【作者】 段慧；

【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）和二级散射、倍频散射等共振非线性散射（RNLS）是二十世纪90年代发展起来的一种新兴的分析技术，因其高灵敏度和简易性而受到人们的广泛关注。目前，RRS和RNLS已经用于生物大分子、无机离子和纳米微粒的研究和测定。该技术在药物分析中的应用也日益增多，但是对青霉素类药物的分析应用研究较少。因此本文以青霉素类抗生素为主要对象，研究和发展了用RRS和RNLS法测定这类药物的新体系和新方法。本文重点研究青霉素类抗生素与金属、某些氧化剂以及染料之间的相互作用的RRS和RNLS光谱特征、适宜的反应条件和影响因素，讨论了反应机理，并建立相应药物的分析方法。主要研究内容如下：1铁氰化钾高灵敏共振瑞利散射法测定青霉素类抗生素在稀HCl介质中，K₃[Fe（CN）₆]与阿莫西林（AMO）、氨苄西林（AMP）、氯唑西林钠（CLO）、羧苄西林钠（CAR）和青霉素钠（BEN）等抗生素药物在加热条件下反应生成结合产物，会导致溶液的共振瑞利散射（RRS）强度急剧增强，并产生新的RRS光谱，5种反应产物的最大散射峰均位于330附近。在一定的浓度范围内，不同的反应体系RRS强度（△I）与药物浓度成正比，反应具有很高的灵敏度，K₃[Fe（CN）₆]对5种药物的检出限（3σ／s）分别在4．61 ng·ml^-1至5．62 ng·ml^-1之间。本文研究了适当的反应条件和影响因素，并考察了共存物质的影响，表明方法具有较好的选择性，可用于胶囊和血清及尿液中青霉素类药物的测定。2钯（Ⅱ）与阿莫西林和氨苄西林相互作用的共振瑞利散射光谱及其分析应用在酸性介质中加热，使阿莫西林（AMO）和氨苄西林（AMP）等侧链含苄氨基的青霉素类抗生素发生降解，其降解产物青霉胺和苄氨基青霉醛在pH5左右的弱酸性介质中能进一步与钯（Ⅱ）反应形成组成比1：1：1的混配型三元配合物，此时将引起共振瑞利散射（RRS）的显著增强，并出现新的RRS光谱。钯（Ⅱ）与两种药物的反应产物具有相似的RRS光谱特征，最大散射波长均位于370 nm。在一定范围内散射增强（△I）与药物的浓度成正比。方法具有较高的灵敏度，对于AMO和AMP的检出限（3σ）分别为18．0和15．4 ng·mL^-1。此时侧链不含苄氨基的其他青霉素不产生类似反应，并且也允许一定量的其他物质存在，因此方法有较好的选择性，可用于胶囊、片剂及血清、尿样中阿莫西林和氨苄西林的测定，能获得较满意的结果。3 Fe₃[Fe（CN）₆]₂纳米微粒共振瑞利散射、二级散射和倍频散射法间接测定青霉素类抗生素在盐酸溶液和加热条件下，阿莫西林（AMO）、氨苄西林（AMP）、氯唑西林钠（CLO）、羧苄西林钠（CAR）和青霉素钠（BEN）等青霉素类抗生素能还原Fe（Ⅲ）而生成Fe（Ⅱ），Fe（Ⅱ）与Fe（CN）₆^3-反应生成深蓝色的Fe₃[Fe（CN）₆]₂，疏水性的配合物Fe₃[Fe（CN）₆]₂能进一步聚集形成平均粒径为45 nm的纳米微粒{Fe₃[Fe（CN）₆]₂}_n，此时不仅会引起吸收光谱的变化，在738 nm附近出现新的吸收峰，对不同的药物其摩尔吸光系数（ε）在（1．0-2．3）×10⁴ L·mol^-1·cm^-1之间，而且更能观察到共振瑞利散射（PRS）、二级散射（SOS）和倍频散射（FDS）等非线性散射的显著增强。5种青霉素的反应产物具有相似的RRS，SOS和FDS光谱特征，其最大RRS，SOS和FDS波长分别位于341，640和390 nm处，但不同药物体系的相对散射强度有一定差异。散射强度（△I）在一定范围内与药物的浓度成正比，方法对于这类青霉素抗生素的检出限分别为2．9-6．1 ng·mL^-1（RRS法）、4．0-6．8 ng·mL^-1（SOS法）和7．4-16．2 ng·mL^-1（FDS法），其中以RRS法和阿莫西林的灵敏度最高。本文主要研究了RRS的反应条件和影响因素，并以阿莫西林为例，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究结果，建立了一种灵敏、简便、快速间接测定青霉素类抗生素的新方法，并用于人血清、尿样和片剂及胶囊制剂中青霉素类抗生素的定量测定。4铈（Ⅳ）与阿莫西林相互作用的共振Rayleigh散射、二级散射和倍频散射光谱及其分析应用在pH5．0的HAc-NaAc缓冲溶液中，铈（Ⅳ）能与阿莫西林（AMO）相互作用，导致共振瑞利散射（RRS），二级散射（SOS）和倍频散射（FDS）光谱显著增强，并出现新的RRS，SOS和FDS光谱，最大散射波长分别位于370，558和390nm，散射强度在一定范围内与AMO的浓度成正比，此方法具有较高的灵敏度，对于AMO的检出限（3σ）分别为2．2ng·mL^-1（RRS）、3．0 ng·mL^-1（SOS）和5．2 ng·mL^-1（FDS）。本文研究了适宜的反应条件和影响因素，并考察了共存物质的影响，表明方法具有较好的选择性，据此发展了一种用铈（Ⅳ）测定AMO的共振散射新方法，并用于胶囊片剂及人血清、尿液中AMO的测定。5 KIO₃-溴化钾-碱性三苯甲烷类染料体系共振瑞利散射法测定青霉素类抗生素在弱酸介质中，阿莫西林（AMO）、氨苄西林（AMP）、羧苄西林钠（CAR）、氯唑西林钠（CLO）和青霉素钠（BEN）等青霉素类抗生素（PENs）的碱水解产物与碘酸钾反应生成I₂，I₂可进一步与溴化物生成[I₂Br]^-配阴离子，它再与甲基紫（MV）、乙基紫（EV）和结晶紫（CV）等碱性三苯甲烷类染料（BTMD）形成离子缔合物[I₂Br][BTMD]，使共振瑞利散射（RRS）显著增强并产生新的RRS光谱，其最大散射波长均位于330nm附近。在一定条件下，散射强度（△I）与青霉素类抗生素浓度成正比，其线性范围分别在0．008-2．0（MV-PENs体系），0．004-1．0（EV-PENs体系）和0．008-2．0μg·mL^-1（CV-PENs体系）之间．方法具有高灵敏度，对五种抗生素药物的检出限（3σ）分别为2．3-4．4（MV-PENs体系），1．3-1．7（MV-PENs体系）和2．4-3．8 ng·mL^-1（MV-PENs体系）。本文研究了适当的反应条件和影响因素，并考察了共存物质的影响，表明方法具有较好的选择性。此法可用于胶囊和片剂等药物制剂以及血清和尿液中青霉素类药物的测定。更多还原

