CombiFlash Torrent® User Manual - Isco
CombiFlash Torrent® User Manual - Isco
CombiFlash Torrent® User Manual - Isco
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<strong>CombiFlash</strong> Torrent® <strong>User</strong> <strong>Manual</strong><br />
CAUTION: Bypassing the solid load cartridge too soon during<br />
the purification run may leave desired compounds on the solid<br />
load cartridge. After removing the cartridge from the fluid<br />
path, these compounds cannot be purified.<br />
If you need to use this feature, perform the following to ensure<br />
that you do not leave desired compounds on the cartridge:<br />
1. Run a TLC plate prior to starting the run. Measure the Rf<br />
(retention factor) of the slowest eluting compound of<br />
interest. (The Rf should be between 0.2 and 0.8 — the same<br />
range used by the GRADIENT OPTIMIZER [4.3.3]). Note the<br />
Percent B solvent used to run the TLC plate. If a compound<br />
displays tailing peaks as streaks on TLC, the Rf used<br />
corresponds to the bottom of the streak.<br />
2.Load the Sample in the Solid Load Cartridge and note the<br />
cartridge size. Refer to list below to determine the cartridge<br />
volume. If the cartridge is partially filled with silica, use<br />
the cartridge volume as if the cartridge were completely<br />
filled with silica.<br />
• 5 gram cartridge = 10 mL volume<br />
• 25 gram cartridge = 45 mL volume<br />
• 65 gram cartridge = 110 mL volume<br />
3.Run the sample and observe the flow rate from the Status<br />
Bar in the upper right corner of the main window. As long<br />
as the system is delivering the programmed flow rate,<br />
there is no need to bypass the cartridge.<br />
4.If the flow rate is slower than that programmed, note the<br />
slowest displayed flow rate. Use the equation below to<br />
determine if the cartridge can be bypassed.<br />
T = Cartridge Volume / (Rf * Flow Rate)<br />
Where T is the time in minutes to wait after the system<br />
reaches the %B solvent used to run the TLC plate; Rf is<br />
the retention factor measured in step 1; Cartridge<br />
Volume is the volume in mL from step 2; and Flow Rate<br />
is the current flow rate in mL/min.<br />
5.Observe the current %B concentration in the status bar.<br />
Once the system reaches the %B percentage used for the<br />
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