Abstract
•An amide embedded C18 stationary phase based on silica monolith particles was prepared.•Peptides, proteins, polycyclic aromatic hydrocarbons, and nucleotides were separated with good separation efficiency.•The performance of C18-amide column was better than commercial amide columns.
Peptides and proteins have great therapeutic potential, and owing to their unique biological properties, their separation is very challenging. A polar embedded C18 stationary phase was prepared by functionalization of porous silica monolith particles with N-3-(trimethoxysilyl) propyl stearamide. The stationary phase was packed in narrow bore column (100 × 2.1 mm) and evaluated for the separation of peptides and proteins in HPLC. Number of theoretical plates (N) as high as 520,000 and 340,000 plates/meter were obtained for peptides and proteins respectively. The separation performance of C18-amide column was compared with four commercial amide columns i.e. Ascentis Express RP-Amide, Accucore 150 Amide, TSKgel Amide-80, and XBridge Amide column. The performance of C18-amide column was much better than these commercial amide columns in terms of efficiency and resolution. Owing to high efficiency and good resolution for peptides and proteins separation, C18-amide column could be used as alternative to expansive commercial columns.