High-performance liquid chromatography of nucleic acid constituents: Chromatographic examination of novel stationary phases
High-performance liquid chromatography of nucleic acid constituents: Chromatographic examination of novel stationary phases
复制标题
核酸成分的高效液相色谱：新型固定相的色谱检测
DOI：
--
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
C. Horváth
中科院分区：
文献类型：
--
作者：
Z. Rassi;C. Horváth
摘要硅胶键合固定相用于核酸组分的分离及其性质的研究，包括同系物低聚核糖腺苷在静电作用层析和烷基苯类在反相色谱中的应用。保留数据的分析证实了表面的层状分子结构，包括一层通过硅氧烷桥固定在二氧化硅表面的丙基链和附着在碳氢官能团上的极性部分。极性顶层包含弱阳离子和/或疏水结合部位，与水洗脱液接触时水化强烈，并阻止大的生物聚合物进入碳氢化合物亚层。然而，在含水有机洗脱液的非极性小分子的反相色谱中，这一层是可接近的，并产生了弱氢碳键合相的典型保留行为。因此，根据样品和流动相的性质，固定相在温和的洗脱条件下表现出用于生物聚合物层析的“软”相的性质，而在反相色谱中通常采用的条件下表现出用于分离小非极性分子的“硬”相的性质。核酸组分在大多数固定相上的保留遵循双重机制，这是由于洗脱液和固定相表面上的结合位置之间的静电和疏水相互作用的结果。具有上述表面形态的硅质固定相适合于具有大范围分子量至3×106道尔顿的核酸组分的分离，只要载体具有合适的孔尺寸。这一点通过对t-RNApha或多腺苷酸消化产生的混合物以及核糖体RNA和不同形式的pBR322 DNA的分离来证明。
SummarySilica-bonded stationary phases were developed for the separation of nucleic acid constituents and their properties investigated with homologous oligoriboadenylic acids in electrostatic interaction chromatography and with alkylbenzenes in reversed-phase chromatography. Analysis of retention data confirmed the stratified molecular structure of the surface which consist of a layer of propyl chains anchoredvia siloxane bridges to the silica surface proper and of polar moieties attached to the hydrocarbonaceous functions. The polar top layer contains weak cationic and/or hydrophobic binding sites, is strongly hydrated in contact with aqueous eluents and bars the access by large biopolymers to the hydrocarbonaceous sublayer. In reversed-phase chromatography of small non polar molecules with hydro-organic eluents, however, this layer is accessible and engenders a retentive behavior typical for weak hydro-carbonaceous bonded phases. As a result the stationary phases, depending on the nature of the sample and the mobile phase, exhibit the properties of "soft" phases for the chromatography of biopolymers under mild elution conditions and those of "hard" phases for the separation of small non-polar molecules under conditions generally employed in reversed-phase chromatography. The retention of nucleic acid constituents on most of the stationary phases investigated subject to a dual mechanism as a result of the interplay of electrostatic and hydrophobic interactions between the eluites and the binding sites on the stationary phase surface. Siliceous stationary phases having surface morphology described above are suitable for the separation of nucleic acid constituents having widely ranging molecular weights up to 3 × 106 Daltons provided the support has appropriate pore dimensions. This is demonstrated by the separation of mixtures arising from digesting t-RNApha or polyadenylic acids as well as those of ribosomal RNA’s and different forms of the plasmid pBR322 DNA.
在涂有交联聚乙烯亚胺的微粒二氧化硅柱上分离合成寡核苷酸。
影响因子：
2.9
作者：
Lawson,TG;Regnier,FE;Weith,HL
通讯作者：
Weith,HL
通过反相高效液相色谱法纯化聚（A）-信使核糖核酸。
DOI：
10.1016/s0021-9673(01)90908-2
发表时间：
1983
期刊：
Journal of chromatography
影响因子：
--
作者：
Simonian,MH;Capp,MW
通讯作者：
Capp,MW