【作者】 傅生会；

【导师】 刘绍璞；

【作者基本信息】 西南大学 ， 分析化学， 2008， 硕士

【摘要】 共振瑞利散射（RRS）和共振非线性散射（RNLS）是近年发展起来的新分析技术,因其灵敏度高、仪器价廉、操作简便以及分析快速等优点而引起了人们的广泛关注。在核酸、蛋白质、糖类等生物大分子的测定中已得到了较多的应用,利用该技术在药物分析中的研究工作也日益增多。但未见有应用该技术测定头孢菌素类抗生素的报道。因此,本论文以头孢他啶、头孢曲松钠、头孢哌酮钠和头孢噻肟钠为研究对象,以共振瑞利散射和共振非线性散射光谱法研究了头孢菌素类抗生素与金属离子、染料、表面活性剂之间的相互作用,以及研究了它们的光谱特征、适宜反应条件、影响因素、反应机理和分析应用。1.钯（Ⅱ）-头孢菌素类抗生素螯合物与二溴荧光素相互作用的共振瑞利散射光谱及其分析应用研究在pH为2.8～3.8的Britton-Robinson（BR）缓冲溶液中,头孢他啶（CZD）、头孢曲松钠（CTRX）、头孢哌酮钠（CPZ）和头孢噻肟钠（CFTM）等第三代头孢菌素类抗生素（TGCs）与Pd（Ⅱ）形成1:2的黄色螯合阳离子,它们能进一步与二溴荧光素（DBF）阴离子形成1:3的棕色离子缔合物,不仅引起吸收光谱的变化和荧光猝灭,更导致共振瑞利散射（RRS）的急剧增强并产生了新的RRS光谱。4种抗生素的反应产物有相似的RRS光谱特征,最大RRS波长均位于291nm附近,它们的线形范围和检出限分别为0.0065～1.0μgmL^-1和2.0 ng mL^-1（CZD）;0.0070～1.1μg mL^-1和2.2 ng mL^-1（CTRX）;0.0090～1.6μg mL^-1和2.7 ng mL^-1（CPZ）;0.014～2.2μg mL^-1和4.2 ng mL^-1（CFTM）。研究了适宜的反应条件和共存物质的影响,讨论了离子缔合物的组成,基于离子缔合物反应,发展了一种用RRS技术的高灵敏、简便快速测定TGCs的新方法。2.钯（Ⅱ）-头孢曲松螯合阳离子与阴离子表面活性剂相互作用的共振瑞利散射、二级散射和倍频散射光谱及其分析应用在pH为1.8～2.9的Britton-Robinson（BR）缓冲溶液中,头孢曲松钠（CTRX）与Pd（Ⅱ）形成1∶2的螯合阳离子,它能进一步与十二烷基硫酸钠（SLS）、十二烷基苯磺酸钠（SDBS）和十二烷基磺酸钠（SDS）等阴离子表面活性剂（AS）反应形成1∶3的离子缔合物,导致共振瑞利散射（RRS）、二级散射（SOS）和倍频散射（FDS）的显著增强。最大RRS、SOS和FDS峰分别在335 nm、560 nm和390 nm附近,在一定范围内散射强度与头孢曲松钠的浓度均呈良好的线性关系。用SLS、SDBS和SDS体系测定头孢曲松钠的检出限（3σ）分别为1.8 ng mL^-1、2.3 ng mL^-1和2.3 ng mL^-1（RRS法）,4.9 ng mL^-1、7.4 ng mL^-1和4.7 ng mL^-1（SOS法）以及6.8 ng mL^-1、7.3 ng mL^-1和9.1 ng mL^-1（FDS法）,其中以RRS法灵敏度最高.文中重点研究了RRS法适宜的反应条件和影响因素,对离子缔合物的组成和离子缔合反应机理进行了讨论,考察了共存物质的影响,表明方法有良好的选择性,据此发展了用钯（Ⅱ）和阴离子表面活性剂的以RRS技术灵敏、简便、快速测定头孢曲松钠的新方法.3.Cu（Ⅱ）-头孢曲松-赤藓红离子缔合物的吸收、荧光和共振瑞利散射光谱及其分析应用在pH为4.4～5.1的Britton-Robinson（BR）缓冲溶液中,头孢曲松钠（CTRX）与Cu（Ⅱ）及赤藓红（Ery）形成1∶2∶3的离子缔合物,可引起体系吸收光谱的变化、荧光猝灭和共振瑞利散射（RRS）的急剧增强,并产生了新的RRS光谱,最大RRS波长位于329 nm处。吸光度增加、荧光猝灭和RRS增强在一定CTRX浓度范围内成正比,三者的线性范围和检出限分别为4.4×10^-7～4.0×10^-6g mL^-1和1.3×10^-7g mL^-1（分光光度法）;2.4×10^-7～1.5×10^-6g mL^-1和7.2×10^-8gmL^-1（荧光猝灭法）及2.9×10^-8～2.0×10^-6g mL^-1和8.8×10^-9g mL^-1（RRS法）。确定了适宜的测试条件,讨论了离子缔合物的组成和反应机理。考察了共存物质对RRS法测定CTRX的影响,表明RRS法测CTRX不仅灵敏度高,而且选择性好,用于血清和尿样中头孢曲松钠的测定,取得满意结果。4.HgCl₂-头孢噻肟螯合阴离子与碱性三苯甲烷类染料相互作用的共振瑞利散射光谱及其分析应用在pH为5.3～6.8的Britton-Robinson（BR）缓冲溶液中,头孢噻肟钠（CFTM）与HgCl₂形成1∶1的螯合阴离子,它能进一步与结晶紫、甲基紫、乙基紫、亮绿、碘绿、甲基绿和孔雀石绿等碱性三苯甲烷类染料反应形成三元离子缔合物,导致共振瑞利散射（RRS）的显著增强。最大RRS分别位于367 nm、367 nm、340 nm、367 nm、340nm、340 nm和340 nm附近,在一定范围内散射强度与头孢噻肟钠的浓度均呈良好的线性关系。用结晶紫、甲基紫、乙基紫、亮绿、碘绿、甲基绿和孔雀石绿体系测定头孢噻肟钠的线性范围和检出限（3σ）分别为:0.0089～3.5μg mL^-1和2.7 ng mL^-1,0.0092～3.5μg mL^-1和2.8 ng mL^-1,0.013～3.5μg mL^-1和4.0 ng mL^-1,0.010～3.5μg mL^-1和3.1 ng mL^-1,0.011～3.5μg mL^-1和3.4 ng mL^-1,0.012～4.0μg mL^-1和3.5 ngmL^-1以及0.016～3.5μg mL^-1和4.7 ng mL^-1,其中以结晶紫体系灵敏度最高。文中研究了适宜的反应条件和影响因素,对离子缔合物的组成和离子缔合反应机理进行了讨论,考察了共存物质的影响,表明方法有良好的选择性,据此发展了用HgCl₂和碱性三苯甲烷类染料的灵敏、简便、快速测定痕量头孢噻肟钠的新方法。5.Ce（Ⅳ）与头孢哌酮相互作用的共振瑞利散射、二级散射、倍频散射和荧光光谱及其分析应用在pH为3.5～5.6的HAc-NaAc缓冲溶液中,头孢哌酮（CPZ）与Ce（Ⅳ）发生氧化还原反应,其反应产物再与Ce（Ⅲ）生成的配合物,此时不仅能引起荧光增强,而且能导致共振瑞利散射（RRs）、二级散射（SOS）和倍频散射（FDS）的显著提高。荧光的最大激发(λ_ex)和发射波长(λ_em)分别位于256 nm和349 nm,而最大RRS、SOS和FDS峰分别位于312 nm、550 nm和390 nm附近。在一定范围内荧光强度（△F）和散射强度（△I）与头孢哌酮的浓度均呈良好的线性关系。上述方法测定头孢哌酮的线形范围和检出限分别为0.22-2.5μg mL^-1和6.5 ng mL^-1（FL）;0.0071-2.5μg mL^-1和2.1 ng mL^-1（RRS）;0.018-2.5μg mL^-1和5.4 ng mL^-1（SOS）以及0.022-4.5μg mL^-1和6.6 ng mL^-1。（FDS）,其中以RRS法灵敏度最高。文中重点研究了RRS法适宜的反应条件和影响因素,考察了共存物质的影响,表明方法有良好的选择性,据此通过Ce（Ⅳ）发展了高灵敏、简便、快速测定头孢哌酮的新方法,并用于人血清和尿样中头孢哌酮的定量测定,结果满意。更多还原

