毛细管电泳法应用于头孢菌素类、氟喹诺酮类和磺胺类抗生素药物的检测

密级：学校代码：10075分类号：学号：20090971理学硕士学位论文毛细管电泳法应用于头孢菌素类、氟喹诺酮类和磺胺类抗生素药物的检测学位申请人：左艳丽指导教师：孙汉文教授学位类别：理学硕士学科专业：分析化学授予单位：河北大学答辩日期：二○一二年六月Classified Index: CODE:10075 U.D.C: NO:20090971A Dissertation for the Degree of M.Science.Determination of Some Sulfonamides, Fluoroquinolones and Cephalosporins Drugs by Capillary ElectrophoresisCandidate: Zuo Yan LiSupervisor: Prof. Sun HanwenAcademic Degree Applied for: Master of ScienceSpecialty: Analytical ChemistryUniversity:Hebei UniversityDate of Oral Examination:June, 2012摘要抗生素的大量滥用已经严重威胁到人们的身体健康，专家指出，经常食用含有抗生素的食品，即使是微量的，也能使人出现荨麻疹或造成过敏性休克，而且还会使某些菌株产生耐药性，从而带来预防与治疗某些人畜疾病的困难。

世界各国均对抗生素的使用量提出严格标准，欧盟的标准比日本和美国的标准更加严格，限量值更低，为0.1mg/kg。

我国要想保障本国食品安全并出口产品到上述国家，就必须加强我国自身的检测能力。

本论文不仅应用毛细管区带电泳法对人体液中的药物进行分析，还结合加速溶剂萃取这种新型的样品前处理方法对环境样品和水产品中抗生素残留进行了测定。

第一章：简要介绍毛细管电泳技术、加速溶剂萃取技术及其应用，总结了头孢菌素类、磺胺类、氟喹诺酮类抗生素药物的检测研究。

第二章：建立毛细管区带电泳法同时测定人尿液中的头孢曲松钠和左氧氟沙星含量的新方法。

在优化的条件下，头孢曲松钠和左氧氟沙星具有良好的线性关系（r > 0.999），检出限分别为2.5和7.5 μg·m L-1，加标回收率不小于98.71%，RSD在0.20%到2.00%之间。

该方法准确、快速、简便，可用于临床用药后的尿药浓度检测，也为临床用药和药代动力学研究提供了依据。

第三章：建立了一种加速溶剂萃取(ASE) - 毛细管电泳检测土壤和底泥中磺胺类药物残留的新方法。

在最佳的加速溶剂萃取和毛细管电泳检测条件下，所建立的方法溶剂用量少，回收率高，线性范围宽，方法简便快速,可检测土壤和底泥样品中磺胺间二甲氧基嘧啶、磺胺甲基嘧啶和磺胺嘧啶分别在≥0.070、≥0.063、≥0.056 mg/kg水平上的药物残留。

