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For a dilute solute distributed between two liquid regions at local chemical equilibrium, define the chromatographic partition coefficient asHere $c_m,c_s$ are concentrations expressed in the same units, $V_m,V_s$ specify accessible volume in the chromatographic mobile phase and immobilized liquid chromatographic stationary phase, respectively, $n$ is total solute amount of substance, and $f_m$ is the fraction of that amount of substance in the chromatographic mobile phase. Assume a constant chromatographic partition coefficient, rapid exchange, unchanged phases and temperature, and no chemical reaction. A chemical equilibrium population still exchanges molecules; it is not permanently divided into two sets.

The amount of substance balance proves the expression for $f_m$. Increasing either $K$ or $V_s/V_m$ reduces the mobile fraction, illustrating why a concentration ratio alone cannot determine retention. For surface adsorption, specify an adsorption storage law instead of assigning an arbitrary liquid volume to the solid.

An original discrete model makes the separation visible. In each identical cell, first establish local chemical equilibrium, then advance only the mobile fraction by one cell; let the stationary fraction wait. Repeat twelve times, without any additional spreading. With $V_s=V_m$, stipulated chromatographic partition coefficients $K_A=1/3$ and $K_B=3$ give $f_A=3/4$ and $f_B=1/4$. Each cycle contributes mean displacement $f_m$ cells, so the mean positions after twelve cycles are nine cells and three cells. Redistribution itself broadens each band: the cell positions have a binomial distribution with parameters $12,f_m$. These dimensionless chosen values illustrate transport, not measurements of any particular substances.

For thin-layer chromatography, define the TLC retardation factor using distances along the development direction:The origin coordinate is $y_0$; use the centre of a resolved spot and a front marked at the end of development. Both distances begin at the origin, never at the plate bottom. The ratio is dimensionless. An immobile spot has $R_f=0$; a spot at the front has $R_f=1$, where separation from other fast components may be poor. This distance ratio is distinct from the chromatographic partition coefficient and from a column retention time.

For an original stylized plate, choose absolute vertical coordinates $y_0=1\mathrm{cm}$, $y_{\mathrm{front}}=11\mathrm{cm}$, $y_A=4\mathrm{cm}$ and $y_B=8\mathrm{cm}$ above the plate bottom. The solvent distance is therefore $10\mathrm{cm}$, while the spot distances are $3\mathrm{cm}$ and $7\mathrm{cm}$:Using $4/11$ or $8/11$ would wrongly include the unused centimetre below the origin. These chosen positions do not represent measured pigments or the protocol's plate dimensions.

The TLC retardation factor depends on the sample, chromatographic stationary phase, eluent composition, and temperature; plate handling and development conditions must also match when comparing runs. The moving wet front and possible surface adsorption require their own transport assumptions. The equilibrium mobile fraction $f_m$ in the chromatographic partition coefficient model is not unconditionally equal to $R_f$; even that model depends on the phase volume ratio. Coincident TLC retardation factors alone never prove chemical identity.