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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]
In high performance liquid chromatography the CRF is calculated from various parameters of the peaks of solutes (like width, retention time, symmetry etc.) are considered into the calculation. In TLC the CRFs are based on the placement of the spots, measured as RF values.
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]
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 ]
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.
High-performance thin-layer chromatography (HPTLC) serves as an extension of thin-layer chromatography (TLC), offering robustness, simplicity, speed, and efficiency in the quantitative analysis of compounds. [1] This TLC-based analytical technique enhances compound resolution for quantitative analysis.
Chromatographic peak resolution is given by = + where t R is the retention time and w b is the peak width at baseline. The bigger the time-difference and/or the smaller the bandwidths, the better the resolution of the compounds.
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