Chromatography Forum

Posted: **Tue Oct 10, 2006 10:22 am**

Hi everyone,

could someone advice me a suitable method for analysis of betaine (HPLC or other methods)?
i'm trying an UV method with reinecke salt but validation is quiet plodding and difficult...
thank you in advance
vincenzo

Posted: **Tue Oct 10, 2006 12:09 pm**

Here is method for betaine. You have better chances with ELSD/LC/MS detection. For low UV you can use sulfuric or phosphoric acid (but betaine has limited UV activity)

http://www.sielc.com/compound_197.html

This analysis is performed on 4.6x50 mm column. If you have other compounds in the mixture you might consider a longer column (100 or 150 mm)

Posted: **Tue Oct 10, 2006 1:03 pm**

Hey Vincenzo -

Here is a chromatogram of betaine using our Unison UK-Silica column:

http://www.imtakt.com/TecInfo/TI191E.pdf

Posted: **Tue Oct 10, 2006 5:43 pm**

I would use HILIC on a silica column. Roughly 70 to 80% acetonitrile, 30 to 20 % water.

Posted: **Tue Oct 10, 2006 8:40 pm**

See Polymer Laboratories Application Note:

TB1074: Quantification of Betaine in Chinese Lycium using Hydrophilic Interaction Chromatography (HILIC) and ELS Detection.

http://www.polymerlabs.com/cgi-bin/fill ... chbull.htm

Do I need to say that they use ZICÂ®-HILIC...

Posted: **Thu Oct 12, 2006 4:22 pm**

There is a very simple method available using ligand exchange columns:

http://chromatography.transgenomic.com/ ... .asp?tab=2

Bart Poulsen

Transgenomic, Inc.
Director, Chromatography Product Management

Posted: **Thu Oct 12, 2006 5:18 pm**

One of my colleagues told me my link did not work. It will take a bit longer to get there, but just in case, you can start at our home web site and find it after some searching: chromatography/carbohydrate analysis/Coregel 87K/chromatogram.

www.transgenomic.com

Bart

Posted: **Fri Oct 13, 2006 1:14 pm**

a question:

is possible in your opinion to use a conventional c18 column at low pH values (2.1-2.

and a ion pair reagent like eptanSO4?at theese pH betaine should be a cation so if i prepare mobile phase with ion pair reagent it could be ok...
what about??
thank you

Posted: **Wed Oct 18, 2006 9:25 pm**

There may be a possibility that you can suppress the carboxylic acid group and retain it as a ion pair with heptan sulfonate on a RP column.

However, I suspect that you will have difficulties with the detection since UV absorbtion by betaine is rather low and the ion-pairing is not compatible with ELSD or MS detection.