How does hplc works?

Answer 1

HPLC works when a reservoir holds the solvent and then it is sent to the pump manager.Next it goes to the HPLC coloumn .After it goes through there it usually ends in the detector than waste. Generally the stationary phase in the HPLC column is made up of alkyl coated silica making it relatively non-polar. Due to this the technique is also called reversed-phase HPLC.

Answer 2

HPLC (high pressure liquid chromatogaphy) separates unknown compounds based on its polarity. A column is packed with tiny beads, which may have the ability to bind to polar molecules or non polar molecules. Let's say it binds to polar molecules. This is now called a polar stationary phase. We can pour into the column from one end our unknown compounds, and it will slowly pass through the column. Polar molecules travel slower because it is attracted to the polar beads, while the rest will flow right through the other end. Eventually, another solution (mobile phase) can wash off the polar molecules by using a more polar solvent like methanol or acetonitrile. Whatever comes out on the other end can be collected in 1 ml fractions and observed to see how well they are separated.

The reverse phase chromatography has a nonpolar stationary phase.

Answer 3

High-performance liquid chromatography (HPLC) works by separating a mixture of compounds based on their interactions with a stationary phase and a mobile phase. The sample is introduced into the system, where it travels through a column packed with a stationary phase. The compounds within the sample interact differently with the stationary phase, causing them to separate and be detected by a detector.