Biotage Isolera Four - Calculate Gradient from TLC Data

411829-L, Operation April 2012 Page 4-10
4
4.4.2 Calculate Gradient from TLC Data
Use the TLC to Linear Gradient editor or the TLC to Step Gradient editor (requires a Spektra license)
to calculate a purification gradient and get cartridge and sample load recommendations.
TLC to Linear Gradient
The software’s TLC to Linear Gradient editor is used to enter the following parameters obtained
from a TLC procedure:
•R
f
values for the compound of interest and the closest eluting compounds on the leading and
trailing sides of the compound of interest.
• Solvents and their percentages.
The software uses these values to calculate a purification gradient and recommend cartridge type
for your sample mass or vice versa.
1. Select the Method tab in the right-hand panel.
2. Press Optimize... and if the Optimize... dialog opens, press Open TLC to Linear Gradient
Editor. The TLC to Linear Gradient editor opens.
3. Select the two solvents to be used by selecting the Solvent A and Solvent B text boxes.
Normally the solvent with the lower solvent strength is selected as Solvent A.
4. Select the Solvent Conditions text box and enter the percentage of Solvent B in the TLC
solution.
Figure 4-6. TLC to Linear Gradient Editor
5. Select the Rf Product text box and enter the R
f
value of the product of interest.
6. Enter the R
f
value of the impurity closest below and above the product in the Rf Impurity 1
and Rf Impurity 2 text boxes. A value for Rf Impurity 1 is required.
7. Select the Cartridge text box and select a cartridge type with a loading capacity exceeding
your crude sample mass.
8. To save the data, press OK. The TLC to Linear Gradient editor closes and the software
calculates the purification gradient based on the TLC data and displays it at the Gradient tab.
9. Set up the rest of the method parameters; see “Specify the Method Parameters at the
Parameters Tab” on page 4-4 and “Specify Collection Parameters at the Collection Tab” on
page 4-8.
10. To save the method and reuse it in the future, see “Save and/or Run the Method” on page 4-9.
11. To run a purification using the (saved or unsaved) method, see “Prepare and Run a Purification”
on page 4-15.