新型苯硼酸材料的制备及其在生物样品中的应用

发布时间：2018-03-22 02:39

本文选题：硼亲和色谱　切入点：核苷　出处：《兰州大学》2017年博士论文　论文类型：学位论文

【摘要】：顺式二羟基类化合物(CDB)是一类重要的生命物质,通常包括糖、糖蛋白、儿茶酚胺、核苷等,它们与众多的生命现象息息相关,在生命科学研究领域中受到广泛关注。但是,在生物样品分析中,它们的含量很低,且基质干扰严重,直接分析面临着挑战。因此,需要发展一种简单有效的富集方法。其中硼亲和材料对于CDB可以实现可逆结合以及专一性识别,从而在顺式二羟基分子的分离富集中发挥着举足轻重的作用。但是,目前硼亲和材料的发展仍然存在一些亟需解决的问题。例如,材料的制备成本高、制备过程复杂、重复利用率低、有效官能团的嫁接量少以及富集容量低等。针对这些问题,我们从以下几个方面进行解决。首先,我们制备了苯硼酸修饰的磁性凹凸棒(ATTA-Fe3O4-NH2-DFFPBA),将磁性纳米粒子和凹凸棒结合,使得磁性材料具有了较大的比表面积,为后续修饰提供了更多的活性位点,并且让凹凸棒带有磁性,便于分离。制备得到的吸附剂对腺苷的富集容量高达13.78 mg g-1,可在高浓度干扰物的混合液中选择性萃取CDB。此外,该硼亲和吸附剂成功用于人尿液中核苷的富集。但是,即使材料具备高比表面积,有限的嫁接量仍然限制了其吸附性能。针对有效官能团嫁接量少的问题,我们采用原子转移自由基聚合(ATRP)技术,在凹凸棒表面聚合链状官能团,再通过引入金纳米粒子增加材料的比表面积,最后通过金和巯基的作用,将巯基苯硼酸嫁接到材料表面。制备得到的ATTApoly(AEMA)-Au-MPBA的富集容量可高至30.83 mg g-1,高于很多文献报道的类似材料。相比苯硼酸修饰的磁性凹凸棒,本工作在富集容量方面有了很大提升。在固相萃取中,粉末材料需要复杂耗时的离心分离过程。于是我们利用相转移沉积技术,结合苯硼酸修饰的SBA-15和磁性聚砜材料,制备了磁性聚砜颗粒(S-IPBA@mPSF)。聚砜材料表面微孔结构减小了传质阻力,有利于分析物从表面向其内部扩散,进而和SBA-15上的有效官能团作用。同时,聚砜高度交联的结构避免了粉末材料的泄露。同时Fe3O4的引入,使材料易于分离,且重复使用率高,可至少重复使用5次。在萃取过程中,加入表面活性剂十六烷基三甲基溴化铵(CTAB),它们通过疏水相互作用聚集在吸附剂表面,为材料提供良好的捕捉介质,从而大大增加了富集倍数,提高了方法的灵敏度。在实际应用中,该方法可以有效萃取出细胞中核苷。该工作虽然实现了材料的重复使用,但是其制备过程复杂、耗时,为了解决这一问题,我们使用商品化的滤纸作为基质,首先在其表面沉积一层非多孔SiO2以增加材料的机械强度,再在其表面进一步沉积一层多孔SiO2来增加材料的比表面积,最后在多孔SiO2表面嫁接上苯硼酸。该吸附剂具有制备成本低、操作简单易行,由于滤纸形状和尺寸的可变性,在其他吸附材料制备方面有很大的应用潜能。上述工作解决了传统吸附材料制备成本高、操作复杂的缺点,但是,要完全依靠人工操作来完成。于是,我们以三聚氰胺海绵为基质,在其表面沉积氧化石墨烯(GO),一方面增加材料的比表面积,另一方面,为后续修饰提供有效官能团,最后通过酰胺化反应嫁接上苯硼酸。在萃取的过程中,把海绵装进固相萃取柱中,然后利用蠕动泵进行分析物的富集和洗脱,实现了半自动化操作。该方法较减少了人工操作,GO的引入也提高了富集容量。以海绵为基质的吸附剂也可以装到预柱中,实现全自动化操作。
[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.

【学位授予单位】：兰州大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：TQ424;O652.6

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