配位作用及样品前处理—场放大进样在毛细管电泳在线富集中的应用研究

发布时间：2018-04-21 11:37

本文选题：毛细管电泳 + 在线富集　；参考：《重庆大学》2014年硕士论文

【摘要】：毛细管电泳（capillary electrophoresis，CE）是一种基于电泳和色谱原理的分析技术。虽然该技术在药物分析领域工作中颇受欢迎，但是检测灵敏度低仍然成为影响其在痕量样品分析中的一个重要制约因素，目前已经报道了多种方法用来改善CE的检测限。扫集技术是CE在线富集技术中常用的一种方法，该方法在与质谱等检测器联用方面存在很大的缺陷，限制了CE在线富集的广泛推广。同时，对脂溶性药物的CE在线富集方面研究也相对较少。因此本文在原有富集方法的基础上，对富集体系做出进一步研究，，并提出了新型的配位毛细管电泳-扫集（CICE-sweeping）在线富集技术以及样品前处理-FASS在线富集联用技术，这两种方法对扩大CE适用范围具有重要的意义。本论文的的具体工作内容如下： 1.以氟喹诺酮类抗生素帕珠沙星、依诺沙星和环丙沙星为分析对象，建立了新型CICE-sweeping在线富集技术。在原有的CICE-sweeping在线富集体系的基础上，引入了EDTA区段和碱性高导区段，利用EDTA与金属药物配合物之间的配位置换作用，在毛细管内构建移动络合边界（moving chelation boundary，MCB）。通过对实验条件进行优化和富集体系验证，结果表明建立的新型CE在线富集技术具有良好的富集效果，较传统的CICE-sweeping法富集倍数可提高5-6倍，三种药物的检测限在0.08-0.20μg/mL之间。实验最终将该方法应用到对兔血清中添加的痕量氟喹诺酮类抗生素进行定量检测，得到的回收率在89-94%之间。 2.在上述建立的新型CICE-sweeping在线富集技术的基础上，对四环素类抗生素土霉素、金霉素以及多西环素进行了在线富集和检测。对体系的分离和富集条件进行了优化，最终计算得到的三种药物的富集倍数在146-295之间。这种方法被应用于人血清中痕量土霉素和多西环素的测定中。 3.本节实验以脂溶性维生素A、D3、K1为分析对象，对样品前处理-FASS在线富集技术进行了改进。实验采用十六烷基三甲基溴化铵（CTAB）和十二烷基聚乙二醇醚（Brij-35）混合胶束为样品基质，在电泳前预先将样品溶解在胶束溶液基质中，进样前首先在毛细管中引入高导硼砂缓冲液和水塞，通过电迁移进样使样品随正电荷胶束大量迁移至毛细管中，随后以CTAB溶液为运行电解质的MEKC模式对样品进行分离和检测。在优化的实验条件下，得到的样品峰峰形良好，富集倍数在167-435之间。该方法简便、灵敏、可靠，极大地简化了样品的前处理过程，提高了分析的效率和检测的灵敏度，实现了对脂溶性药物的在线富集。
[Abstract]:Capillary electrophoresis (CEE) is an analytical technique based on the principles of electrophoresis and chromatography. Although this technique is very popular in the field of drug analysis, low detection sensitivity is still an important limiting factor in trace sample analysis. At present, many methods have been reported to improve the detection limit of CE. Scanning technique is a common method in CE on-line enrichment technology. It has great defects in the combination of mass spectrometry and other detectors, which limits the wide application of CE on-line enrichment. At the same time, the study of CE on-line enrichment of liposoluble drugs is relatively rare. Therefore, based on the original enrichment method, the enrichment system was further studied, and a new on-line enrichment technique of coordination capillary electrophoresis (CE) and sweep scanning chromatography (CICE-sweeping) and sample pretreatment (FASS) on-line preconcentration were proposed. These two methods are of great significance to expand the scope of CE application. The specific work of this paper is as follows: 1. A new online enrichment technique of CICE-sweeping was established with fluoroquinolones including pazufloxacin, enoxacin and ciprofloxacin. On the basis of the original CICE-sweeping on-line enrichment system, the EDTA region and the alkaline high conductivity region were introduced. By using the coordination substitution between EDTA and metal complexes, the moving complexation boundary was constructed in capillaries. The experimental conditions were optimized and the enrichment system was verified. The results showed that the new CE on-line enrichment technique had a good enrichment effect, the enrichment multiple of the three drugs was increased 5-6 times than that of the traditional CICE-sweeping method, and the detection limit of the three drugs was between 0.08-0.20 渭 g/mL. The method was applied to the quantitative determination of trace fluoroquinolones in rabbit serum and the recovery was between 89% and 94%. 2. The tetracycline antibiotics oxytetracycline, chlortetracycline and doxycycline were enriched and detected on line based on the new on-line enrichment technique of CICE-sweeping. The separation and enrichment conditions of the system were optimized and the enrichment ratio of the three drugs was calculated to be between 146-295. This method has been applied to the determination of trace oxytetracycline and doxycycline in human serum. 3. In this section, fat-soluble vitamin A ~ (3 +) D _ (3) C _ (3) K _ (1) was used as an analytical object to improve the on-line enrichment technique of sample pretreatment-FASS. The mixed micelles of cetyltrimethylammonium bromide (CTAB) and dodecyl polyethylene glycol ether (Brij-35) were used as the sample matrix, and the samples were dissolved in the micellar solution matrix before electrophoresis. The high conductivity borax buffer solution and water plug were introduced into the capillary before injection. The samples were transported to the capillary with positive charge micelles by electromigration. Then the samples were separated and detected by MEKC mode with CTAB solution as the running electrolyte. Under the optimized experimental conditions, the peak shape of the sample is good and the enrichment ratio is between 167 and 435. The method is simple, sensitive and reliable, greatly simplifies the pretreatment process of samples, improves the analysis efficiency and detection sensitivity, and realizes the on-line enrichment of liposoluble drugs.
【学位授予单位】：重庆大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R917

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