本文選題：硼親和色譜　切入點：核苷　出處：《蘭州大學》2017年博士論文　論文類型：學位論文

【摘要】：順式二羥基類化合物(CDB)是一類重要的生命物質,通常包括糖、糖蛋白、兒茶酚胺、核苷等,它們與眾多的生命現(xiàn)象息息相關,在生命科學研究領域中受到廣泛關注。但是,在生物樣品分析中,它們的含量很低,且基質干擾嚴重,直接分析面臨著挑戰(zhàn)。因此,需要發(fā)展一種簡單有效的富集方法。其中硼親和材料對于CDB可以實現(xiàn)可逆結合以及專一性識別,從而在順式二羥基分子的分離富集中發(fā)揮著舉足輕重的作用。但是,目前硼親和材料的發(fā)展仍然存在一些亟需解決的問題。例如,材料的制備成本高、制備過程復雜、重復利用率低、有效官能團的嫁接量少以及富集容量低等。針對這些問題,我們從以下幾個方面進行解決。首先,我們制備了苯硼酸修飾的磁性凹凸棒(ATTA-Fe3O4-NH2-DFFPBA),將磁性納米粒子和凹凸棒結合,使得磁性材料具有了較大的比表面積,為后續(xù)修飾提供了更多的活性位點,并且讓凹凸棒帶有磁性,便于分離。制備得到的吸附劑對腺苷的富集容量高達13.78 mg g-1,可在高濃度干擾物的混合液中選擇性萃取CDB。此外,該硼親和吸附劑成功用于人尿液中核苷的富集。但是,即使材料具備高比表面積,有限的嫁接量仍然限制了其吸附性能。針對有效官能團嫁接量少的問題,我們采用原子轉移自由基聚合(ATRP)技術,在凹凸棒表面聚合鏈狀官能團,再通過引入金納米粒子增加材料的比表面積,最后通過金和巰基的作用,將巰基苯硼酸嫁接到材料表面。制備得到的ATTApoly(AEMA)-Au-MPBA的富集容量可高至30.83 mg g-1,高于很多文獻報道的類似材料。相比苯硼酸修飾的磁性凹凸棒,本工作在富集容量方面有了很大提升。在固相萃取中,粉末材料需要復雜耗時的離心分離過程。于是我們利用相轉移沉積技術,結合苯硼酸修飾的SBA-15和磁性聚砜材料,制備了磁性聚砜顆粒(S-IPBA@mPSF)。聚砜材料表面微孔結構減小了傳質阻力,有利于分析物從表面向其內部擴散,進而和SBA-15上的有效官能團作用。同時,聚砜高度交聯(lián)的結構避免了粉末材料的泄露。同時Fe3O4的引入,使材料易于分離,且重復使用率高,可至少重復使用5次。在萃取過程中,加入表面活性劑十六烷基三甲基溴化銨(CTAB),它們通過疏水相互作用聚集在吸附劑表面,為材料提供良好的捕捉介質,從而大大增加了富集倍數(shù),提高了方法的靈敏度。在實際應用中,該方法可以有效萃取出細胞中核苷。該工作雖然實現(xiàn)了材料的重復使用,但是其制備過程復雜、耗時,為了解決這一問題,我們使用商品化的濾紙作為基質,首先在其表面沉積一層非多孔SiO2以增加材料的機械強度,再在其表面進一步沉積一層多孔SiO2來增加材料的比表面積,最后在多孔SiO2表面嫁接上苯硼酸。該吸附劑具有制備成本低、操作簡單易行,由于濾紙形狀和尺寸的可變性,在其他吸附材料制備方面有很大的應用潛能。上述工作解決了傳統(tǒng)吸附材料制備成本高、操作復雜的缺點,但是,要完全依靠人工操作來完成。于是,我們以三聚氰胺海綿為基質,在其表面沉積氧化石墨烯(GO),一方面增加材料的比表面積,另一方面,為后續(xù)修飾提供有效官能團,最后通過酰胺化反應嫁接上苯硼酸。在萃取的過程中,把海綿裝進固相萃取柱中,然后利用蠕動泵進行分析物的富集和洗脫,實現(xiàn)了半自動化操作。該方法較減少了人工操作,GO的引入也提高了富集容量。以海綿為基質的吸附劑也可以裝到預柱中,實現(xiàn)全自動化操作。
[Abstract]:Two cis hydroxyl compounds (CDB) is a kind of important life material, usually including sugar, glycoprotein, catecholamines, nucleosides, and they are closely related to many life phenomena, has attracted wide attention in the field of life science research. However, in the analysis of biological samples, their content is very low, and the matrix interference direct analysis, facing challenges. Therefore, it needs to develop a simple and effective method. The enrichment of boron affinity material for CDB can achieve reversible binding and specificity recognition, thereby separating rich in cis two hydroxyl molecule concentration plays an important role. However, the current development of boron affinity material still exist some urgent the problem. For example, the preparation of materials of high cost, complicated preparation process, low repeated utilization ratio, grafting amount less effective group and enrichment capacity is low. To solve these problems, we in To solve the following aspects. Firstly, we prepared magnetic bump phenyloboricacid modified bar (ATTA-Fe3O4-NH2-DFFPBA), magnetic nanoparticles and attapulgite with the magnetic material has a larger surface area, provide more active sites for subsequent modification, and let the attapulgite with magnetic separation, convenient adsorbent. The prepared for adenosine enrichment capacity up to 13.78 mg g-1, in the mixed liquor of high concentration of interferent selective extraction of CDB. addition, the boron affinity adsorbent successfully for the enrichment of nucleosides in human urine. However, even if the materials have high specific surface area, the limited amount of grafting is still limited for their adsorption properties. Effective grafting less problem, we use the atom transfer radical polymerization (ATRP) technique in attapulgite surface polymerization of chain functional groups, adding materials by introducing gold nano