Amino acid analysis by high-performance liquid chromatography after derivatization with 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide.
Amino acids are quantitatively determined by precolumn derivatization with 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide and reversed-phase high-performance liquid chromatography with photometric detection at 340 nm. Excellent chromatographic resolution of a mixture of the derivatives of 20 amino acids including proline and cystine is achieved within 110 min by linear gradient elution with acetonitrile in 13 mM trifluoroacetate plus 4% (by vol) tetrahydrofuran. The limit of detection is 50 pmol. Amino acid analyses of acid hydrolysates of the several proteins gave results equivalent to those obtained by conventional ion-exchange-based amino acid analysis. The simplicity of the procedure allows its use on any multipurpose high-performance liquid chromatographic system.
Kochhar
S
S
Biochemisches Institut der Universität Zürich, Switzerland.
Christen
P
P
D000409:Alanine / Q000031:analogs & derivatives*; D000596:Amino Acids / Q000032:analysis*; D001219:Aspartate Aminotransferases / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004136:Dinitrobenzenes; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis; D009113:Muramidase / Q000032:analysis; D009578:Nitrobenzenes
D000596:Amino Acids; D004136:Dinitrobenzenes; D009578:Nitrobenzenes; C047134:Marfey's reagent; D001219:Aspartate Aminotransferases; D009113:Muramidase; D005634:Fructose-Bisphosphate Aldolase; D000409:Alanine