Chromatography Forum

I have extracted a compound from nature product and I want to use this compound as standard for developing a analytical method using HPLC.

The question is how could I know the exact purity of this compound if it does not has any commercial working or reference standard.

You will never get an exact purity, but you can probably get close enough for your purpose via NMR, MS, UV, IR, and elemental (carbon, hydrogen , , , ) analysis. Two very different chromatographic methods may also help, sometimes a sharp (constant after several recrystallizations) melting point is quite instructive.

Moisture content would also be helpful.