[1]赵婕,李巧宁,刘爱玲,等.鸡蛋中硫酸头孢喹肟残留的高效液相色谱检测方法的建立[J].南京农业大学学报,2017,(4):703-709.[doi:10.7685/jnau.201612026]
 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]
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鸡蛋中硫酸头孢喹肟残留的高效液相色谱检测方法的建立()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
期数:
2017年4期
页码:
703-709
栏目:
出版日期:
2017-07-07

文章信息/Info

Title:
Establishment of high performance liquid chromatography method for measuring cefquinome sulfate residues in eggs
作者:
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
Author(s):
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
关键词:
硫酸头孢喹肟鸡蛋HPLC残留检测
Keywords:
cefquinome sulfateeggHPLCassey of residue
分类号:
S859.79+6
DOI:
10.7685/jnau.201612026
摘要:
[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。
Abstract:
[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.

参考文献/References:

[1] 荣俊. 国产硫酸头孢喹肟毒理学及对鸡大肠杆菌病疗效学的初步研究[D]. 南京:南京农业大学,2008.Rong J. Primary study on the toxicity and pharmacodynamics of domestic cefquinome slufate[D]. Nanjing:Nanjing Agricultural University,2008(in Chinese with English abstract).
[2] Yehia M,Arafa M,Abbas S,et al. Ratio manipulating spectrophotometry versus chemometry as stability indicating methods for cefquinome sulfate deterrmination[J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2015,153(1):231-240.
[3] Elghobashy R,Bebawy I,Abbas S,et al. SAT-Indicating HPLC and RP-TLC determination of cefpirome sulfate with Kinetic study[J]. Chromatographia,2013,76(17):1141-1151.
[4] Thomas E,Withelm C. Antibacterial activity of cefquinome against equine bacterial pathogens[J]. Veterinary Microbiology,2006,115:140-147.
[5] 刘美瑞,赵地顺,刘宝树,等. 头孢喹肟研究进展[J]. 中国畜牧兽医,2013,40(4):216-220.Liu M R,Zhao D S,Liu B S,et al. The research progress of cefquinome slufate[J]. China Animal Husbandary and Veterinary Medicine,2013,40(4):216-220(in Chinese).
[6] 蒋茂春,陈晓兰,陈毓. 硫酸头孢喹肟在鸡体内的药动学和生物利用度研究[J]. 畜牧与兽医,2014,46(7):73-76.Jiang M C,Chen X L,Chen Y. The research of pharmacokinetics and bioavailability of cefquinome slufate in chicken[J]. Animal Husbandary and Veterinary Medicine,2014,46(7):73-76(in Chinese).
[7] Du X X,Zu S C,Chen F L,et al. Preparation and characterization of cefquinome sulfate microparticles for transdermal delivery by negative-pressure cavitative antisolvent precipitation[J]. Powder Technology,2016,294(6):429-436.
[8] 董艳珍,陈碧华,肖文渊. 一株光合作用细菌分离与初步鉴定[J]. 西昌学院学报(自然科学版),2010,24(3):6-10.Dong Y Z,Chen B H,Xiao W Y. A plant photosynthesis bacteria isolation and preliminary identification[J]. Journal of Xichang College(Natural Science Edition),2010,24(3):6-10(in Chinese).
[9] Zonca A,Gallo M,Locatellli C,et al. Cefquinome sulfate behavior after intramammary administration in healthy and infected cows[J]. Dairy Science,2011,94:3455-3461.
[10] 刘利锋. 硫酸头孢喹肟在猪体内的药动学和残留初步研究[D]. 南京:南京农业大学,2008.Liu L F. Studu on pharmacokinetics and residues of cefquinome in pigs[D]. Nanjing:Nanjing Agricultural University,2008(in Chinese with English abstract).
[11] CVMP(Committee for Veterinary Medicial Products). Cefquinome Summary Report,EMEA/MRL/005/95[R]. London:EMEA,1995.
