High liquid performance chromatography

Question 1

HPLC Pump Requirements

Answer 1

The mobile solvent flows a column, at a rate of .4-1ml/min. The most popular and widely used pump is the resonating pump which is made out of stainless steel to protect form corrosive oxidizing agents. It also sustains pressure up to 5000pi.

Question 2

Injector

Answer 2

Consists of an injection part where the sample is injected once it’s turned. The sample is then pushed into the injector, which moves through the loop, through the column to the detector on the other end. Anywhere between 1-100ul solutes can be added to injector. Samples can be added automatically from multiple being placed on a tray.

Question 3

Analytical column

Answer 3

The column consists of 10cm, 15cm, and 25cm columns with a diameter of 4.6mm-5mm. Analytical columns function more efficiently becuase their shorter diameters allow for less mobile phase solvents to be added, quicker time for particles to be seperated, more particles to be released and as a result increased seperation and resolution times.

Question 4

Precolumn (Guard Column)

Answer 4

Placed above analytical columns and used to protect them form adsorbants that are too strong. Has the same diameter as the analytical column

Question 5

General HPLC Column

Answer 5

Made of stainless steel and ranges anywhere from 5cm to 50cm.

Question 6

Detector

Answer 6

the detector works to translate changes in the amount of analytes passing through the column in electrical signals. There multiple different types of detectors and the type of detector is chosen based on how the specific solute is going to be released, how expensive the detector is, and how powerful it’s ability to detect needs to be. The electrical signals are transmitted through a unit known a a flow cell before reaching the detector (whats a flow cell?). More than one type of HPLC detector can be used to increase the ability to diffrential between multiple different types of analytes comming out of the column and increase the ability to detect more than one type of analyte coming out of the column. The different types of detectors are Refractive index detectors, UV-vis detectors, Flouresecence detectors and electrochemical detectors.