#26 Weekly application: chromatography

Chromatography is a fundamental analytical technique used to separate mixtures into their individual components. It is widely used in the chemical and pharmaceutical industries and crucial for various stages of drug development, production, and quality control.

How chromatography works

1.      The sample mixture to be separated is dissolved in a fluid (liquid or gas) called the mobile phase.
2.      This fluid carries the mixture through a system (a column) containing a material called the stationary phase.
3.      Separation: Different components of the mixture move at different speeds due to their varying affinities for the stationary phase, leading to their separation.

Chromatography is essential for purifying active pharmaceutical ingredients (APIs) during the manufacturing process. Nearly all proteins (APIs) absorb UV light at 280nm, the primary reason for this being the presence of aromatic amino acids, e.g. histidine. Installing a Kemtrak DCP007-UV photometer at the column outlet allows the presence of API: s to be detected. This allows the user to know when to start and stop collecting the API.

A common problem is getting a non-sharp peak in the elution due to measurement cells hold up volume. Hold up volumes in a system lower the purity of the collected protein because of dilution effects. Luckily the Kemtrak API analyzer can be used with measurement cells with zero dead/hold up volume, ensuring sharp peaks are detected 💡