was chosen to HPLC analysis. Under the selected chromatographic conditions, six kinds of sulfonamides showed good linearity in a range of 15 ng/g~1000 ng/g. The limits of detection were 5 ng/g and 10 ng/g and the limit of quantification were 15 ng/g and 30 ng/g. Recoveries ranged from 70.1% to 82.5% for the three different concentrations added(10 ng/g, 200 ng/g, 500ng/g).Intra-day and Inter-day relative standard deviations were less than 5.5%. 3. Research of analysis method on amphenicols residue in eggsThe method of HPLC for determination of two kinds of amphenicols residues in eggs was developed. Samples were extracted with acetonitrile- water(85+15).The fat was wiped off by n-hexane and then ethyl acetate was added.The liquid was vacuum evaporated under reduced pressure. The residue was dissolved in acetonitrile-water(5+95), and went over C18 solid phase extraction columns for HPLC determination. The results showed that two kinds of amphenicols showed good linear correlated in the concentration range of 50 ng/g~1000 ng/g. The limits of detection were 10 ng/g and the limit of quantification were 50 ng/g. Two kinds of amphenicols at 10 ng/g, 200 ng/g, 500 ng/g three concentration added the average recovery rate ranged from 60.6% to 71.1%. Intra-day and Inter-day relative standard deviations were less than 6.5%. 4. Research of analysis method on tetracyclines residue in eggsThe method of HPLC for determination of four kinds of tetracyclines residues in eggs was developed.The extraction solvent was Na2EDTA-Mcllvaine solution,purification of PLS solid phase extraction column. Oxalic acid methanol was used to elution. Then the samples were concentrated by 35℃ water bathed and constant volume for HPLC determination. The method of four kinds of tetracyclines showed good linear correlated in the concentration range of 30 ng/g~1000 ng/g.The limits of detection were 10 ng/g, the limits of quantification were 30 ng/g and 50 ng/g. Four kinds of tetracyclines at 10 ng/g, 200 ng/g, 500 ng/g three concentration added the average recovery rate ranged from 61.5% to 74.4%, Intra-day and Inter-day relative standard deviation were less than 5.6%.This paper studied on the analysis method of 4 kinds of veterinary drug residues in eggs.According to the chemical properties of drugs and the characteristics of the egg matrix,optimization the detection of drug residues in samples of various types of pre-treatment methods and chromatographic conditions parameters. Accuracy, sensitivity, specificity, operability, etc were obtained satisfactory results, which can meet 4 drug residues testing requirements in eggs.