High-performance liquid chromatographic analysis of amino acids in physiological fluids: on-line precolumn derivatization with o-phthaldialdehyde.
The reaction between primary amines and o-phthaldialdehyde in the presence of a thiol was exploited to measure the concentrations of 21 amino acids in serum by high-performance liquid chromatography with fluorescent detection. The method described here includes an automatic on-line precolumn procedure for derivatizing the amino acids, permitting full automation and avoiding problems due to time-dependent decay of fluorescence of the derivatives. The total analysis time is less than 60 min and limits of sensitivity are about 100 fmol. Proline, hydroxy-proline, and cysteine are not detected. Comparison with data generated by a standard amino acid analyzer shows this technique to be reliable.
D000447:Aldehydes; D000596:Amino Acids / Q000032:analysis* / Q000097:blood; D000818:Animals; D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D009862:Online Systems; D051381:Rats; D009764:o-Phthalaldehyde
D000447:Aldehydes; D000596:Amino Acids; D009764:o-Phthalaldehyde