【Abstract】 Resonance Rayleigh scattering （RRS） and resonance non-linear scattering （RNLS） such as second order scattering and frequency doubling scattering etc. is a new technology developed in 1990s. Due to its remarkable characteristics of high sensitivity and simple operation, this method has received much attention. It has been extensively and successfully unitized to study and determination of biological macromolecules, trace metal ions, nonmetal ions and nanoparticle probe et al. Furthermore, it has been applied in pharmaceutical field more and more. However, it is seldom applied to the study of penicillin antibiotics. Therefore, taking the penicillin antibiotics as examples, we have studied and developed the application of the RRS and RNLS methods. In this thesis, we mainly investigated the interaction between penicillin antibiotics and metal ions, oxidants and dyes by the RRS and RNLS methods. Besides, the spectral characteristics, the optimum conditions and the influencing factors were studied. The reaction mechanism was discussed too. Based on the above researches, the RRS and RNLS methods have been established for the determination of penicillin antibiotics. Main investigated systems are listed as follows:1. A Highly Sensitive Resonance Rayleigh Scattering Method for the Determination of Penicillin Antibiotics with Potassium FerricyanideHeated in a boiling water bath, penicillin antibiotics such as amoxicillin, ampicillin, sodium cloxacillin, sodium carbenicillin and sodium benzylpenicillin could react with K₃[Fe（CN）₆] to form combined products in a dilute HCl medium. As a result, resonance Rayleigh scattering （RRS） intensity was enhanced greatly and new RRS spectra appeared. The maximum scattering wavelengths of the five combined products are all located at 330 nm. The scattered intensity increments （△I） of the combined products are directly proportional to the concentrations of the antibiotics in a certain range. The methods exhibit high sensitivity, and the detection limits for the five penicillin antibiotics are between 4.61 and 5.62 ng·mL^-1. The spectral characteristics of RRS and the optimum reaction conditions were investigated. The mechanism of reaction and the reasons for the enhancement of resonance light scattering were discussed. The effects of coexisting substances have been examined, and the results indicated that the method had a good selectivity. It can be applied to the determination of penicillin antibiotics in capsule, tablet, human serum and urine samples.2. Resonance Rayleigh Scattering Spectra of Interaction of Amoxicillin or Ampicillin with Palladium（Ⅱ） and Their Analytical ApplicationsHeated in acidic medium, penicillin antibiotics containing an amino group side chain such as amoxicillin （AMO） and ampicillin （AMP） can be degraded, and the penicillamine and benzyl aminopeniloaldehyde of the degradation products can further react with palladium（Ⅱ） to form a composition ratio 1:1:1 blended ternary complex, which leads to the great enhancement of resonance Rayleigh scattering （RRS） and appearance of new RRS spectra. The spectral characteristics of the reaction products of