Peptide content-determination of crude synthetic peptides by reversed-phase liquid chromatography and nitrogen-specific detection with a chemiluminescent nitrogen detector

The need for a rapid and accurate method to assess peptide content is driven by the recent surge in demand for pharmaceutical-grade synthetic peptides manufactured by solid-phase chemistry. Routinely, the purity of synthetic peptides is analyzed by reversed-phase HPLC with UV detection, and peptide content is typically determined from amino acid analysis of synthetic peptides. Recently, we developed a new method to assess the peptide content of synthetic peptides in one simple technique: simultaneous reversed-phase HPLC-chemiluminescent nitrogen detection and UV detection. In this paper we present the simplicity of this method and its universal application to analyzing crude synthetic peptides produced by solid-phase synthesis, an important first step in manufacturing bioactive peptides.