High-performance liquid chromatography ( HPLC ; formerly referred to as high-pre

Question

High-performance liquid chromatography (HPLC; formerly referred to as high-pressure liquid chromatography), is a technique in analytical chemistry used to separate, identify, and quantify of each component in a mixture.

Reversed-phase chromatography employs a polar (aqueous) mobile phase and the non-polar stationary phase. As a result, hydrophobic molecules in the polar mobile phase tend to adsorb to the hydrophobic stationary phase, and hydrophilic molecules in the mobile phase will pass through the column and are eluted first. The most common nonpolar stationary phases use an organochlorosilane where the R group is an n-octyl (C8) or n-octyldecyl (C18) hydrocarbon chain. In reversed-phase HPLC, the order of elution is the opposite of that in a normal-phase separation, with more polar solutes eluting first. Increasing the polarity of the mobile phase leads to longer retention times. Shorter retention times require a mobile phase of lower polarity. Accordingly, the order of elution in the increasing order of polarity is:

C < B < A

_____________ Increasing polarity

Explanation / Answer

4(6 pointy) Answer the following questions regarding the HPLC expe a) (5 points) The chromatogram be mixture. The system used a reversed phase column and a water-acetonitrile eluent. Ct order of elution,lit the components in order of increasing polarity and clearly explan n the xperiment. low was obtained in the analysis of a multicom ponen choice. minutes

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