the two drugs with palladium（Ⅱ） are similar, and the maximum wavelengths are all at 370 run. The scattering intensity increments （△I） of the products are directly proportional to the concentrations of the antibiotics in certain ranges. The method exhibits higher sensitivity, and the detection limits （3σ） for AMO and AMP are 18.0 and 15.4 ng·mL^-1, respectively. In this case, the other penicillin antibiotics without any amino group in the structure of the side chain have not similar reactions, and certain amounts of some coexisting substances have no interference. Hence, the method has a good selectivity. It can be applied to the determination of amoxicillin and ampicillin in capsule, tablet, human serum and urine samples with satisfactory results.3. Resonance Rayleigh Scattering, Second Order Scattering and Frequency Doubling Scattering Methods for the Indirect Determination of Penicillin Antibiotics Based on the Formation of Fe₃[Fe（CN）₆]₃ NanoparticlesUnder the condition of HC1 solution and heating, penicillin antibiotics such as amoxicillin （AMO）, ampicillin （AMP）, sodium cloxacillin （CLO）, sodium carbenicillin （CAR） and sodium benzylpenicillin （BEN） can react with Fe（Ⅲ） to produce Fe（Ⅱ） which further reacts with K₃Fe（CN）₆ to form navy-blue Fe₃[Fe（CN）₆]₂.The hydrophobic Fe₃[Fe（CN）₆]₂ complex will associate to form {Fe₃[Fe（CN）₆]₂}_n nanoparticles with an average diameter of 45 nm, which not only result in the change of absorption spectra and appearance of a new absorption peak at 738 nm with the molar absorptivity （ε） of （1.0-2.3）×10⁴ L·mol^-1·cm^-1 for different antibiotics, but also lead to the significant enhancement of resonance Rayleigh scattering （RRS） and non-linear scattering such as second order scattering （SOS） and frequency doubling scattering （FDS）. The RRS, SOS and FDS spectral characteristics of reaction products for five antibiotics were similar, and the maximum scattering wavelengths were 341nm for RRS, 640 nm for SOS and 390 nm for FDS, respectively. But the relative scattering intensities for the investigated drug systems were different. The scattered intensity increments （△I） of five products were directly proportional to the concentrations of antibiotics in certain ranges. The detection limits for five penicillin antibiotics were 2.9-6.1 ng·mL^-1 for RRS method, 4.0-6.8 ng·mL^-1 for SOS method and 7.4-16.2 ng·mL^-1 for FDS method, respectively. Among them, the RRS method exhibited the highest sensitivity. Moreover, the AMO system was more sensitive than other antibiotics systems by three scattering methods. In this work, the reaction conditions of RRS method and the influencing factors were investigated. And taking AMO as an example, the effects of coexisted substances were examined indicating that the method had a good selectivity. Based on the above researches, a highly sensitive, simple and fast method for the indirect determination of penicillin antibiotics