【Abstract】 Resonance Rayleigh scattering（RRS）and Resonance nonlinear scattering（RNLS）are new analytical technology developed in recent ten years.For their excellence of high sensitivity,cheap instrument,simple operation and rapid analysis speed,these methods bring to more attention and have been studied widely and applied more and more.At present,these technologies are mainly used to the study of biological macromolecules such as nucleic acids,proteins and saccharide.Resonance Rayleigh scattering and Resonance nonlinear scattering can also be used to determination of some pharmaceuticals.But there are not seen literatures reported to determinate cephalosporin antibiotics by these technologies.Furthermore,in this work,ceftazidime,ceftriaxone,cefoperazone,and cefotaxime are as examples to study the interactions among cephalosporin antibiotics,metal ions and organic ions as such dye,surfactant by RRS and RNLS techniques.Besides,the spectral characteristics,optimum reaction conditions,influencing factors,reaction mechanisms,and their analytical applications have been studied.1.Study on the resonance Rayleigh scattering spectra of the interactions of palladium（Ⅱ）-cephalosporins chelates with 4,5-dibromofluorescein and their analytical applicationsIn pH 2.8-3.8 BR buffer medium,the third generation cephalosporin antibiotics（TGCs）such as ceftazidime（CZD）,ceftriaxone（CTRX）,cefoperazone（CPZ）,and cefotaxime（CFTM）react with palladium（Ⅱ）（Pd（Ⅱ））to form 1:2 yellowish-brown cationic chelates,which further react with 4, 5-dibromofluorescein（DBF）to form 1:3 brown ion-association complexes.As a result,not only the spectra of absorption and fluorescence are changed,but also the resonance Rayleigh scattering（RRS） is enhanced greatly and the new RRS spectra are observed.The four TGCs products have similar spectral characteristics and their maximum RRS wavelengths are all located at 291 nm.The quantitative determination ranges and the detection limits of the four TGCs are 0.0065-1.0μg mL^-1 and 2.0 ng mL^-1for CZD,0.0070-1.1μg mL^-1and 2.2 ng mL^-1for CTRX,0.0090-1.6μg mL^-1and 2.7 ng mL^-1for CPZ,and 0.014-2.2μg mL^-1and 4.2 ng mL^-1for CFTM,respectively.The optimum conditions of the reactions and the effects of foreign substances are investigated,and the composition of ion-association complexes is discussed also.Based on the ion-association reaction,a highly sensitive,simple and rapid method has been proposed to the determination of TGCs.2.Study on the resonance Rayleigh scattering,second-order scattering and frequency doubling scattering spectra of the interactions of palladium（Ⅱ）-ceftriaxone chelate with anionic surfactants and their analytical applicationsIn pH1.8-2.9 Britton-Robinson（BR）buffer medium,ceftriaxone（CTRX）can react with palladium（Ⅱ）（Pd（Ⅱ））to form 1:2 cationic chelate,which can further