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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 />

4-46

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