The LC-MS setup I currently dispose of wouldn't work and changing the column would require a long time so, for now, LC-MS is not an option. HPLC could be an option but the machine I dispose of is also new and still in setup process.
My Gly-Gly dipeptide absorbs light at 224nm, which would be good to detect it but the other component of the initial mixture absorbs at the same wavelenght. That's why I've been looking at the following colorimetric tests:
I have been looking at typical Bradford, Biuret, BCA tests for proteins but as far as I know most tend to work with big proteins only and some only detect a few amino acids (Trp, Cys, Tyr...) but not Gly.
Any ideas?
The link gives a Fluorescence assay for dipeptides using fluorescamine in the presence of amino acids. To separate Gly and diGly I used a Phenomenex Luna silica gel column eluted with 0.1% formic acid in water with UV detection at 210nm.
http://www.sciencedirect.com/science/article/pii/0003269775906909
TLC might be an effective option that doesn't require fancy gear. Detect probably with ninhydrin or 254 fluorescence quenching.
If you have access to an MS system why not just do it by direct infusion - no need for any chromatography. Just look for the parent mass and id you want do MS/MS to check for the right daughters.
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