Pseudomorphic synthesis of bimodal porous silica microspheres for size-exclusion chromatography of small molecules

In this work, mesoporous silica microspheres with bimodal porous structures for size exclusion chromatography (SEC) supports were synthesized via a pseudomorphic transformation method by using 3.5 and 5 μm commercial silica particles as sources and cetyltrimethylammonium bromide (CTAB) as a template. The effects of the synthetic conditions on the pore size distribution were examined, including the temperature, reaction time and the molar ratio of SiO₂:NaOH. Bimodal porous silicas (BPSs) with pore sizes of 3.01 and 12.80 nm were obtained with SiO₂:NaOH:CTAB:H₂O=1:0.1:0.1:20 at 80 °C for 24 h. The BPSs were bonded with diol groups to produce a stationary phase for SEC. The column performance was evaluated with three types of samples, namely, dextran (70 KDa-62 Da), polyethene glycol (PEG) (20 KDa-32 Da) and three biomolecules (36 KDa-1.36 KDa). The column that was packed with a 3.5 μm stationary phase showed excellent resolution for molecular weights of less than 1 KDa with high column efficiency. Carbohydrate samples (dextran (MW=1296), dextran (MW=972), sucrose (MW=342), glucose (MW=180) and glycerol (MW=92)) were separated. Heptaethylene glycol, hexaethylene glycol, pentaethylene glycol, tetraethylene glycol, triethylene glycol, and diethylene glycol were resolved in a PEG200 sample. In summary, this work shows the advantages of bimodal mesopores in SEC for small molecules less than 1 kDa. In the pseudomorphic synthesis, the pore size can be regulated by template micelles. Thus, the development SEC supports with high accuracy for a specified molecular weight range is expected since the pore size can be regulated by the surfactant template.