Question

What precautions must be taken when you introduce a mixture of compounds to be separated onto a liquid chromatography column?

Answer 1

Precautions must be considered when introducing a mixture of compounds onto a liquid chromatography column to ensure accurate separation.

Step-by-step explanation:

When introducing a mixture of compounds onto a liquid chromatography column, there are several precautions that must be taken to ensure accurate separation. First, the sample should be introduced as a narrow band at the top of the column. This helps to maintain the initial concentration profile and prevent broadening of the bands. Second, the solvents used in the mobile phase should be compatible with the stationary phase and solutes to avoid any interactions that could affect separation. Finally, the column should be equilibrated with the mobile phase before introducing the sample to ensure consistent separation.

Answer 2

The slim exit of the column is first persisted with glass wool or a permeable plate in order to sustain the column packing element and keep it from getting out of the tube. Then the adsorbent solid, which is usually a silica, is firmly packed into the glass tube to make the separating column. The packing of the non-moving phase into the glass column must be done with precaution to create an even distribution of material. An even distribution of adsorbent material is very important to lessen the existence of air bubbles and/or channels inside the column. To finish preparing the column, the solvent to be used as the mobile phase is delivered through the dry column. Then the column is said to be "wetted" and the column must stay wet throughout the entire procedure. Once the column is properly prepared, the sample to be separated is placed at the top of the wet column.