Research Article
BibTex RIS Cite

Design of Reversed Phase Chromatography (RPC) Column Packing Material for Protein Separation: Part 1- Synthesis and Characterization

Year 2017, Volume: 38 Issue: 4, 788 - 797, 08.12.2017
https://doi.org/10.17776/csj.349325

Abstract

In the first part of the two-part study, the column packed
materials in the form of monodisperse and porous particles were synthesized
with regard to test their column performances in Reverse Phase Chromatography
(RPC). Particles in the form of poly(vinyl acetate-co-divinylbenzene)
[poly(VAc-co-DVB)] and poly(divinylbenzene) [poly(DVB)] were produced by
“multi-stage micro-suspension polymerization”. The surface of particles were
converted to “hydroxyl” form by base hydrolysis that were used as
poly(vinylalcohol-co-divinylbenzene) [poly(VA-co-DVB)] particles, having
appropriate structural properties like surface and porosity for chromatographic
applications. Particle characterization was assessed by Scanning Electron
Microscopy (SEM), BET surface area and porosity measurement system and Fourier
Transform Infrared Spectroscopy (FTIR).

References

  • [1]. Camlı S.T., Şenel S., Tuncel A., Cibacron Blue F3G-A Attached Uniform and Macroporous Poly(styrene-divinlybenzene) Particles For Specific Albumin Adsorption. J. Biomater. Sci. Polym., 1999, 8, 875-889.
  • [2]. Ellingsten T., Aune O., Ugelstad J., Hagen S., Mono-sized Stationary Phases for Chromatography, Journal of Chromatography A, 1990, 535, 147-161.
  • [3]. Galia M., Svec F., Fréchet J.M.J., Monodisperse Polymer Beads as Packing Material for High-performance Liquid Chromatography. Effect of Divinylbenzene Content on the Porous and Chromatographic Properties of Poly(styrene-co-divinylbenzene) Beads Prepared in Presence of Linear Polystyrene as a Porogen, J. Polym. Sci., Polym. Chem., 1994, 32, 2169-2175
  • [4]. Hossoya K., Teramachi M., Tanaka N., Kobayashi A., Kanda T., Ohtsu Y., Preparation Strategy for Uniformly Sized, Polymer-Beads HPLC Packing Materials Having Practically Acceptable Column Efficiency. 1. Copolymerization Technique, Anal. Chem., 2001, 73(24), 5852-5857
  • [5]. Lewandowski K., Svec F., Frechet J.M.J., Polar, A Novel Polar Seperation Medium For The Size Exclusion Chromatography of Small Molecules: Uniformly Sized, Porous Poly(vinylphenol-co-divinylbenzene) Beads, J. Liq. Chrom. & Rel. Technol., 1997, 20, 227-243
  • [6]. Smigol V., Svec F., Frechet J.M.J., 2-Dimensional High-Performance Liquid-Chromatography Using Monodisperse Polymer Beads Containing Segregated Chemistries Prepared By Pore-Size Specific Functionalization-Single-Column Combinations of Size Exclusion Or İon-Exchange With Reversed-Phase Chromatography, Analytical Chemistry, 1994, 66 (23), 4308-4315
  • [7]. Smigol V., Svec F., Frechet J.M.J., High-Performance Liquid Chromatography of Complex mixtures Using Monodisperse Dual- Chemistry Polymer Beads Prepared by a Pore-Size-Specific Functionalization Process. A Single Column Combination of Hydrophobic Interaction and Reversed-Phase Chromatography, Analytical Chemistry, 1994, 66(13), 2129-2138
  • [8]. Unsal E., Durdu A., Elmas B., Tuncel M., Tuncel A., A New Affinity-HPLC Packing For Protein Separation: Cibacron Blue Attached Uniform-Porous Poly(HEMA-co-EDM) Beads, Analytical and Bioanalytical Chemistry, 2005, 383, 930-937
  • [9]. Unsal E., Irmak T., Durusoy E., Tuncel M., Tuncel, A., Monodisperse porous particles with polyionic ligands for ion exchange separation of proteins, Analytical Chimica Acta, 2006, 570, 240-248
  • [10]. Ettre L.S., Chromatography: the Separation Technique of the 20th Century. Chromatographia, 2000, 51 (1/2), 7-17
  • [11]. Engelhardt H., One century of liquid chromatography from Tswett’s columns to modern high speed and high performance separations, Journal of Chromatography B, 2004, 800(1-2), 3-6
  • [12]. Zhang Y.W., Yao S., Zeng H., Song H., Chiral Separation of Pharmaceuticals by High Performance Liquid Chromatography, Current Pharmaceutıcal Analysıs, 2010, 6 (2), 114-130
  • [13]. Unsal E., Elmas B., Camli S.T., Tuncel M., Senel S., Tuncel A., Monodisperse-Porous Poly(Styrene-co-Divinylbenzene) Beads Providing High Column Efficiency in Reversed Phase HPLC, Journal of Applied Polymer Science, 2005, 95: 1430-1438
  • [14]. Unsal E., Camli S.T., Irmak T., Tuncel M., Tuncel A., Monodisperse poly(styrene-co-divinylbenzene) particles (3.2 μm) with relatively small pore size as HPLC packing material, Chromatographia, 2004, 60: 553-560
  • [15]. Unsal E., Camli S.T., Senel S., Tuncel A., Chromatographic Performance of Monodisperse–Macroporous Particles Produced by “Modified Seeded Polymerization.” I: Effect of Monomer/Seed Latex Ratio, Journal of Applied Polymer Science, 2004, 92: 607-618
  • [16]. Unsal E., Elmas B., Camli S.T., Senel S., Tuncel A., A Uniform Particles For The Reversed Phase Separation of Proteins With High Resolution and High Column Efficiency, Separation Science and Technology, 2005, 40: 1425-1438
  • [17]. Çağlayan B., Unsal E., Çamlı T., Tuncel M., Tuncel A., Monodisperse Porous Poly(Vinyl Acetate-Codivinylbenzene) Particles By Single-Stage Seeded Polymerization: A Packing Material For Reversed Phase HPLC. J. Sep. Sci., 2006, 29: 936-944

