Comparison between monolithic conventional size, microbore and capillary poly(p -methylstyrene -co -1,2 -bis(p -vinylphenyl )ethane) high-performance liquid chromatography columns synthesis, application, long-term stability and reproducibility
Novel monolithic supports (MS/BVPE) were prepared by thermally initiated free radical copolymerisation of p-methylstyrene (MS) and 1,2-bis(p-vinylphenyl)ethane (BVPE). The polymer was synthesised in fused silica capillaries (80 mm x 0.2 mm and 80 mm x 0.53 mm) and in borosilicate glass columns (90 m...
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Published in | Journal of chromatography Vol. 1146; no. 2; pp. 216 - 224 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Amsterdam
Elsevier
06.04.2007
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Subjects | |
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Abstract | Novel monolithic supports (MS/BVPE) were prepared by thermally initiated free radical copolymerisation of p-methylstyrene (MS) and 1,2-bis(p-vinylphenyl)ethane (BVPE). The polymer was synthesised in fused silica capillaries (80 mm x 0.2 mm and 80 mm x 0.53 mm) and in borosilicate glass columns (90 mm x 1.0 mm and 90 mm x 3.0 mm) to yield different HPLC column designs. A comparison of those column dimensions regarding morphology as well as separation efficiency and applicability in bioanalysis is presented. The efficiency towards proteins as well as oligonucleotides was found to be considerably improved with decreasing column I.D. While a 5-protein mixture was baseline separated on all investigated column designs, the separation of small biomolecules like oligonucleotides or peptides on microbore and conventional size glass columns was strongly restricted in terms of resolution due to extensive peak broadening or the occurrence of peak asymmetry. Monolithic MS/BVPE capillary columns up to 0.53 mm I.D., however, proved to be applicable to the fractionation of the whole spectrum of biopolymers, including proteins, peptides, oligonucleotides as well as double-stranded DNA fragments. Due to the fact that reliable chromatography makes great demand on the robustness of the stationary phase, monolithic MS/BVPE capillaries were subjected to a comprehensive reproducibility study including run-to-run as well as batch-to-batch reproducibility. |
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AbstractList | Novel monolithic supports (MS/BVPE) were prepared by thermally initiated free radical copolymerisation of p-methylstyrene (MS) and 1,2-bis(p-vinylphenyl)ethane (BVPE). The polymer was synthesised in fused silica capillaries (80 mm x 0.2 mm and 80 mm x 0.53 mm) and in borosilicate glass columns (90 mm x 1.0 mm and 90 mm x 3.0 mm) to yield different HPLC column designs. A comparison of those column dimensions regarding morphology as well as separation efficiency and applicability in bioanalysis is presented. The efficiency towards proteins as well as oligonucleotides was found to be considerably improved with decreasing column I.D. While a 5-protein mixture was baseline separated on all investigated column designs, the separation of small biomolecules like oligonucleotides or peptides on microbore and conventional size glass columns was strongly restricted in terms of resolution due to extensive peak broadening or the occurrence of peak asymmetry. Monolithic MS/BVPE capillary columns up to 0.53 mm I.D., however, proved to be applicable to the fractionation of the whole spectrum of biopolymers, including proteins, peptides, oligonucleotides as well as double-stranded DNA fragments. Due to the fact that reliable chromatography makes great demand on the robustness of the stationary phase, monolithic MS/BVPE capillaries were subjected to a comprehensive reproducibility study including run-to-run as well as batch-to-batch reproducibility. |
Author | TROJER, Lukas BONN, Günther K WIEDER, Wolfgang LUBBAD, Said H BISJAK, Clemens P |
Author_xml | – sequence: 1 givenname: Lukas surname: TROJER fullname: TROJER, Lukas organization: Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, Innrain 52a, 6020 Innsbruck, Austria – sequence: 2 givenname: Said H surname: LUBBAD fullname: LUBBAD, Said H organization: Department of Chemistry, Alazhar University-Gaza, P.O. Box 1277, Gaza, Israel – sequence: 3 givenname: Clemens P surname: BISJAK fullname: BISJAK, Clemens P organization: Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, Innrain 52a, 6020 Innsbruck, Austria – sequence: 4 givenname: Wolfgang surname: WIEDER fullname: WIEDER, Wolfgang organization: Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, Innrain 52a, 6020 Innsbruck, Austria – sequence: 5 givenname: Günther K surname: BONN fullname: BONN, Günther K organization: Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University of Innsbruck, Innrain 52a, 6020 Innsbruck, Austria |
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Keywords | Capillary column Peptides Chromatographic retention Oligonucleotides HPLC chromatography High-performance liquid chromatography (HPLC) Styrene derivative copolymer Proteins Separation method Double stranded DNA Oligonucleotide p-Methylstyrene (MS) Stationary phase Stability Monolith Monolithic column Long term Protein Microbore column Analysis method l,2-bis(p-Vinylphenyl)ethane (BVPE) Reproducibility Preparation Comparative study dsDNA |
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SubjectTerms | Analytical, structural and metabolic biochemistry Biological and medical sciences Chromatography, High Pressure Liquid - instrumentation Chromatography, High Pressure Liquid - methods DNA - chemistry Dna, deoxyribonucleoproteins Fundamental and applied biological sciences. Psychology General aspects, investigation methods Nucleic acids Oligonucleotides - chemistry Peptides - chemistry Proteins Proteins - chemistry Reproducibility of Results Silicon Dioxide - chemistry Styrene - chemistry Styrenes - chemistry Vinyl Compounds - chemistry |
Title | Comparison between monolithic conventional size, microbore and capillary poly(p -methylstyrene -co -1,2 -bis(p -vinylphenyl )ethane) high-performance liquid chromatography columns synthesis, application, long-term stability and reproducibility |
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