has been developed. It was applied in the determination of penicillin antibiotics in capsule, tablet, human serum and urine samples.4. Resonance Rayleigh Scattering, Second Order Scattering and Frequency Doubling Scattering Spectra of Interaction of Amoxicillin with Cerium （Ⅳ） and its Analytical ApplicationIn pH5.0 acetic acid-sodium acetate buffer solution, the interaction of amoxicillin （AMO） with Ce（Ⅳ） can result in the great enhancement of resonance Rayleigh scattering （RRS） and resonance non-linear scattering （RNLS） such as second order scattering （SOS） and frequency doubling scattering （FDS）. The new spectra of RRS, SOS and FDS appear and their maximum wavelengths are located at 370, 558, 390 nm, respectively. The scattered intensity increments are directly proportional to the concentration of AMO in a certain range. The methods exhibit high sensitivity, and the detection limits （3o） for the determination of AMO are 2.2 ng·mL^-1（RRS method）, 3.0 ng·mL^-1 （SOS method） and 5.2 ng·mL^-1 （FDS method）. In this work, the optimum conditions and the influencing factors were investigated. Meanwhile, the effects of coexisting substances have been examined, and the results indicated that the method had a good selectivity. It can be applied to the determination of AMO with Ce（Ⅳ） in capsule, tablet, human serum and urine samples.5. Determination of Penicillin Antibiotics by Resonance Rayleigh Scattering Method with KIO₃-KBr- Basic Triphenylmethane DyesIn a weak acid medium, the basic hydrolysis product of penicillin antibiotics （PENs） such as amoxicillin （AMO）, ampicillin （AMP）, carbenicillin （CAR）, sodium cloxacillin （CLO） and sodium benzylpenicillin （BEN） reacts with potassium iodate to form I₂ which further interacts with Br^- resulting in the formation of [I₂Br]^- anionic complex. Then an ion-association complex [I₂Br][ BTMD] was formed owing to the interaction of [I₂Br]^- with basic triphenylmethane dyes （BTMD） such as methyl violet （MV）, ethyl violet （EV） and crystal violet （CV）, which led to the significant enhancement of resonance Rayleigh scattering （RRS） and the appearance of new RRS spectra with their maximum wavelengths near at 330 nm. In certain conditions, the intensities of RRS （△I） are directly proportional to the concentrations of penicillin antibiotics in the range of 0.008-2.0 （MV-PENs system）, 0.004-1.0 （EV-PENs system） and 0.008-2.0μg·mL^-1 （CV-PENs system）. The method exhibits high sensitivity, the detection limits are 2.3-4.4 （MV-PENs system）, 1.3-1.7 （MV-PENs system） and 2.4-3.8 ng·mL^-1 （MV-PENs system）, respectively. In this work, the reaction conditions and the influencing factors have been investigated, and the effects of coexisting substances also have been examined indicating a good selectivity of the method. It can be applied to the determination of penicillin antibiotics in capsule and tablet pharmaceutical formulations and human serum and urine samples.更多还原