particles The specific surface area, and finally through gold thiol, 4-mercaptophenylboronic acid will be grafted to the surface of materials. The prepared ATTApoly (AEMA) -Au-MPBA enrichment capacity can be up to 30.83 mg g-1, higher than many similar materials reported in the literature. Compared with the magnetic bump phenyloboricacid modified bar, this work has been greatly enhanced in enrichment capacity. In solid phase extraction, centrifugal powder materials need to be complicated and time-consuming separation process. So we use phase transfer deposition technique, combined with phenylboronic acid modified polysulfone SBA-15 and magnetic materials, magnetic particles were prepared by polysulfone (S-IPBA@ mPSF). Polysulfone material surface microporous structure reduces the mass transfer resistance, is conducive to the analysis of material from the surface spread to the internal, and then SBA-15 the effective functional group. At the same time, the structure of highly crosslinked polysulfone to avoid leakage of powder material. At the same time, the introduction of Fe3O4, the material is easy to separate, and heavy The complex high usage rate, can be reused at least 5 times. In the extraction process, adding surfactant sixteen alkyl three methyl bromide (CTAB), they gathered on the adsorbent surface through hydrophobic interaction, capture medium good for materials, thereby greatly increasing the number of times of enrichment, to improve the sensitivity. In practical application, this method can effectively extract the cells in the work of nucleoside. Although the realization of reusable materials, but the preparation process is complex, time-consuming, in order to solve this problem, we use the commercial paper as substrate, first deposited on the surface of a layer of non porous SiO2 to increase the mechanical the strength of the material, and then the surface to deposit a layer of porous SiO2 to increase the surface area of the material, finally on the surface of porous SiO2 on grafting phenylboronic acid. The adsorbent has low preparation cost, simple operation, the filter paper The shape and size of the variability, have great application potentials in the preparation of other adsorption materials. The work to solve the traditional adsorption material of high preparation cost, operation complexity, but to completely rely on manual operation to complete. Then, we used melamine sponge as a substrate, deposited on the surface of graphene oxide (GO), one hand to increase the surface area of the material, on the other hand, to provide effective functional groups for subsequent modification, finally through amidation grafting. Phenylboronic acid in the extraction process, the sponge into solid phase extraction, enrichment and elution and then analysis by peristaltic pump, semi automatically operation. The method is to reduce the manual operation, the introduction of GO also improved the enrichment capacity. With sponge matrix adsorbent can also be loaded into the pre column, to achieve fully automatic operation.

【學位授予單位】：蘭州大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：TQ424;O652.6

【參考文獻】

相關期刊論文 前1條

1 Jiangnan Zheng;Zian Lin;Lan Zhang;Huanghao Yang;;Polydopamine-mediated immobilization of phenylboronic acid on magnetic microspheres for selective enrichment of glycoproteins and glycopeptides[J];Science China(Chemistry);2015年06期

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本文編號：1646730