Novel monolithic poly(phenyl acrylate-co-1,4-phenylene diacrylate) capillary columns for biopolymer chromatography.
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 microm I.D. fused silica capillaries. Polymerisation was performed in the presence of 2-propanol and tetrahydrofuran (THF) as inert diluents (porogens), using alpha,alpha'-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.
Bisjak
Clemens P
CP
Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 52a, 6020 Innsbruck, Austria.
Lubbad
Said H
SH
Trojer
Lukas
L
Bonn
Guenther K
GK
D000179:Acrylates / Q000138:chemical synthesis* / Q000737:chemistry; D001704:Biopolymers / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000145:classification*; D005609:Free Radicals / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D015394:Molecular Structure; D009696:Nucleic Acids / Q000302:isolation & purification; D010539:Permeability; D011108:Polymers / Q000138:chemical synthesis* / Q000737:chemistry; D016062:Porosity; D011506:Proteins / Q000302:isolation & purification; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D000179:Acrylates; D001704:Biopolymers; D005609:Free Radicals; D009696:Nucleic Acids; D011108:Polymers; D011506:Proteins; C521539:poly(phenyl acrylate-co-1,4-phenylene diacrylate)