Proteinleri Ayırmak İçin Ters Faz Kromatografisi (RPC) Kolon Dolgu Materyali Tasarımı: Bölüm 1- Sentez ve Karakterizasyon

Year 2017, Volume: 38 Issue: 4, 788 - 797, 08.12.2017
https://doi.org/10.17776/csj.349325

Abstract

İki bölümden oluşan çalışmanın ilk bölümünde
Ters Faz Kromatografisi (Reversed Phase Chromatography, RPC)
için eş boyutlu ve
gözenekli partikül formunda poli(vinil asetat-co-divinilbenzen)
[poli(VAc-co-DVB)] ve poli(divinilbenzen) [poli(DVB)] kolon dolgu materyalleri
“Çok Basamaklı Mikrosüspansiyon Polimerizasyonu” yöntemi ile sentezlenmiştir.
Partiküller bazik ortamda hidrolizlenerek, partikül yüzeyi “hidroksil” formuna
dönüştürülmüş ve kromatografik kullanım için uygun yüzey ve gözenek yapısına
sahip poli(vinilalkol-co-divinilbenzen) [poli(VA-co-DVB)] partiküller elde
edilmiştir. Partiküllerin karakterizasyonu Taramalı Elektron Mikroskobu
(Scanning Electron Microscopy, SEM), yüzey alanı ve gözeneklilik ölçüm sistemi
(Brunauer–Emmett–Teller, BET) ve Fourier Transfer Infrared Spektrofotometre
(Fourier Transform Infrared Spectroscopy, FTIR) ile yapılmıştır.

