Frei R W, Michel L, Santi W
J Chromatogr. 1976 Nov 3;126:665-77. doi: 10.1016/s0021-9673(01)84110-8.
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).
