Your question is too general. The answer depends on molecular target, sample matrix, extraction procedure, and expected concentration. There are as many column technolgies as analytical HPLC columns. Thus, check the Waters, Agilent, and Phenomenex websites....

You can have a look on some courses to explain the main processus of a SPE extraction. You may have to search for commercials that you know to provide an example and hw to use it. i remember that I already found some videos in Youtube. Keep searching.

For example "

POLYINTELL Solid Phase Extraction process - YouTube"

SPE is a form of liquid chromatography, so the solvents, stationary phases, modes (i.e. reverse phase, normal phase), etc. are similar to other forms of liquid chormatography such as HPLC. In SPE, you use a column, which you can purchase in any of the afformentioned modes/sorbents. You'd typically attach the column to a vacuum manifold to facilitate the flow of solvent over the stationary phase.

Typically, SPE consists of 4 steps: conditioning, loading sample, rinsing (not always required), and elution. Conditioning involves using solvent to equilibrate the stationary phase to teh solvent you will be loading. For example, one might use methanol followed by diluent. Loading simply involves flowing your sample through the stationary phase. Rinsing may be performed to remove interferences. Finally, elution utilizes a strong solvent to remove the compounds of interest from teh column thus producing a purified sample at an entriched concetration and/or in a solvent compatible with your method.

This is a symplified overview of the technique, and there is not a universal method that works for everything. Thus, you will need to perform method development work to come up with a procedure that works for the analysis you are trying to perform.

check the literature for which ever SPE column you are using. They are not all the same. Start there. THEN do a study to determine what the best conditions are for your molecule. I have seen a huge difference between results reported utilizing the starting method suggested by the manufacturer and the results generated by a modified method. Changing the eluting solvent can have a major impact on sample recovery. Keep in mind that you want the solvent to be compatible with that in your HPLC method. For example if your column is not compatible with acetonitrile it is not a particularly good idea to use it in your sample prep.

Thanks to all professor for your kind suggestions

You should also consider the pH. In some cases it could be useful to eluate your compound of interest at acidic or basic conditions, especially if you have very good imteractions with your SPE column. Then you can perform a couple of washing steps to get rid of your matrix and afterwards eluate your compound in acidic or basic comditions (depending on stability of your substance and wether it is easier to protonate or deprotonate).

Muhammad,

here you can find a good guide on SPE.

https://www.sigmaaldrich.com/Graphics/Supelco/objects/4600/4538.pdf

regards,

G

Thank you