Week 8: High Performance Liquid Chromatograph

Name: Eriko Wong (0700477C)

Section: Analytical Chemistry

Ok, so today I'm going to share with you about the components of HPLC and how does it work.

Here is a picture showing the different components of HPLC::)

Image source: http://www.protein.iastate.edu/hplc.html

1) Mobile Phase Supply System are to be filtered and degassed first before pass through the column so as to remove any small particles or dissolved gases. If this step is not done, it can cause the clogging of column.

2) Injector is where you would inject your sample in. This is done by a syringe and a needle. Sample is also filtered first before being carried to the column for analysis.

3) Packed column: is also known as the stationary phase. This is where the interactions between the sample, mobile phase and stationary phase occur. The different interaction of the different components in the sample will lead to the different time they elute out of the column. Often, a guard column is installed to trap unwanted materials. Column is usually housed in a oven, under a predetermined temperature to ensure a consistent interaction of the sample, mobile and stationary phase.

4) Detector: The one I'm using is Ultraviolet-visible spectrometry detector. At different wavelength, the peak area and height of the sample recorded will be different too.

An example of a chromatogram:: (x-axis: time, y-axis: the absorption by the detection)

Image Source: http://www.protein-ms.de/METHODS/HPLC/HPLC.htm

5) Recorder: This is to obtain a real time plot of the analysis. Once the analysis is done, the computer will print out the data, along with the chromatogram.

Hopefully you guys get the idea of how HPLC works.... :)