Novel monolithic poly(phenyl acrylate- co-1,4-phenylene diacrylate) capillary columns for biopolymer chromatography

• Published in: Journal of Chromatography A 2007-04, Vol.1147 (1), p.46-52
• Main Authors: Bisjak, Clemens P., Lubbad, Said H., Trojer, Lukas, Bonn, Guenther K.
• Format: Article
• Language: English
• Subjects: 1,4-Phenylene diacrylate (PDA); Acrylates - chemical synthesis; Acrylates - chemistry; Analytical biochemistry: general aspects, technics, instrumentation; Analytical chemistry; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Biopolymers - isolation & purification; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - classification; Exact sciences and technology; Free Radicals - chemistry; Fundamental and applied biological sciences. Psychology; Micro-HPLC; Microscopy, Electron, Scanning; Molecular Structure; Monolithic capillary columns; Nucleic Acids - isolation & purification; Oligodeoxynucleotides; Other chromatographic methods; Permeability; Phenyl acrylate (PA); Polymers - chemical synthesis; Polymers - chemistry; Porosity; Proteins; Proteins - isolation & purification; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2007.02.027

Monolithic capillary columns were prepared by thermally initiated free radical polymerisation of phenyl acrylate (PA) and 1,4-phenylene diacrylate (PDA) in the confines of 200 μm I.D. fused silica capillaries. Polymerisation was performed in the presence of 2-propanol and tetrahydrofuran (THF) as inert diluents (porogens), using α,α′-azoisobutyronitrile (AIBN) as initiator. Morphology and porosity of the resulting monoliths were comprehensively studied by scanning electron microscopy (SEM), mercury intrusion porosimetry and inverse size-exclusion chromatography (ISEC). The novel poly(phenyl acrylate- co-1,4-phenylene diacrylate) (PA/PDA) monoliths showed high mechanical stability and were successfully applied to the separation of proteins and oligodeoxynucleotides, employing reversed-phase (RP) and ion-pair reversed-phase (IP-RP) conditions, respectively. Maximum loading capacities for cytochrome c and d(pT) 16 were evaluated and found to be in the region of 200 fmol. Batch-to-batch reproducibility was determined for three independently prepared PA/PDA monolithic capillary columns. Relative standard deviations (RSDs) of retention time ( t R) of 0.7–1.6% for proteins and 0.2–2.5% for d(pT) 12–18 proved high reproducibility of the PA/PDA supports.