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Cyno column mechanism

Posted: Wed Oct 31, 2007 5:30 am
by rick1112
Hi

Could any one tell me what is the underlying mechanism behind retention in “cynoâ€

Posted: Wed Oct 31, 2007 2:50 pm
by Mary Carson
I'm not sure about "cyno" columns, but cyano columns have that -C=N: sticking out, which is a fairly polar moity, so they are considered a normal phase column.

(OK, so underscoring an equals sign doesn't really look like a triple bond....)

Posted: Wed Oct 31, 2007 3:49 pm
by philippem
I assume you mean indeed cyano column.
this column can either be used in normalphase mode or reversed phase mode. Compared to C18 phase the cyano phase has a different selectivity. We used it for compounds with high pi electrons content.(aromatics).

philippe

Posted: Mon Nov 05, 2007 7:39 am
by rick1112
Hi

If it can behave as both normal and reverse phase..? in which way I go ahead with method development.??
Could any one tell me the underlying principle of separation…? Philippem mentioned high pi electrons content..may I know how does this factor contribute in separation mechanism???

Posted: Mon Nov 05, 2007 2:00 pm
by tom jupille
"Normal phase" and "reversed phase" refer to the relative polarities of the column and the solvent. If the column is polar and the solvent is non-polar, it's "normal-phase" chromatography. If the column is non-polar and the solvent is polar, it's "reversed-phase" chromatography.

An archetypical normal-phase column would be something like "bare" silica gel. An archetypical reversed-phase column would be something like octadecyl (C18) groups bonded to a silica support.

Most "cyano" columns consist of something like cyanopropyl groups bonded to a silica support, That cyano group has intermediate polarity, so if you use it with something like hexane/methylene chloride mobile phase, you will be doing normal-phase chromatography. If you use it with water/methanol mobile phse, you will be doing reversed-phase chromatography. (I would not recommend using the same physical column because of problems changing over between immiscible solvents).

phillipem was referring primarily to the use of cyano columns in reversed-phase, where the selectivity is somewhat different from that of C18 columns due to secondary interactions with pi and non-bonding electrons. The fundamental retention mechanism is hydrophobicity, just as in any other reversed-phase separation; the secondary interactions "tweak" retention a bit to move peaks a little further apart (or closer together!).

The mechanism in normal-phase is somewhat hazier (in my mind, at least); the simplest explanation is dipole-dipole interactions with analyte molecules.

Of course, all of the above explanations are over-simplified :wink:

If you can, get a copy of "Practical HPLC Method Development" by Snyder, Glajch, and Kirkland ( http://www.lcresources.com/resources/resbooks.html ). That has much more thorough explanations of what's going on.

The bottom line: you first have to decide what kind of chromatography you want to do (reverse- or normal-phase), then select the column, not the other way around