What effect will the following factors have on a liquid chromatographic separation? (a) column not clamped in a perfectly vertical position; (b) very slow flow rate of the mobile phase; (c) collection of large elution fractions.
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so for a), because the flow is caused by gravity alone, the tilt on the column will cause the sample to unevenly interact with the stationary phase, the largest effect will be on the longitudinal diffusion and bad separation overall.|dw:1443047221529:dw|
b) will allow good equilibration between phases but large band broadening - you'll have a massive, dilute band to collect instead of a tight, concentrated band.|dw:1443047385789:dw|
I'm entirely sure what they mean on c) but i can speculate that the large fractions collected will contain impurities, as well as dilute concentrations of the analyte of intererest - though this shouldn't be a problem if you can evaporate the solvent used but this is not always possible.