[12] 李婷婷. 中国与欧盟主要动物源性产品中兽药残留限量差异性研究[D]. 合肥:合肥工业大学,2009.Li T T. Differential study on veterinary drugs residues in the main animal-source products of China and European Union[D]. Hefei:Hefei University of Technology,2009(in Chinese with English abstract).
[13] 杨迪,张春兰,刘彩,等. 磷酸替米考星在肉鸡组织内残留消除规律研究[J]. 南京农业大学学报,2013,36(4):111-115. DOI:10.7685/j.issn.1000-2030.2013.04.018.Yang D,Zhang C L,Liu C,et al. Elimination regularity and residues of tilmicosin phosphate in broiler tissues[J]. Journal of Nanjing Agricultural University,2013,36(4):111-115(in Chinese with English abstract).
[14] 王林,束建花,赵宁,等. 牛奶和牛血浆中头孢喹肟HPLC-MS/MS检测方法的建立[J]. 畜牧与兽医,2012,44(11):55-59.Wang L,Shu J H,Zhao N,et al. Determination of cefquinome sulfate in milk and bovine plasma by HPLC-MS/MS[J]. Animal Husbandary and Veterinary Medicine,2012,44(11):55-59(in Chinese).
[15] 王延映,彭麟,余祖功. 高效液相色谱发测定饲料中甲硝唑、地美硝唑和异丙硝唑[J]. 南京农业大学学报,2014,37(5):123-127. DOI:10.7685/j.issn.1000-2030.2014.05.020.Wang Y Y,Peng L,Yu Z G. Determination of metronidazole,dimetridazole and ipronidazole in feeds by HPLC[J]. Journal of Nanjing Agricultural University,2014,37(5):123-127(in Chinese with English abstract).
[16] 张伟,彭麟,江善祥. 超高效液相色谱-串联质谱法测定畜禽饲料中氯霉素类药物含量[J]. 南京农业大学学报,2015,38(3):453-458. DOI:10.7685/j.issn.1000-2030.2015.03.013.Zhang W,Peng L,Jiang S X. Determination of choramphenicols in the livestock and poultry feeds by HPLC-MS/MS[J]. Journal of Nanjing Agricultural University,2015,38(3):453-458(in Chinese with English abstract).
[17] 林绍艳,张芳,徐颖洁. 植物中多胺含量超高效液相色谱法的建立[J]. 南京农业大学学报,2016,39(3):358-365. DOI:10.7685/jnau.201511005.Lin S Y,Zhang F,Xu Y J. The establishment of UPLC method for mesuring polyamines content in plants[J]. Journal of Nanjing Agricultural University,2016,39(3):358-365(in Chinese with English abstract).
[18] Raju T S,Kalyanaraman L,Reddy V V,et al. Development and validation of an UPLC method for the rapid separation of positional isomers and potential impurities of rivastigmine hydrogen tartrate in drug substance and drug product[J]. Journal of Liquid Chromatography and Related Technologies,2012,35(7):896-911.
[19] 王志兵,王月,孟庆娟,等. 硅胶固载离子液体-基质固相分散/高效液相色谱法测定禽肉与肝脏中8中磺胺类药物残留[J]. 分析测试学报,2013,32(9):1044-1049.Wang Z B,Wang Y,Meng Q J,et al. Determination of sulfonamide residues in poultry and liver by silica-supported ionic liquid-based matrix solid phase dispersion coupled with high performance liquid chromatography[J]. Journal of Instrumental Analysis,2013,32(9):1044-1049(in Chinese with English abstract).
[20] Shen C L,Chen R N,Qian Z Y,et al. A HPLC-MS/MS method for the quantitation of free,conjugated,and total HDND-7,a novel hesperetin derivative,in rat plasma and tissues:application to the pharmacokinetic and tissue distribution study[J]. Journal of Pharmaceutical and Biomedical Analysis,2016,18(25):149-160.

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备注/Memo

备注/Memo:
收稿日期:2016-12-14。
基金项目:江苏高校优势学科建设工程资助项目(PAPD);校企横向合作项目
作者简介:赵婕,硕士研究生。
通信作者:鲍恩东,教授,博导,主要从事兽医病理学、新兽药创制及药物病理学方面的研究,E-mail:b_endong@njau.edu.cn。
更新日期/Last Update: 2017-07-07