Chromatography Forum

Hi all..
I am analysing hyaluronic acid in cosmetics.
I found many article that using GPC for that, but I can't afford this column.
So, is it possible to do it in usual RP column? I mean C-18 or C-8.. with 100-120A pore size.
The MW is about 1,400,000 Da.
I don't have wide pore column either.

Thanks in advance!

we have a method for something similar (alginic acid), although it is smaller acidic polymer than you molecule:
http://www.sielc.com/Compound-Alginic-Acid.html

because of the polymeric nature of the molecule, in order to get a better peak you need to go into ion-exclusion/size exclusion mode. Please note that flow rate in our application is 4 times slower.

no you probably cannot do this compound on a standard RP column

you can maybe do an application with a wider pore column, like 300 or maybe better to try a 500 or 1000A pore volume.
even if you see something you'll probably have a very bad recovery on such a column

Archie

I´m sorry you have as all mortals go to the BP 2012 for example, and do it colorimetrically, especially if its a problem of money.

Good luck!

You can do it on smaller pore size columns in size exclusion mode. I think that even 300-500A might be too narrow for this polymer

Ow..
I also find application that using ion pair reagent, but hyaluronic acid need to be derivatized first.
I don't have any experience in derivatizing sample either..
But I think it's rather complicated..am I right?

Ow..
I also find application that using ion pair reagent, but hyaluronic acid need to be derivatized first.
I don't have any experience in derivatizing sample either..
But I think it's rather complicated..am I right?

Derivatization is like wet chemistry,
not complex at all...
just tedious!

What kind of derivatization they are talking about? It is hard to derivatize this polymer completely and you will end up with a horrible mixture of things. Go with size/charge exclusion at low flow rate and low UV, or use ELSD for detection.

we have a method for something similar (alginic acid), although it is smaller acidic polymer than you molecule:
http://www.sielc.com/Compound-Alginic-Acid.html

because of the polymeric nature of the molecule, in order to get a better peak you need to go into ion-exclusion/size exclusion mode. Please note that flow rate in our application is 4 times slower.

Is this working with a UV-detector at 210 nm for hyaluronic acid?

It is uv active Since it has a lot of carbonyls.

Hi ,
On the subject of Hyaluronic acid , what is the most suitable sample extraction technique to achieve suitable sample recovery 90-110% ie shake , vortex, stir , temp..? ..for various MW of HA for example 3-4 Mda in sample material/product

Also what are the options for a force deg study on Hyaluronic acid with respect to ICH Q2 (R1) IE acid base oxidation light thermal etc and suitable conditions ?