react with anionic surfactants （AS）such as sodium lauryl sulfonate（SLS）,sodium dodecyl sulfate（SDS）and sodium dodecylbenzene sulfonate（SDBS）to form 1:3 ion-association complexes.As a result,the resonance Rayleigh scattering（RRS）,second-order scattering（SOS）and frequency doubling scattering（FDS） were enhanced greatly.The maximum RRS,SOS and FDS wavelengths of three ion-association complexes were located at 335 nm,560 nmand 390 nm,respectively.The increments of scattering intensity（ΔⅠ）were directly proportional to the concentrations of CTRX in certain ranges.The detection limits（3σ）of CTRX for SLS,SDBS and SDS systems were 1.8 ng ml^-1,2.3 ng ml-1 and 2.3 ng ml^-1（RRS method）,4.9 ng m1^-1,7.4 ng m1^-1and 4.7 ng ml^-1（SOS method）and 6.8 ng m1^-1, 7.3 ng m1^-1and 9.1 ng ml^-1（FDS method）,separately.The sensitivity of RRS method was higher than those of SOS and FDS methods.The optimum conditions of RRS tnethod and the influence factors were investigated,and the composition of ion-association complexes and the reaction mechanism were discussed also.The effects of foreign substances were tested and it showed that the method has a good selectivity.Based on the ion-association reaction,the sensitive,simple and rapid methods for the determination of CTRX have been developed.3.The absorption,fluorescence and resonance Rayleigh scattering spectra of Cu （Ⅱ）-seftriaxone-erythrosin ion-association complex system and their analytical applicationsIn pH 4.4～5.1 Britton-Robinson（BR）buffer solution,ceftriaxone sodium（CTRX）reacts with Cu（Ⅱ）and Erythrosin（Ery）to form 1:2:3 ion-association complex,it results in the change of absorption spectra,the quenching of fluorescence and the great enhancement of Resonance Rayleigh Scattering（RRS）,and the new RRS spectra appear.The maximum RRS wavelength is located at 329 nm.The increment of absorption,the quenching of fluorescence and the enhancement of RRS are all proportional to the CTRX concentration in a certain range.The quantitative determination ranges and the detection limits of the three methods for CTRX are 4.4x10^-7～4.0×10^-6g mL^-1and 1.3×10^-7 g mL^-1（spectrophotometry）,2.4×10^-7～1.5×10^-6g mL^-1and 7.2×10^-8g mL^-1（fluorimetry）,and 2.9×10^-8～2.0×10^-6g mL^-1and 8.8×10^-9g mL^-1（RRS）,respectively.The optimum condition of the experiment was investigated.The composition of ion-association complex and the mechanism of reaction were discussed.Besides,the effects of some co-existing substances determination of CTRX by RRS were investigated.The RRS method has not only high sensitivity but also good selectivity.It can be applied to determination of CTRX in serum and urine samples with satisfactory results.4.Study on the resonance Rayleigh scattering spectra of the interactions of HgCl₂-cefotaxime anionic chelate with basic triphenylmethane dye and their analytical applicationsIn pH 5.3～6.8 Britton-Robinson（BR）buffer medium,Cefotaxime sodium（CFTM）can react with HgCl₂ to form 1:1 anionic chelate,which can further react with basic