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Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance

Year 2019, , 213 - 220, 22.03.2019

Berna Saraçoğlu Kaya , Ebru Sağ

https://doi.org/10.17776/csj.364625

Abstract

In the second part of the two-part study, monodisperse and porous
poly(vinyl acetate-co-divinylbenzene) [poly(VAc-co-DVB)] and
poly(divinylbenzene) [poly(DVB)] column packing materials synthesized in the
first part of the study were tested in Reverse Phase Chromatography (RPC) under
different conditions. The investigation of column performances concluded that
columns packed with poly(VAc-co-DVB) (2500
plate/m) and poly(DVB) (1500
plate/m), showed high and stable theoretical plate number and high peak
resolution values compared to literature values. Protein recovery data in RPC
column, poly(VAc-co-DVB) (93-99%), were determined as similar as
quantitative values for all proteins. Reproducibility
tests of analytes of poly(VAc-co-DVB) column were assessed "run to run" and "day to day"
Relative Standard Deviation (RSD) values were determined under 2.8%.

Keywords

Monodisperse-Porous Particle Column Packed Material Reversed-Phase Chromatography (RPC) Protein Separation Protein Separation

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., 8 (1999) 875-889.
  • [2]. Ellingsten, T., Aune, O., Ugelstad, J., Hagen, S., Mono-sized Stationary Phases for Chromatography, Journal of Chromatography A, 535 (1990) 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., 32 (1994) 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., 73(24) (2001) 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., 20 (1997) 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, 66 (23) (1994) 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, 66 (13) (1994) 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, 383 (2005) 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, 570 (2006) 240-248 [10]. Ettre, L.S., Chromatography: the Separation Technique of the 20th Century. Chromatographia, 51 (1/2) (2000) 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, 800 (1-2) (2004) 3-6
  • [12]. Zhang, Y. W., Yao, S., Zeng, H., Song, H., Chiral Separation of Pharmaceuticals by High Performance Liquid Chromatography, Current Pharmaceutıcal Analysıs, 6 (2) (2010) 114-130
  • [13]. Saraçoğlu Kaya, B., Sağ, E., Design of Reversed Phase Chromatography (RPC) Column Packing Material for Protein Separation: Part 1- Synthesis and Characterization, Cumhuriyet Science Journal, 38 (4) (2017) 788-797
  • [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, 60 (2004) 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, 92 (2004) 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, 40 (2005) 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., 29 (2006) 936 – 944

Proteinleri Ayırmak İçin Ters Faz Kromatografisi Kolon Dolgu Materyali Tasarımı: Bölüm 2- Kromatografik Performans

Year 2019, , 213 - 220, 22.03.2019

Berna Saraçoğlu Kaya , Ebru Sağ

https://doi.org/10.17776/csj.364625

Abstract

İki bölümden oluşan çalışmanın ikinci
bölümünde çalışmanın ilk bölümünde sentezlenmiş olan eş boyutlu ve gözenekli
poli(vinil asetat-co-divinilbenzen) [poli(VAc-co-DVB)] ve poli(divinilbenzen)
[poli(DVB)] kolon dolgu materyalleri Ters
Faz Kromatografisi (Reversed Phase Chromatography, RPC) için farklı
koşullarda test edilmiştir. Sentez sonrası elde edilen partiküllerle doldurulan
kolonların kromatografik performansları incelediğinde, poli(VAc-co-DVB) (2500
tabaka/m) ve poli(DVB) (1500 tabaka/m) kolon
için literatürdeki değerlerden daha yüksek ve kararlı teorik tabaka sayısı
değerleri elde edilirken pik çözünürlük değerlerinin de kabul edilen değerlerin
üstünde olduğu görülmüştür. Poli(VAc-co-DVB) (%93-99) ulaşılan protein geri
kazanım değerleri ise bütün proteinler için kantitatife yakın değerler olarak
belirlenmiştir.  Üretilen kolonlar için
analitlerin, analizden analize ve günden güne tekrarlanabilirlik testleri
yapılmış, hem “analizden analize “ hem de “günden güne” Bağıl Standart Sapma
(BSS) değerleri % 2,8’in altında bulunmuştur.

Keywords

Eşboyutlu-Gözenekli Partikül Kolon Dolgu Materyali Ters Faz Kromatografisi (RPC) Protein Ayırma

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., 8 (1999) 875-889.
  • [2]. Ellingsten, T., Aune, O., Ugelstad, J., Hagen, S., Mono-sized Stationary Phases for Chromatography, Journal of Chromatography A, 535 (1990) 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., 32 (1994) 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., 73(24) (2001) 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., 20 (1997) 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, 66 (23) (1994) 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, 66 (13) (1994) 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, 383 (2005) 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, 570 (2006) 240-248 [10]. Ettre, L.S., Chromatography: the Separation Technique of the 20th Century. Chromatographia, 51 (1/2) (2000) 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, 800 (1-2) (2004) 3-6
  • [12]. Zhang, Y. W., Yao, S., Zeng, H., Song, H., Chiral Separation of Pharmaceuticals by High Performance Liquid Chromatography, Current Pharmaceutıcal Analysıs, 6 (2) (2010) 114-130
  • [13]. Saraçoğlu Kaya, B., Sağ, E., Design of Reversed Phase Chromatography (RPC) Column Packing Material for Protein Separation: Part 1- Synthesis and Characterization, Cumhuriyet Science Journal, 38 (4) (2017) 788-797
  • [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, 60 (2004) 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, 92 (2004) 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, 40 (2005) 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., 29 (2006) 936 – 944
There are 16 citations in total.

Details

Primary Language English
Journal Section Engineering Sciences
Authors

Berna Saraçoğlu Kaya 0000-0003-4706-5146

Ebru Sağ 0000-0001-5344-1705

Publication Date March 22, 2019
Submission Date December 11, 2017
Acceptance Date November 13, 2018
Published in Issue Year 2019

Cite

APA Saraçoğlu Kaya, B., & Sağ, E. (2019). Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance. Cumhuriyet Science Journal, 40(1), 213-220. https://doi.org/10.17776/csj.364625