Hello, you purified rather than you extracted fatty acid with prep HPLC. There must be a first extraction with organic solvent. I supposed you did it on silica column and you eluted with organic solvent. How are you sure it is only fatty acids ?

If you want to get the overall fatty acids concentration, gravimetric methods is the easiest way to estimate total fatty acid from dry weight material.

If you need to know each fatty acid concentrations, you can try FAME analysis by GC-FID with external calibration. There are plenty of litterature about this method on the web. Furthermore, you will check the nature of the fatty acid.

Hope this help,

The question is NOT quite clear.

If you are sure that the extract is only fatty acid !!! take the volume then evaporate the solvent and weigh the fatty acid, the weight to the volume is concentration .Otherwise, follow the procedure described by Leclercq.

Dear Devika Srivastava ,

I suggest you to do an expierement based on FAME. You could use GC to detect your concentration and types of fatty acids compounds in your sample. My paper about it is under review now. If you need, I could send it to you once it published. Hopes it could help you.

Best regards,

Weihao

Hi

The calculation and processing protocols are well explained in the following paper:

https://doi.org/10.1007/s12161-021-02203-0

I hope this helps!

https://www.scielo.br/j/jbchs/a/TxdBvdT4dz7DHSc9Zyv6p3Q/?lang=en

Christie, W.W. Structural analysis of fatty acids. In: Advances in Lipid Methodology - Four, pp. 119-169 (ed. W.W. Christie, Oily Press, Dundee) (1997).

Nikolova-Damyanova, B. Reversed-phase high-performance liquid chromatography: general principles and application to the analysis of fatty acids and triacylglycerols. In: Advances in Lipid Methodology - Four, pp. 193-251 (ed. W.W. Christie, Oily Press, Dundee) (1997).

Good luck.