triphenylmethane dye such as crystal violet,methyl violet,ethyl violet,brilliant green,iodine green,methyl green and malachite green to form ternary ion-association complexes.As a result,the resonance Rayleigh scattering（RRS）was enhanced greatly.The maximum RRS wavelengths of three ion-association complexes were located at 367 nm,367 nm,340 nm,367 nm,340nm,340 nm and 340 nn, respectively.The increments of scattering intensity（△I）were directly proportional to the concentrations of CFTM in certain ranges.The linear ranges and the detection limits（3σ）of CFTM for seven systems were 0.0089～3.5μg mL^-1and 2.7 ng ml^-1（crystal violet）,0.0092～3.5μg mL^-1 and 2.8 ng mL^-1（methyl violet）,0.013～3.5μg mL^-1and 4.0 ng ml^-1（ethyl violet）,0.010～3.5μg mL^-1and 3.1 ng mL^-1（brilliant green）,0.0 11～3.5μg mL^-1and 3.4 ng mL^-1（iodine green）,0.012～4.0μg mL^-1and 3.5 ng mL^-1（methyl green）,and 0.016～3.5μg mL^-1and 4.7 ng mL^-1（malachite green）, separately.The sensitivity of crystal violet system was higher than those of other six systems.The optimum conditions and the influence factors were investigated,and the composition of ion-association complexes and the reaction mechanism were discussed also.The effects of foreign substances were tested and it showed that the method has a good selectivity.Based on HgCl₂ and basic triphenylmethane dye,the sensitive,simple and rapid method for the determination of trace CTRX has been developed.5.Study on the resonance Rayleigh scattering,second-order scattering and frequency doubling scattering spectra of the interaction of Ce（Ⅳ）with cefoperazone and their analytical applications In pH 3.5～5.6 HAc-NaAc buffer medium,cefoperazone（CPZ）can be oxidized by Ce（Ⅳ）, and the oxidation can further react with Ce（Ⅲ）to form a complex.As a result,not only the spectra of fluorescence（FL）is changed,but also the resonance Rayleigh scattering（RRS）,second-order scattering（SOS）and frequency doubling scattering（FDS）are enhanced greatly.The maximum excitation（λex）and emission（λem）wavelengths are at 256 nm and 349 nm,respectively.The maximum RRS,SOS and FDS wavelengths are located at 312 nm,550 nm and 390 nm,respectively. The increments of fluorescence intensity（ΔF）and scattering intensity（Δ1）were directly proportional to the concentrations of CPZ in certain ranges.The linear ranges and the detection limits（3σ）of CPZ for the proposed methods were 0.22-2.5μg mL^-1and 6.5 ng mL^-1（FL）, 0.0071-2.5μg mL^-1and 2.1 ng mL^-1（RRS）,0.018-2.5μg mL^-1and 5.4 ng mL^-1（SOS）,and 0.022-4.5μg mL^-1and 6.6 ng mL^-1（FDS）,separately.The sensitivity of RRS method is higher than those of FL,SOS and FDS methods.The optimum conditions of RRS method and the influence factors are investigated.The effects of foreign substances are tested and it showed that the method has a good selectivity.Based on Ce（Ⅳ）,the sensitive,simple and rapid methods for the determination of CPZ have been developed.It can be applied to determination of CPZ in serum and urine samples with satisfactory results.更多还原