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In chromatography, the retardation factor (R) is the fraction of an analyte in the mobile phase of a chromatographic system. [1] In planar chromatography in particular, the retardation factor R F is defined as the ratio of the distance traveled by the center of a spot to the distance traveled by the solvent front. [2]
The CRFs in thin layer chromatography characterize the equal-spreading of the spots. The ideal case, when the RF of the spots are uniformly distributed in <0,1> range (for example 0.25,0.5 and 0.75 for three solutes) should be characterized as the best situation possible.
The response factor can be expressed on a molar, volume or mass [1] basis. Where the true amount of sample and standard are equal: = where A is the signal (e.g. peak area) and the subscript i indicates the sample and the subscript st indicates the standard. [2]
Paper chromatography is a useful technique because it is relatively quick and requires only small quantities of material. Separations in paper chromatography involve the principle of partition. In paper chromatography, substances are distributed between a stationary phase and a mobile phase.
In contrast to the similar concept called Retention uniformity, R d is sensitive to R f values close to 0 or 1, or close to themselves. If two values are not separated, it is equal to 0. For example, the R f values (0,0.2,0.2,0.3) (two compounds not separated at 0.2 and one at the start ) result in R D equal to 0, but R U equal to 0.3609.
Thin-layer chromatography (TLC) is a chromatography technique that separates components in non-volatile mixtures. [ 1 ] It is performed on a TLC plate made up of a non-reactive solid coated with a thin layer of adsorbent material. [ 2 ]
Planar chromatography is a separation technique in which the stationary phase is present as or on a plane. The plane can be a paper, serving as such or impregnated by a substance as the stationary bed (paper chromatography) or a layer of solid particles spread on a support such as a glass plate (thin-layer chromatography).
The relative mobility (called Rf value or Rm value) is defined as the distance migrated by the protein band divided by the distance migrated by the buffer front. The distances are each measured from the beginning of the separation gel. The migration of the buffer front roughly corresponds to the migration of the dye contained in the sample buffer.