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- Posts: 2
- Joined: Tue May 22, 2012 4:35 pm
I'm working on an amino acid analysis method. Currently, I'm running a gradient method with a C-18 column:
Column: Atlantis C-18, 100 x 2.1mm, 5µm
MPA: 0.1 M Ammo acetate Buffer
MPB: 100% MeOH
Detection: UV-Vis (338nm)
and I'm preparing the samples by using automated pre-column derivatization with o-phthalaldehyde (OPA). I've only been looking at alanine and glutamine thus far, but I'm hoping to be able to see all the primary and secondary amino acids for future research needs.
I recently read a paper that discusses all the different AA analysis methods, including AA being derivatized with:
1. Ninhydrin (post-column)
2. OPA + Sodium hypochlorite (post-column)
3. PITC (pre-column)
4. AQC (pre-column)
5. DABS-Cl (pre-column)
6. FMOC-Cl (pre-column)
7. NBD-F (pre-column)
It seems to me that maybe the FMOC-Cl is the best method for HPLC with fluorescence detector capability, would you agree?
In the paper, it also discusses the importance of hydrolysis of the samples in preparation for the analysis, would you please share your thoughts on that?
Thank you!!