Evaluation of selectivity for l-glutamide-derived highly ordered assemblies in reversed-phase high-performance liquid chromatography

Two dioctadecyl l-glutamic acid derivatives with amide and ester type bondings have been synthesized and immobilized from 3-aminopropyltrimethoxysilane (APS) grafted silica (Sil-APS) to be used in reversed-phase high-performance liquid chromatography (RP-HPLC). Subsequent studies showed that dioctadecyl-l-glutamide derivative (GLN) can self-assemble into highly ordered structures by forming three-dimensional fibrillar aggregates as observed in scanning and transmission electron microscopes (SEM and TEM). Variable temperature 1H NMR and FT-IR spectra of organogel revealed that the special aggregation morphology shown by GLN was stabilized by inter and or intra molecular hydrogen bonding among amide moieties. However, such ordered aggregated or self-assembled structures were not observed for the dioctadecyl-l-glutamate (GLU) derivative. The stationary phases Sil-GLN and Sil-GLU were characterized by DRIFT, elemental analysis, TGA, and 13C and 29Si CP-MAS NMR spectroscopic measurements. The chromatographic selectivity for both stationary phases was evaluated from the retention studies of different size and shape polycyclic aromatic hydrocarbons (PAHs). The chromatographic experiment for PAHs and geometrical isomers in RP-HPLC showed that Sil-GLN demonstrated extremely enhanced selectivity than Sil-GLU. The higher selectivity attributed by Sil-GLN has been brought by multiple π–π interactions among the π-electrons of the grafted organic phase and π-electrons of the guest PAHs molecules. Thermodynamic studies for linear and nonlinear PAHs revealed that the retention behavior does not change over a temperature range from 10 to 60 °C for both stationary phases.