Chromatography Forum

Hi All,

I am trying to detect the presence of glutamic acid in my protein hydrolysate.

Has anyone tried this before using an HPLC method?

If so, can you please recommend: -
Column:
Eluent:
Flow rate:
Injection volume:
Detection (nm):

What retention time was obtained?

Regards

Gavin

Column: Sumichiral OA-5000 Column 150 x 4.6 mm
Part No:CH0-1859 from Phenomenex
Dimensions:150 x 4,6 mm ID
Elution Type:Isocratic
Elution A:2 mM Copper (II) sulfate in water
Elution B:Methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 85 15


Flow Rate:1 mL/min
Col. Temp: ambient Detection: UV-Vis Abs. - Variable Wave. @ 254 nm (22°C)

SIELC has also some nice applications, and other manufactureres as well. Good luck

Is it not possible to use a C18 column and only one mobile phase?

Gavin

Application Note SI-01214 Effective Determination of Glutamic Acid ... - [ Diese Seite übersetzen ]Dateiformat: PDF/Adobe Acrobat - Schnellansicht
HPLC is often used in assays for the determination of glutamic acid. In this example, an isocratic HPLC method was employed to analyze ...
www.chem.agilent.com/Library/applications/SI-01214.pdf
The chromatography conditions might be a good starting point. I would recommend to do SPE before going to HPLC, to remove all proteins in your sample. Good luck.

Check out the first publication at the following post:

http://chromforum.org/viewtopic.php?f=1&t=6509

Choose the one you like:
http://www.sielc.com/Compound-Glutamic-Acid.html

Please see if the following information helps:

1. http://www.dionex.com/en-us/images/pdf- ... e-beer.pdf
2. http://www.dionex.com/en-us/images/pdf- ... antage.pdf