Optimization of conditions for the simultaneous separation of ten tryptophan metabolites using reversed-phase high-performance liquid chromatography.
A reversed-phase high-performance liquid chromatographic (RP-HPLC) method for the separation of tryptophan and ten metabolites of tryptophan pyrrolase pathway has been developed by sequential optimization of mobile phase, by adjusting the pH, the concentration of triethylamine and the gradient elution. The baseline resolution of the compounds, by this optimized procedure, is obtained with an analysis time, including the re-equilibration period, of less than 30 min. We believe this is the first RP-HPLC method that can separate tryptophan and ten of its metabolites in a single chromatographic run.
Chuang
C Z
CZ
Laboratory of Neurosciences, Pennington Biomedical Research Center, Baton Rouge, LA 70808-4124.
Ragan
F A
FA
Prasad
C
C
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005021:Ethylamines; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D014364:Tryptophan / Q000032:analysis* / Q000378:metabolism
D005021:Ethylamines; D007202:Indicators and Reagents; D014364:Tryptophan; C016162:triethylamine