I am looking for a simple HPLC method for separation of biological amino acids and their oligomers. I have access to HPLC with C18 column and UV detection system. I can find lots of fluorescent methods and MS analysis methods but do not have access to necessary equipment. I would prefer a pre-column derivitisation if possible. Any advice, help or ideas would be greatly received, thanks.
For amino acid determination you could use the pico tag method. In this method you use PTIC for the pre-column derivatization of amino acids. The best resolution is the obtained with C18 (250 x 4.6 x 5 um) column and the detection using an UV detector at 254 nm.
Regards.
We also use fluoresent method, it gives very good seperation.
Yes, we also use the fluorescent method (AccQ-Tag), and I prefer it because is faster and easier than pico-tag derivatization and the hplc resolution is so well. However, in my opinion, (and Barry said that he only have access to an uv detector) pico-tag coul be the best alternative if you want to use PDA or UV detector.
Hi,
you may want to check the answers found here:
https://www.researchgate.net/post/How_can_I_analyze_amino_acids_with_HPLC-UV
and work by Gómez-Alonso, Hermosín-Gutiérrez, and García-Romero (2007) Journal of Agricultural and Food Chemistry, 55, 608-613.
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