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  本文選題：毛細(xì)管電泳 + 在線富集��； 參考：《重慶大學(xué)》2014年碩士論文

【摘要】：毛細(xì)管電泳（capillary electrophoresis，CE）是一種基于電泳和色譜原理的分析技術(shù)。雖然該技術(shù)在藥物分析領(lǐng)域工作中頗受歡迎，但是檢測靈敏度低仍然成為影響其在痕量樣品分析中的一個重要制約因素，目前已經(jīng)報道了多種方法用來改善CE的檢測限。掃集技術(shù)是CE在線富集技術(shù)中常用的一種方法，該方法在與質(zhì)譜等檢測器聯(lián)用方面存在很大的缺陷，限制了CE在線富集的廣泛推廣。同時，對脂溶性藥物的CE在線富集方面研究也相對較少。因此本文在原有富集方法的基礎(chǔ)上，對富集體系做出進(jìn)一步研究，，并提出了新型的配位毛細(xì)管電泳-掃集（CICE-sweeping）在線富集技術(shù)以及樣品前處理-FASS在線富集聯(lián)用技術(shù)，這兩種方法對擴(kuò)大CE適用范圍具有重要的意義。本論文的的具體工作內(nèi)容如下： 1.以氟喹諾酮類抗生素帕珠沙星、依諾沙星和環(huán)丙沙星為分析對象，建立了新型CICE-sweeping在線富集技術(shù)。在原有的CICE-sweeping在線富集體系的基礎(chǔ)上，引入了EDTA區(qū)段和堿性高導(dǎo)區(qū)段，利用EDTA與金屬藥物配合物之間的配位置換作用，在毛細(xì)管內(nèi)構(gòu)建移動絡(luò)合邊界（moving chelation boundary，MCB）。通過對實驗條件進(jìn)行優(yōu)化和富集體系驗證，結(jié)果表明建立的新型CE在線富集技術(shù)具有良好的富集效果，較傳統(tǒng)的CICE-sweeping法富集倍數(shù)可提高5-6倍，三種藥物的檢測限在0.08-0.20μg/mL之間。實驗最終將該方法應(yīng)用到對兔血清中添加的痕量氟喹諾酮類抗生素進(jìn)行定量檢測，得到的回收率在89-94%之間。 2.在上述建立的新型CICE-sweeping在線富集技術(shù)的基礎(chǔ)上，對四環(huán)素類抗生素土霉素、金霉素以及多西環(huán)素進(jìn)行了在線富集和檢測。對體系的分離和富集條件進(jìn)行了優(yōu)化，最終計算得到的三種藥物的富集倍數(shù)在146-295之間。這種方法被應(yīng)用于人血清中痕量土霉素和多西環(huán)素的測定中。 3.本節(jié)實驗以脂溶性維生素A、D3、K1為分析對象，對樣品前處理-FASS在線富集技術(shù)進(jìn)行了改進(jìn)。實驗采用十六烷基三甲基溴化銨（CTAB）和十二烷基聚乙二醇醚（Brij-35）混合膠束為樣品基質(zhì)，在電泳前預(yù)先將樣品溶解在膠束溶液基質(zhì)中，進(jìn)樣前首先在毛細(xì)管中引入高導(dǎo)硼砂緩沖液和水塞，通過電遷移進(jìn)樣使樣品隨正電荷膠束大量遷移至毛細(xì)管中，隨后以CTAB溶液為運(yùn)行電解質(zhì)的MEKC模式對樣品進(jìn)行分離和檢測。在優(yōu)化的實驗條件下，得到的樣品峰峰形良好，富集倍數(shù)在167-435之間。該方法簡便、靈敏、可靠，極大地簡化了樣品的前處理過程，提高了分析的效率和檢測的靈敏度，實現(xiàn)了對脂溶性藥物的在線富集。
[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.
【學(xué)位授予單位】：重慶大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R917

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本文編號：1782315