Chromatography Techniques Flashcards

What is the stationary phase in column chromatography?

Answer: The solid matrix packed inside the column

The stationary phase is the solid matrix (e.g., resin or beads) packed inside the column that interacts with sample components to achieve separation.

In size exclusion chromatography (SEC), which molecules elute first?

Answer: Large molecules that are excluded from the pores

Large molecules are excluded from the pores of the resin and travel around them, eluting first with the void volume.

What does the term 'elution' refer to in chromatography?

Answer: Releasing and collecting separated components from the column

Elution is the process of washing separated analytes off the stationary phase so they can be collected as fractions.

Which type of chromatography separates proteins based on their affinity for a specific ligand?

Answer: Affinity chromatography

Affinity chromatography uses a ligand bound to the stationary phase that specifically binds the target molecule, allowing highly selective purification.

What is the mobile phase in high-performance liquid chromatography (HPLC)?

Answer: The liquid solvent pumped through the column

In HPLC, the mobile phase is the liquid solvent mixture that carries the sample through the stationary phase under high pressure.

In ion exchange chromatography, a cation exchange resin carries which charge?

Answer: Negative charge to attract cations

A cation exchange resin has negatively charged functional groups that attract and retain positively charged cations from the sample.

What is the purpose of equilibrating a chromatography column before loading a sample?

Answer: To set the column's pH and ionic conditions to match the sample

Equilibration establishes the correct pH and ionic strength conditions in the column so that the target molecule will bind efficiently when the sample is loaded.