第四章：建立了一种加速溶剂萃取(ASE)–毛细管电泳法检测水产品中氟喹诺酮类和磺胺类药物残留的新方法。

通过优化萃取条件和分离条件，对样品中七种药物残留进行了测定。

与文献中已有的方法相比，此方法简单，快速，高效，可应用于对水产品中FQs和SAs药物残留的检测。

关键词毛细管电泳；加速溶剂萃取；氟喹诺酮；磺胺类药物；水产品AbstractAbuse of many antibiotics has a serious threat to people's physical health, experts point out that, often eat the food with antibiotics, even trace, also can make a person appear urticaria or cause allergic shock, and still can make certain strains of bacteria resistant to bring forth the difficulties of prevention and treatment of some people and livestock disease. The countries all over the world to use of antibiotics put forward strict standards, the European Union's standard is more strict than the Japanese and Americans, the limited value is 0.1 mg/kg. We have to strengthen our own testing ability in order to ensure our food safety and export products to the above country,This paper not only study drugs in body fluids of infected people by capillary zone electrophoresis method, but also establish the method to determine antibiotic residues in environmental samples and aquatic products with a new type of sample preparation methods- accelerated solvent extraction(ASE).Chapter one: The capillary electrophoresis technique, accelerated solvent extraction and application were described simply. The analytical research and development status of cephalosporin, sulfonamides and fluoroquinolones were reviewed.Chapter two: A novel capillary zone electrophoresis (CZE) method is, for the first time, developed for effective separation and simultaneous determination of ceftriaxone sodium and levofloxacin in human urine. The electrophoresis conditions were investigated and optimized. Good linearity of quantitative analysis was achieved with the correlation coefficients (r) of >0.999. The limits of detection (LOD) for ceftriaxone sodium and levofloxacin were 2.5 and 7.5 μg mL–1, respectively. Their average recovery from human urine was 99.15 % with the RSD of 0.65–1.54 % for ceftriaxone sodium, and 99.57 % with the RSD of 0.2–2.0% for levofloxacin. The proposed method is simple, rapid and accurate, and provides the sensitivity and linearity necessary for selective and simultaneous analysis of the test drugs in human urine at clinically relevant concentrations.Chapter three: A new accelerated solvent extraction (ASE) capillary electrophoretic method was developed for determination of sulfonamide residues in soil and bottom sediment. The method provides a rapid and simple extraction procedure with high recoveries, and has been applied for sample pretreatment and determination of sulfonamide residues in soil and bottom sediment.Chapter four: A rapid and effective accelerated solvent extraction (ASE)–capillary electrophoretic method was developed for the determination of fluoroquinolone and sulfonamide residues in aquatic products. The electrophoresis and extraction conditions were investigated and optimized. The method provides a rapid and simple extraction procedure with high recoveries, and has been applied for the determination of 4 fluoroquinolone and 3 sulfonamide residues in aquatic products with satisfactory results.Key Words Capillary Electrophoresis Accelerated Solvent Extraction Fluoroquinolone Sulfonamide Residues Aquatic Products目录第1章绪论 (1)1.1 毛细管电泳技术概述 (1)1.2 加速溶剂萃取技术 (1)1.2.1加速溶剂萃取技术的原理及优点 (1)1.2.2 加速溶剂萃取技术的应用 (2)1.3头孢菌素类、磺胺类和氟喹诺酮类抗生素的检测研究 (3)第2章毛细管区带电泳法同时测定人尿液中的头孢曲松钠和左氧氟沙星的含量 (5)2.1 引言 (5)2.2 实验部分 (6)2.2.1 试剂与溶液 (6)2.2.2 仪器 (7)2.2.3 尿样处理 (7)2.2.4 毛细管电泳工作条件 (7)2.3 结果与讨论 (7)2.3.1 缓冲溶液的影响 (7)2.3.2 分离电压的影响 (8)2.3.3 温度的影响 (9)2.3.4 选择性 (9)2.3.5 线性方程和检出限 (10)2.3.6 准确度与精密度 (10)2.3.7 回收率实验 (11)2.3.8 应用 (11)2.4 结论 (11)第3章加速溶剂萃取-毛细管电泳法测定土壤和底泥中磺胺类药物残留 (13)3.1 引言 (13)3.2实验部分 (14)3.2.1 仪器与设备 (14)3.2.2 试剂 (14)3.2.3 样品处理 (14)3.2.4 实验方法 (14)3.3 结果与讨论 (15)3.3.1 萃取条件的优化 (15)3.3.2 电泳条件的优化 (16)3.3.3 方法特性 (17)3.3.4 样品分析 (19)3.4 结论 (20)第4章加速溶剂萃取-毛细管电泳法分离测定水产品中氟喹诺酮类和磺胺类药物的残留 (21)4.1 引言 (21)4.2 实验部分 (22)4.2.1 仪器与设备 (22)4.2.2 化学药品和试剂 (22)4.2.3 样品前处理 (23)4.2.4 电泳条件 (23)4.3 结果与讨论 (23)4.3.1 电泳条件的优化 (23)4.3.2 萃取条件的优化 (25)4.3.3 方法特性 (28)4.4结论 (30)参考文献 (31)致谢 (46)攻读硕士期间已发表和待发表论文 (47)第1章绪论1.1 毛细管电泳技术概述毛细管电泳(Capillary Electrophoresis，CE)，又称高效毛细管电泳(High Performance Capillary Electrophoresis，HPCE)，是近二十年迅速发展的一类以毛细管为分离通道、以高压直流电场为驱动力的高效、快速的分离分析技术[1]。