Hello there!!
I'm analyzing aminoacids by HPLC following the Agilent AAA column method (precolumn derivatization program).
The problem is that the first run that I make it elutes quite good, with a good resolution (figure 1), but in the third run the Rf of the AA changes dramatically (figure 2).
I use a AAA Agilent column (the gradient program is in the figure 3, Solvent C is ACN:MeOH:H2O 45:45:10, solvent D is Na2HPO4 40mM pH 7.8).
I would like you to help me to identify what could be happening.
Thank you very much!!
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