Post-column fluorescence derivatization of peptides. Problems and potential in high-performance liquid chromatography.
Some  critical parameters such as the pumping system, mixing devices and detector design in instrumentation for post-column derivatization in high-performance liquid chromatography are discussed. The derivatization was studied with pharmaceutically important nona-peptides containing primary amino groups which react with Fluram and reaction parameters such as pH, solvent and reagent concentration were investigated. Both adsorption systems and reversed-phase systems were used to separate the peptides prior to the post-column reaction. Reversed-phase chromatography has the advantage of simpler sample preparation, better reaction control and optimization of solvent conditions. As a result, detection limits of between 5 and 10 ng per injection can be obtained and the reproducibility of the results is better than +/- 2% (relative standard deviation). The method has been applied to the analysis of injection solutions (ampoules).
D000595:Amino Acid Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D008236:Lypressin / Q000032:analysis; D010121:Oxytocin / Q000032:analysis; D010455:Peptides / Q000032:analysis*; D012997:Solvents; D013050:Spectrometry, Fluorescence; D014667:Vasopressins / Q000031:analogs & derivatives
D007202:Indicators and Reagents; D010455:Peptides; D012997:Solvents; D014667:Vasopressins; D010121:Oxytocin; D008236:Lypressin