Shodex reversed-phase chromatography tutorial

by | Jul 10, 2018

A general overview of the most used separation mode for HPLC, namely reversed-phase chromatography and a look at the corresponding columns.

In this video one of the most used separation modes -  the reversed phase is being described.

Shodex_RP2The separation is using different polarity of the stationary phase and the mobile phase. The polarity of the stationary phase is lower than that of the mobile phase. Typically the mobile phase contains a mixture of organic solvents (methanol, acetonitrile or THF) and aqueous solvents (water or buffer).

Using the lower polarity mobile phase causes a faster elution. The stationary phase of the HPLC columns can be silica based or polymer based. The advantages of polymer- based columns compared to silica ones are mentioned, like chemical stability full pH-range and many more.
Polymeric columns can use typical functional groups in reversed-phase mode like C-18 group or, and this is different to silica-based columns, can also use no functional group and can create the polarity changed by high or non-polar polymers itself. As polymers polyhydroxymethacrylate, polyvinyl alcohols and styrene divinylbenzene copolymers are often used. The polymer based columns are fully LC/MS compatible.

You can view the video by clicking on the play button in the image below...


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