Translating TLC to Column Chromatography Douglas A. L.

Otte 7/27/2013

1) Run a TLC and adjust the Rf so the two spots you would 3) Choose a column with an appropriate diameter1
like to separate are between 0.2 and 0.4. 1,3 If the separation is relatively easy (Δ Rf ≥ 0.2), the square root of
If the separation is more difficult, you can decrease the Rf to the crude mass in milligrams will give you the appropriate
closer to 0.2. If the separation is easier, you can increase diameter.
the Rf to 0.4. If the separation is more difficult (Δ Rf ≥ 0.1), take the crude mass
In general, ΔCV of ≥2.0 will afford clean separation. Given two in milligrams, divide it by 0.4, and then take the square root of that
spots, ΔCV = (1 / Rf of the spot with the lowest Rf value) – number to give you the appropriate diameter.
(1 / Rf of the spot with the highest Rf value).
For difficult separations (i.e. spots with a Δ Rf ≥ 0.1) this means
the two spots must be in the range of 0.1 and 0.3. 4) Decide how much solvent to use
i.e. if spot #1 Rf = 0.4 and spot #2 Rf = 0.3, ΔCV = (1/0.3)- What is the Rf of the last thing you want to collect on your TLC
(1/0.4) = 0.83 = Bad plate? One divided by this Rf value will tell you how many CV's
i.e. if spot #1 Rf = 0.25 and spot #2 Rf = 0.15, ΔCV = (1/0.15)- you'll need to run the entire column. Next, the amount of silica in
(1/0.25) = 2.7 = Good grams divided by ~0.45 will give you a good idea of what one
For easier separations (i.e spots with a Δ Rf ≥ 0.2) this means Column Volume (CV) is in milliliters. The number of CV's needed
the two spots must be in the range of 0.1 and 4.5. to run the column multiplied by the volume for each CV equals the
i.e. if spot #1 Rf = 0.5 and spot #2 Rf = 0.3, ΔCV = (1/0.3)- total solvent needed to run your column.
(1/0.5) = 1.3 = Okay
i.e. if spot #1 Rf = 0.4 and spot #2 Rf = 0.2, ΔCV = (1/0.2)- 5) Decide how much solvent to collect per fraction
(1/0.4) = 2.5 Good Usually 20 to 30 fractions is plenty.
Take the total volume needed to run the column from step #4 and
2) Decide how much silica to use1,2 divide by the number of fractions you would like to collect.
Assuming you are not simply running your crude material
through a plug of silica, consider the following. For straight forward separations, 20 is adequate, for more difficult
If the Δ Rf of the two spots you are trying to separate is ≥ 0.4, separations, 30 may be more appropriate.
use 15x the crude mass.
If the Δ Rf of the two spots you are trying to separate is ≥ 0.2, General Comments:
use 60x the crude mass. Flow rates for glass columns should be ~ 2 inches / minute
If the Δ Rf of the two spots you are trying to separate is ≥ 0.1,
use 150x the crude mass.

(1) Still, W. C.; Kahn, M.; Mitra, A. J. Org. Chem., 1978, 43(14), 2923-2925. (2) Fair, J. D.; Kormos, C. M. J. Chromatogr. A, 2008, 1211, 49-54.
(3) Stevens, W. C.; Hill, D. C. Mol. Divers. 2009, 13(2), 247-252.
Translating TLC to Column Chromatography Douglas A. L. Otte 7/27/2013

Things to Remember:
1) We use ΔRf’s because they’re quick to derive
& we’re used to thinking in Rf’s but do not
forget that ΔCV’s dictate separation. ∆Rf = 0.1 ∆Rf = 0.2
2) Rf = distance of spot from the origin /
distance of solvent front from the origin Rf = 0.50
3) CV = 1 / Rf Rf = 0.40 Rf = 0.40
∆CV = 1.3
∆CV = 0.83
Rf = 0.25 ∆CV = 2.5
Rf = 0.30 ∆CV = 2.7 Rf = 0.30
Rf = 0.20
Rf = 0.15

(1) Still, W. C.; Kahn, M.; Mitra, A. J. Org. Chem., 1978, 43(14), 2923-2925. (2) Fair, J. D.; Kormos, C. M. J. Chromatogr. A, 2008, 1211, 49-54.
(3) Stevens, W. C.; Hill, D. C. Mol. Divers. 2009, 13(2), 247-252.