References

  • [1]. Camlı S.T., Şenel S., Tuncel A., Cibacron Blue F3G-A Attached Uniform and Macroporous Poly(styrene-divinlybenzene) Particles For Specific Albumin Adsorption. J. Biomater. Sci. Polym., 1999, 8, 875-889.
  • [2]. Ellingsten T., Aune O., Ugelstad J., Hagen S., Mono-sized Stationary Phases for Chromatography, Journal of Chromatography A, 1990, 535, 147-161.
  • [3]. Galia M., Svec F., Fréchet J.M.J., Monodisperse Polymer Beads as Packing Material for High-performance Liquid Chromatography. Effect of Divinylbenzene Content on the Porous and Chromatographic Properties of Poly(styrene-co-divinylbenzene) Beads Prepared in Presence of Linear Polystyrene as a Porogen, J. Polym. Sci., Polym. Chem., 1994, 32, 2169-2175
  • [4]. Hossoya K., Teramachi M., Tanaka N., Kobayashi A., Kanda T., Ohtsu Y., Preparation Strategy for Uniformly Sized, Polymer-Beads HPLC Packing Materials Having Practically Acceptable Column Efficiency. 1. Copolymerization Technique, Anal. Chem., 2001, 73(24), 5852-5857
  • [5]. Lewandowski K., Svec F., Frechet J.M.J., Polar, A Novel Polar Seperation Medium For The Size Exclusion Chromatography of Small Molecules: Uniformly Sized, Porous Poly(vinylphenol-co-divinylbenzene) Beads, J. Liq. Chrom. & Rel. Technol., 1997, 20, 227-243
  • [6]. Smigol V., Svec F., Frechet J.M.J., 2-Dimensional High-Performance Liquid-Chromatography Using Monodisperse Polymer Beads Containing Segregated Chemistries Prepared By Pore-Size Specific Functionalization-Single-Column Combinations of Size Exclusion Or İon-Exchange With Reversed-Phase Chromatography, Analytical Chemistry, 1994, 66 (23), 4308-4315
  • [7]. Smigol V., Svec F., Frechet J.M.J., High-Performance Liquid Chromatography of Complex mixtures Using Monodisperse Dual- Chemistry Polymer Beads Prepared by a Pore-Size-Specific Functionalization Process. A Single Column Combination of Hydrophobic Interaction and Reversed-Phase Chromatography, Analytical Chemistry, 1994, 66(13), 2129-2138
  • [8]. Unsal E., Durdu A., Elmas B., Tuncel M., Tuncel A., A New Affinity-HPLC Packing For Protein Separation: Cibacron Blue Attached Uniform-Porous Poly(HEMA-co-EDM) Beads, Analytical and Bioanalytical Chemistry, 2005, 383, 930-937
  • [9]. Unsal E., Irmak T., Durusoy E., Tuncel M., Tuncel, A., Monodisperse porous particles with polyionic ligands for ion exchange separation of proteins, Analytical Chimica Acta, 2006, 570, 240-248
  • [10]. Ettre L.S., Chromatography: the Separation Technique of the 20th Century. Chromatographia, 2000, 51 (1/2), 7-17
  • [11]. Engelhardt H., One century of liquid chromatography from Tswett’s columns to modern high speed and high performance separations, Journal of Chromatography B, 2004, 800(1-2), 3-6
  • [12]. Zhang Y.W., Yao S., Zeng H., Song H., Chiral Separation of Pharmaceuticals by High Performance Liquid Chromatography, Current Pharmaceutıcal Analysıs, 2010, 6 (2), 114-130
  • [13]. Unsal E., Elmas B., Camli S.T., Tuncel M., Senel S., Tuncel A., Monodisperse-Porous Poly(Styrene-co-Divinylbenzene) Beads Providing High Column Efficiency in Reversed Phase HPLC, Journal of Applied Polymer Science, 2005, 95: 1430-1438
  • [14]. Unsal E., Camli S.T., Irmak T., Tuncel M., Tuncel A., Monodisperse poly(styrene-co-divinylbenzene) particles (3.2 μm) with relatively small pore size as HPLC packing material, Chromatographia, 2004, 60: 553-560
  • [15]. Unsal E., Camli S.T., Senel S., Tuncel A., Chromatographic Performance of Monodisperse–Macroporous Particles Produced by “Modified Seeded Polymerization.” I: Effect of Monomer/Seed Latex Ratio, Journal of Applied Polymer Science, 2004, 92: 607-618
  • [16]. Unsal E., Elmas B., Camli S.T., Senel S., Tuncel A., A Uniform Particles For The Reversed Phase Separation of Proteins With High Resolution and High Column Efficiency, Separation Science and Technology, 2005, 40: 1425-1438
  • [17]. Çağlayan B., Unsal E., Çamlı T., Tuncel M., Tuncel A., Monodisperse Porous Poly(Vinyl Acetate-Codivinylbenzene) Particles By Single-Stage Seeded Polymerization: A Packing Material For Reversed Phase HPLC. J. Sep. Sci., 2006, 29: 936-944
There are 17 citations in total.

Details

Journal Section Engineering Sciences
Authors

Berna Saraçoğlu Kaya

Ebru Sağ

Publication Date December 8, 2017
Submission Date April 7, 2017
Acceptance Date May 23, 2017
Published in Issue Year 2017Volume: 38 Issue: 4

Cite

APA Saraçoğlu Kaya, B., & Sağ, E. (2017). Design of Reversed Phase Chromatography (RPC) Column Packing Material for Protein Separation: Part 1- Synthesis and Characterization. Cumhuriyet Science Journal, 38(4), 788-797. https://doi.org/10.17776/csj.349325