【摘要】：毛細(xì)管電泳(Capillary Electrophoresis,CE),是在毛細(xì)管(分離通道)中,分析物在電場(chǎng)下中依據(jù)各個(gè)組成之間的淌度差異或分配行為完成分離檢測(cè)。由于其具有效率高、成本低、小劑量等特點(diǎn),因此,這些年在分析分離領(lǐng)域越來(lái)越受到科研工作者的重視與青睞,另外,不同檢測(cè)器成功與毛細(xì)管電泳技術(shù)聯(lián)用,給CE檢測(cè)技術(shù)的應(yīng)用范圍帶來(lái)更大的空間,經(jīng)過(guò)科研工作者的不懈努力下,毛細(xì)管電泳相關(guān)技術(shù)已經(jīng)在食品、藥物、環(huán)境、生化等領(lǐng)域蓬勃發(fā)展和廣泛應(yīng)用。然而,由于進(jìn)樣量小和檢測(cè)器等的限制,CE的檢測(cè)靈敏度達(dá)不到生物體系中痕量組分檢測(cè)的需求,為了解決這一問(wèn)題,除了改進(jìn)和完善各類檢測(cè)器外,科研工作者們還致力于研究各種在線富集技術(shù)并與CE聯(lián)用,期待能夠進(jìn)一步提高靈敏度,并且更好地完成對(duì)復(fù)雜樣品的檢測(cè)。本文將毛細(xì)管電泳分別與非接觸式電導(dǎo)檢測(cè)器(C4D)和激光誘導(dǎo)熒光檢測(cè)器(LIF)聯(lián)用,通過(guò)探索并建立不同的在線富集體系,進(jìn)一步研究在線富集技術(shù),并且應(yīng)用于不同的實(shí)際樣品中,主要內(nèi)容如下:第一章為緒論,緒論包括CE的發(fā)展綜述、分離模式和進(jìn)樣方法、檢測(cè)技術(shù)、在線富集技術(shù)及應(yīng)用,并介紹了本論文的研究目的和意義。第二章將場(chǎng)放大進(jìn)樣與毛細(xì)管電泳-非接觸式電導(dǎo)檢測(cè)聯(lián)用(FESI-CE-C4D),實(shí)現(xiàn)了五種人工添加劑(亞硝酸鹽、異抗壞血酸鹽、山梨酸鹽硝酸鹽、硫酸鹽)的在線富集和分離檢測(cè)。在最佳條件下,五種物質(zhì)的檢測(cè)限為0.05-1mgL-1(S/N=3)。為了評(píng)估該模型的應(yīng)用潛力,把該模型運(yùn)用到火腿腸中人工添加劑的檢測(cè)當(dāng)中,樣品處理操作簡(jiǎn)單,且回收率結(jié)果良好(91.9%-106.2%)。第三章將毛細(xì)管電泳(CE)-激光誘導(dǎo)熒光(LIF)聯(lián)用,結(jié)合動(dòng)態(tài)pH界面和瞬時(shí)捕獲兩種在線富集技術(shù),成功地實(shí)現(xiàn)了谷胱甘肽的有效富集和分離檢測(cè)。實(shí)驗(yàn)對(duì)在線富集和分離檢測(cè)的各種參數(shù)進(jìn)行了探究,在動(dòng)態(tài)pH界面下,分析物會(huì)被壓縮成一個(gè)狹窄的區(qū)域,同時(shí)被SDS膠束在樣品區(qū)帶和膠束相邊界捕獲,最后被分離。最優(yōu)條件下,還原型谷胱甘肽,甘氨酸,氧化型谷胱甘肽,半胱氨酸,谷氨酸的檢測(cè)限分別為0.01,0.01,0.5,0.7和O.1nM(S/N=3)。對(duì)比經(jīng)典的CZE模式,本實(shí)驗(yàn)采用瞬時(shí)捕獲結(jié)合動(dòng)態(tài)pH連接在線富集模型的富集倍數(shù)為87-430。此外,該方法已成功地應(yīng)用于細(xì)菌(大腸桿菌、鼠傷寒沙門氏菌和金黃色葡萄球菌)和HaCaT細(xì)胞中還原型谷胱甘肽的定量測(cè)定。第四章將毛細(xì)管電泳(CE)-激光誘導(dǎo)熒光(LIF)技術(shù),聯(lián)合膠束掃集-動(dòng)態(tài)pH界面兩種在線富集技術(shù),對(duì)八種氨基酸進(jìn)行了分離檢測(cè)和在線富集。經(jīng)過(guò)對(duì)分離和富集的各種參數(shù)的探究,最佳條件下,精氨酸,賴氨酸,色氨酸,組氨酸,纈氨酸,酪氨酸,絲氨酸,丙氨酸的檢測(cè)限分別為0.07,0.35,0.15,0.20,0.20,0.15,0.33,0.10nM(S/N=3),富集倍數(shù)達(dá)到44-67之間。另外,將該方法應(yīng)用于人體唾液中檢測(cè)氨基酸,獲得令人滿意的回收率。
[Abstract]:Capillary electrophoresis (Capillary Electrophoresis, CE) is the separation and detection of analyte in an electric field based on the mobility difference or distribution behavior between various components in an electric field. Because of its high efficiency, low cost and small dose, these years are becoming more and more scientific research workers in the field of analysis and separation. In addition, the successful application of different detectors to capillary electrophoresis has brought more space for the application of CE detection technology. With the unremitting efforts of researchers, capillary electrophoresis has been developed and widely used in the fields of food, medicine, environment and biochemistry. The detection sensitivity of CE can not reach the requirement of trace component detection in biological system. In order to solve this problem, in addition to improving and improving all kinds of detectors, researchers are also working on various on-line enrichment techniques and combined with CE in order to solve this problem. Detection of complex samples. In this