TLC retardation factor ($R_f$)

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.