柱层析法及其组分的分离方法

描述

色谱法是一种实验室技术用于化学分析分离成其成分混合物。混合溶剂溶解在液体(气体或液体)被称为流动相,它传输通过一个系统(一列、毛细管板或板),是固定的材料称为固定相。因为这种混合物的成份有不同的亲和力为固定相,并保留时间长短不一的根据与表面的交互网站,他们在不同的移动液体表观速度使它们分开。分离是通过微分分区移动和固定的阶段。

分配系数的概念验证色谱法。任何溶质分区两个非混相溶剂之间发生。当我们做一个溶剂不动,另一个手机,我们得到了最常用的色谱应用程序。如果矩阵支持或固定相极性(如纸、硅等)色谱法是前进阶段如果非极性(C-18)反相色谱法。

柱层析法是一种分离方法的固定床是包含在一个管子。固体固定相的粒子或者支持涂上液体固定相填充的整个体积内管(填充柱)或集中或沿管壁内,离开一个开放的、不受限制的路径在中间管的流动相(开放的管状柱)。不同样品的保留时间,计算基于利率的差异通过介质的运动。传统方法非常类似于柱层析法除溶剂是被迫通过列应用正压。现在大多数分离可以完成在不到20分钟,比以前有更好的分离方法。现代闪光色谱系统的形式销售预包装塑料墨盒溶剂注入。系统还可以与探测器和一部分收藏家允许自动化。梯度泵加速分离,减少溶剂消耗。流化床,而不是固相形成的填充层,用于膨胀床吸附。文化的培养基配方或碎料浆细胞这允许省略初始结算步骤如离心和过滤。 A column is made by packing a solid absorbent into a cylindrical glass or plastic tube. The size is determined by the amount of compound isolated. To keep the solid phase in place, the tube's base contains a filter either a cotton or glass wool plug or glass frit. A solvent reservoir can be attached to the top of the column. The dry method and the wet method are the two most common methods for preparing a column. For the dry method, the column is first filled with dry stationary phase powder, followed by the addition of mobile phase, which is flushed through the column until it is completely wet and is never allowed to run dry from this point on. For the wet method, slurry of the eluent and the stationary phase powder is prepared and carefully poured into the column. The top of the silica should be flat and it can be protected with a layer of sand. To advance the organic material, eluent is slowly passed through the column. The stationary phase retains the individual components differently and separates them while they run at different speeds through the column with the eluent. They elute one at a time at the column's end. The eluent is collected in a series of fractions throughout the chromatography process. Fraction collectors are devices that collect fractions automatically. By running multiple columns at once, chromatography productivity can be increased. Multi-stream collectors are used in this case. The eluent flow's composition can be monitored, and each fraction is analyzed for dissolved compounds using analytical chromatography, UV absorption spectra or fluorescence, for example.

柱层析法是利用固体固定相或吸附剂。硅胶是最常用的固定相在氧化铝柱层析法第二。此前,纤维素粉被广泛使用。进行离子交换色谱法、反相色谱(RP)、亲和色谱法或膨胀床吸附(EBA),各种各样的静止阶段是可用的。静止阶段通常是细碎的粉末或凝胶和/或微孔表面积增加,尽管膨胀床吸附(EBA)采用流化床。流动相,也称为洗脱液溶剂或混合溶剂,用于移动通过列化合物。它选择感兴趣的化合物的保留因子值大约是在0.2和0.3之间,以减少所需的时间和数量的洗脱液色谱法。洗脱液也被选中,这样各种化合物可以有效地分离。