paper, capillary electrophoresis is combined with non-contact conductance detector (C4D) and laser induced fluorescence detector (LIF) respectively. By exploring and establishing different on-line enrichment systems, the on-line enrichment technology is further studied and used in different solid samples. The main contents are as follows: the first chapter is the introduction and the introduction package. In the second chapter, five kinds of artificial additives (nitrite, isoascorbic acid, sorbic acid) have been realized in the second chapter, including field amplification and capillary electrophoresis noncontact conductivity detection (FESI-CE-C4D). On line enrichment and separation detection of salt nitrate and sulfate. Under the best conditions, the detection limit of five substances is 0.05-1mgL-1 (S/N=3). In order to evaluate the application potential of the model, the model is applied to the detection of artificial additives in the ham sausage, the sample processing operation is simple, and the recovery rate is good (91.9%-106.2%). The third chapter is the hair. CE - laser induced fluorescence (LIF) combined with dynamic pH interface and instantaneous capture of two on-line enrichment techniques have successfully realized the effective enrichment and separation detection of glutathione. The experiments have explored various parameters for on-line enrichment and separation detection. Under the dynamic pH interface, the analyte will be compressed into a narrow area. The domain, at the same time, was captured by the SDS micelles at the sample zone and the micelle boundary, and finally separated. Under optimal conditions, the detection limits of glutathione, glycine, oxidized glutathione, cysteine, and glutamic acid were 0.01,0.01,0.5,0.7 and O.1nM (S/N=3) respectively. Compared with the classical CZE model, the instantaneous capture combined with the dynamic pH connection was used in this experiment. The enrichment factor of the line enrichment model is 87-430.. The method has been successfully applied to the quantitative determination of the reduced glutathione in bacteria (Escherichia coli, Salmonella typhimurium and Staphylococcus aureus) and HaCaT cells. In the fourth chapter, the technique of capillary electrophoresis (CE) laser induced fluorescence (LIF), combined micellar sweep collection dynamic pH interface On line enrichment technology, eight kinds of amino acids were separated and on-line enriched. Through the exploration of the various parameters of separation and enrichment, the detection limits of arginine, lysine, tryptophan, histidine, valine, tyrosine, serine and alanine were 0.07,0.35,0.15,0.20,0.20,0.15,0.33,0.10nM (S/N=3) and enriched times under the optimum conditions. The number reached 44-67. In addition, the method was applied to the detection of amino acids in human saliva, and a satisfactory recovery rate was obtained.
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O658.9

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