Analysis of hyaluronic acid by reversed phase HPLC

[archie]

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!

[Vlad Orlovsky]

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.

[unmgvar]

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

[Fernando]

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!

[Vlad Orlovsky]

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

[archie]

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?

[zheyin]

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!

[Vlad Orlovsky]

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.

[max_planck]

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?

[Vlad Orlovsky]

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

[ydna1977]

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

[ydna1977]

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 ?