CombiFlash Torrent® User Manual - Isco
CombiFlash Torrent® User Manual - Isco
CombiFlash Torrent® User Manual - Isco
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Section 3 Operation<br />
3.2.1 Liquid Sample Injection<br />
If the sample is soluble in the starting mobile phase, it can be<br />
prepared as a solution and injected onto the column when<br />
prompted during the purification run.<br />
3.2.2 Solid Samples<br />
Some compounds are not soluble in solvents that are<br />
compatible with the chromatography. In addition, the<br />
compounds may have very limited solubility in any solvent,<br />
resulting in sample volumes that are impractical for good<br />
chromatography. The answer in this case is solid sample<br />
introduction. Here the reaction mixture or dissolved sample is<br />
mixed with the same media in the chromatography column.<br />
Some samples of this type may be simply dissolved and placed<br />
into a prefilled cartridge. You can then use the cartridge<br />
immediately or dry it before placing it on the system.<br />
Other samples may need more care. Typically you would<br />
create a mixture of 20% sample load to media (w/w). Then dry<br />
the mixture under conditions that will drive off the solvent<br />
without affecting the compounds of interest. These compounds<br />
remain bound to the media. Once dried, pour the<br />
media/sample mixture into an empty cartridge.<br />
To prepare an empty solid sample cartridge:<br />
1. Ensure the empty cartridge has a bottom frit.<br />
CAUTION<br />
Missing frits may cause equipment damage, UV detection<br />
problems, or increased maintenance. Frits prevent solids from<br />
entering the fluid path.<br />
2. Prepare the media and sample:<br />
a. Dissolve your sample in a minimal amount of a suitable,<br />
volatile solvent.<br />
b. Place the media into the solvent. If using silica gel, a<br />
particle size of 40–60 µm (240–400 mesh) is<br />
recommended. The amount of silica required is about<br />
four to five times the mass of your sample.<br />
3-5