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Which technique is more susceptible to column overloading, reversed phase HPLC (C18 stationary phase) or normal phase HPLC?
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Well, he usually shows some questions and answers himself right after the lecture on the topic and this was one of those questions. What I am realizing now is that some of these questions, though written in short and concise form, requires explanation in a long way as a way to re-inforce the lecture, rather than testing to see how much the student got out of the lecture. I say that because I didn't remember his explaining the way you explained below using phrases like "liquid-liquid partition Chromatography" and "liquid-solid adsorption Chromatography" which totally cleared it up for me.In fact, unless there are other qualifications to the question (e.g. did the instructor specify a particular normal-phase column), the answer is *not* normal-phase chromatography. You might want to talk to the prof and ask for more detail.
Thanks for the explanation including how normal-phase separations today are carried out.I'm getting a bit pedantic here, but the distinction between normal-phase and reversed-phase chromatography is fundamentally the relative polarity of the stationary vs. the mobile phases. There is nothing in that definition that makes one inherently lower capacity than another.
A more useful comparison might be between liquid-liquid partition chromatography and liquid-solid adsorption chromatography. Reversed-phase separations on many C18 columns can be explained as partition between the bulk mobile phase and a solvated layer on the surface of the stationary phase. That tends to lead to a high phase ratio (i.e., that nice thick solvated layer represents a lot of stationary phase and therefore high loadability. Normal-phase separations on bare silica or alumina are more easily explained as adsorption of analytes onto the polar surface. Because the stationary phase is a surface instead of a layer of liquid on that surface, there is less of it, and therefore limited loadability.
The catch is that many (if not most) normal-phase separations today are carried out on polar bonded phase columns instead of bare silica and are arguably based on partition into a solvated boundary layer rather than on adsorption.
I hear you:)
Note that all of the above are gross oversimplifications of complex phenomena!
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