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Derivatization of amino acids method

PostPosted: Wed 15 Jun 2016 10:24
by Mickey&Minnie
Hello everybody,

Could you please tell me a method for the derivarization of amino acids?

Thanks,
Mickey ;)

Re: Derivatization of amino acids method

PostPosted: Wed 15 Jun 2016 11:49
by Sean
Hi Mickey,

There are numerous procedures for derivatization of amino acids. I will list a couple :
1) Reagent: Methanol (MeOH)/HCl, TFAA, derivative type methyl ester
Dissolve 2 mg of amino acids in 2 mg of dry MeOH and bubble in dry HCl gas for 30 minutes while heating at 70C. Evaporate to dryness under vac. Add 0.2 ml TFAA and allow to stand at RT for at least 30 minutes. Evaporate under vac while cooling tube in ice bath (limit to 4 min to minimize loss of volatile derivs). Diss in 100 microliters CH2Cl2 and analyze by GC.
2) Reagent : Ethanol, HFBA/Pyridine, compound type: ethyl ester, heptafluorobutyryl)
Dissolve extracted amino acid in 0.1 ml CHCl3. Add 0.01 ml EtOH/CHCl3 (1:4) and mix by shaking. Add 0.01 ml pyridine/CHCl3 (1:3,v/v) and mix by shaking. Add 0.03 ml HFBA and mix bu shaking. Evaporate CHCl3 to ca 1/2 drop and heatin boiling H2O bath 4 minutes. Cool under tap H2O and dill with 0.14 ml CHCl3. Add ca 0.09 ml of 1N HCl and separateChCl3 layer. Wash with 0.1N NaOH and allow to stand for 30 minutes. Separate CHCl3 layer and evaporate solvent almost to dryness. Add 0.1 ml Et2O or acetone for GC analysis.

Have a good day,
Sean

Re: Derivatization of amino acids method

PostPosted: Thu 16 Jun 2016 09:29
by Mickey&Minnie
Thank you Sean!

Mickey ;)