Reverse phase chromatography utilizes the hydrophobic interactions between the column and the amino acids. Therefore, theoretically polar amino acids should be eluted first while more hydrophobic aminocids should be eluted at the end. I want to know why lysine being a basic amino acid is eluted in the end during reverse phase chromatography (HPLC C-18 column, PITC derivitization)?
Lysine has a long aliphatic chain leading up to the epsilon amino group. More importantly, there are two amino groups available for derivatization so you should have a doubly-derivatized amino acid (with twice the signal as well).
If you mention the names of other amino acids which are eluted at the beginning (prior to Lysine) , it is very easy to explain the scenario using the theory mentioned by Michael.
Got it!
Great thanks MIcheal,
You answer was very crisp and to the point.
Dear Arvinna,
all other aminoacids are eluted before lysine using PITC DERIVITIZATION.
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