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If two bands are not fully resolved in a reverse-phase liquid chromatography experiment when using a...

If two bands are not fully resolved in a reverse-phase liquid chromatography experiment when using a mobile phase of 50:50 water: acetonitrile, which of the two components of the mobile phase should you increase the percentage of in order to improve the resolution? Explain how this change will improve the peak separation?

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Answer #1

Reverse phase chromatography is based on the principle that, stationary phase is nonpolar and mobile phase is Polar.

so as the polar mobile phase moves it takes polar component of the mixture with it and the remaining nonpolar component gets adsorbed on the non polar stationary phase.

in the given condition, if we are not getting a good separation we should increase the polarity of the solvent, that means we should increase the water quantity in the acetonitrile - water mixture.

This will produce effective separation and a better resolution would be obtained.

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