Hepato-neuro lab

HPLC

 Chromatographic technique used to analysis complex samples.

Chromatographic technique used to analysis complex samples, such as cerebrospinal fluid or blood.
First, we treat the sample, by chemical reaction, to add a fluorophore molecule. Then the sample is loaded on a column coated with a hydrophobic compound (molecules could bind to them). Aqueous buffer flows through the system without affecting the protein-column connections. Thereafter, we introduce a hydrophobic buffer gradually. The molecules from the hydrophobic buffer compete molecular interactions in the column, taking out the sample. Fluorescence will be read by a detector. As percentage of second buffer increases with time, interactions will be destroyed gradually, allowing separation of sample molecules (they all have different properties affecting this link).

Typical HPLC chromatogram. Each peak represents a different substance (here, amino acids) while the height of the peak reflects its amount.
Typical HPLC chromatogram. Each peak represents a different substance (here, amino acids) while the height of the peak reflects its amount.

Data obtained by Mélanie Tremblay