ZHAO Jie,LI Qiaoning,LIU Ailing,et al.Establishment of high performance liquid chromatography method for measuring cefquinome sulfate residues in eggs[J].Journal of Nanjing Agricultural University,2017,(4):703-709.[doi:10.7685/jnau.201612026]





Establishment of high performance liquid chromatography method for measuring cefquinome sulfate residues in eggs
赵婕1 李巧宁1 刘爱玲2 孟小宾2 鲍恩东12
1. 南京农业大学动物医学院, 江苏 南京 210095;
2. 瑞普(天津)生物药业有限公司, 天津 300308
ZHAO Jie1 LI Qiaoning1 LIU Ailing2 MENG Xiaobin2 BAO Endong12
1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2. Ringpu(Tianjin)Bio-Pharmacy Co.Ltd., Tianjin 300308, China
cefquinome sulfateeggHPLCassey of residue
[目的]为了解硫酸头孢喹肟在鸡蛋中的残留,以确定蛋鸡用药的合理休药期,建立了硫酸头孢喹肟在鸡蛋内的高效液相色谱(HPLC)残留检测方法。[方法]取空白鸡蛋样品,按质量体积比1∶19的比例添加相应浓度的硫酸头孢喹肟对照品溶液制备生物样品,用酸性乙腈提取药物,正己烷分离脂肪,C18固相萃取小柱净化洗脱后,洗脱液经40 ℃水浴N2吹干浓缩后复溶,在紫外吸收波长270 nm下进行HPLC检测。[结果]硫酸头孢喹肟在0.02~2.00 μg·;mL-1 浓度范围内线性关系良好,回归方程为:y=164.95x+0.693(R2=0.999 8);方法的最低检测限0.01 μg·;mL-1,定量限0.02 μg·;mL-1;药物在生物样品中的低、中、高浓度分别为0.01、0.10和1.00 μg·;mL-1 时,鸡蛋中硫酸头孢喹肟的回收率为85.6%~90.4%,变异系数为6.59%~10.47%,均在规定的药物残留检测添加回收率和变异系数范围内。此外,该检测方法的特异性好、准确度高,24 h内生物样品在室温、4 ℃(冷藏)和-20 ℃(冷冻)条件下稳定性均良好,且临床常用抗菌药物不存在干扰。[结论]经方法学验证,本方法适用于鸡蛋中硫酸头孢喹肟含量的检测,可应用于临床的生物样品检测。
[Objectives]To understand the elimination distribution of residue and the determination of the withdrawal time of cefquinome sulfate in eggs,the detection method of high performance liquid chromatography(HPLC)was established. [Methods]The samples were made by adding cefquinome sulfate into blank eggs according to the quality and volume proportion at 1∶19. After being mixed totally,the cefquinome sulfate was extracted from eggs by acidic acetonitrile solution(adding 50 μL formic acid into 100 mL solution contented with 10 mL acetonitrile),mixing up totally for three times. After being degreased in n-hexane for 30 minutes,the extract solution was cleaned up on C18solid-phase extraction cartridges. The eluent was collected and dried under nitrogen evaporator in 40 ℃ water,and the residues were dissolved in mobile phase. The analyte was detected by high performance liquid chromatography(HPLC),under UV absorptive spectroscopy detector at 270 nm,after being filtered by microporous membrane. [Results]The testing data showed that the calibration curve was linearity at a range of 0.02-2.00 μg·mL-1,with a correlation coefficient(R2)of 0.999 8,which showed good linear relationship,the linear regression equation:y=164.95x+0.693. By comparing the S/N value,the lowest limit of detection(LOD)was 0.01 μg·mL-1,and the lowest limit of quantitation(LOQ)was 0.02 μg·mL-1 for cefquinome sulfate in the tested eggs,respectively. With the concentrations of cefquinome sulfate at 0.01,0.10 and 1.00 μg·mL-1,the average recoveries were ranged from 85.6% to 90.4%,and the coefficient of variances were ranged from 6.59% to 10.47%,which were all within the prescribed scope. Meanwhile,this method was a specific,sensitive and rapid method for detecting the cefquinome sulfate residue in eggs. The results of stability test showed that,the samples under room temperature,4 ℃(cold storage)and -20 ℃(frozen)were stable within 24 hours,and the interference of clinical commonly used antimicrobial agents was not affected,which could meet the requirement of residue analysis. [Conclusions]With comprehensive analysis on all the testing data,the HPLC method established in this paper is conformed to the requirements of the adaptability and accuracy of residue detection. Meanwhile,this method can also be used in the detection of the cefquinome sulfate in eggs.


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更新日期/Last Update: 2017-07-07