Is this possible with HPLC? - chech the purity (Apr/10/2006 )

I don't know much about HPLC, so here comes my question.
I've isolated collagen type II but I don't know if it's pure or that there are other collagen types in my sample. Can I check with a HPLC if my sample is pure for this collagen type?

---

HPLC could show you..you'd be looking for a narrow band. Even then though, you don't know for certain that the narrow band doesn't contain other proteins.

You could try running a gel. Find out the molecular mass of your collagen and run your sample with a known pure control (bought from a company).

What detector system are you using with the HPLC? Could you get a UV/Vis spectrogram of your sample and compare it with that from a known pure control. If you have access to a scanning spectrometer you could do th esame without the HPLC step. Or do you use that as part of your purification protocol?

I worked in protein x-ray crystallography and there was a trick of using the unit cell dimensions to check the crystals were from a pure not hybridized sample. If my protein had been twinned I'd have a change in the dimensions. A bit extreme just to check purity.

Good luck.

---