Year 2019, Volume 40, Issue 1, Pages 213 - 220 2019-03-22

Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance
Proteinleri Ayırmak İçin Ters Faz Kromatografisi Kolon Dolgu Materyali Tasarımı: Bölüm 2- Kromatografik Performans

Berna SARAÇOĞLU KAYA [1] , Ebru SAĞ [2]

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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%.

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.

  • [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
Primary Language en
Subjects Basic Sciences
Journal Section Engineering Sciences
Authors

Orcid: 0000-0003-4706-5146
Author: Berna SARAÇOĞLU KAYA (Primary Author)
Country: Turkey


Orcid: 0000-0001-5344-1705
Author: Ebru SAĞ
Country: Turkey


Dates

Publication Date: March 22, 2019

Bibtex @research article { csj364625, journal = {Cumhuriyet Science Journal}, issn = {2587-2680}, eissn = {2587-246X}, address = {Cumhuriyet University}, year = {2019}, volume = {40}, pages = {213 - 220}, doi = {10.17776/csj.364625}, title = {Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance}, key = {cite}, author = {SARAÇOĞLU KAYA, Berna and SAĞ, Ebru} }
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. DOI: 10.17776/csj.364625
MLA SARAÇOĞLU KAYA, B , SAĞ, E . "Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance". Cumhuriyet Science Journal 40 (2019): 213-220 <http://dergipark.org.tr/csj/issue/43798/364625>
Chicago SARAÇOĞLU KAYA, B , SAĞ, E . "Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance". Cumhuriyet Science Journal 40 (2019): 213-220
RIS TY - JOUR T1 - Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance AU - Berna SARAÇOĞLU KAYA , Ebru SAĞ Y1 - 2019 PY - 2019 N1 - doi: 10.17776/csj.364625 DO - 10.17776/csj.364625 T2 - Cumhuriyet Science Journal JF - Journal JO - JOR SP - 213 EP - 220 VL - 40 IS - 1 SN - 2587-2680-2587-246X M3 - doi: 10.17776/csj.364625 UR - https://doi.org/10.17776/csj.364625 Y2 - 2018 ER -
EndNote %0 Cumhuriyet Science Journal Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance %A Berna SARAÇOĞLU KAYA , Ebru SAĞ %T Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance %D 2019 %J Cumhuriyet Science Journal %P 2587-2680-2587-246X %V 40 %N 1 %R doi: 10.17776/csj.364625 %U 10.17776/csj.364625
ISNAD SARAÇOĞLU KAYA, Berna , SAĞ, Ebru . "Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance". Cumhuriyet Science Journal 40 / 1 (March 2019): 213-220. https://doi.org/10.17776/csj.364625
AMA SARAÇOĞLU KAYA B , SAĞ E . Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance. CSJ. 2019; 40(1): 213-220.
Vancouver SARAÇOĞLU KAYA B , SAĞ E . Design of Reversed-Phase Chromatography Column Packing Material for Protein Separation: Part 2- Chromatographic Performance. Cumhuriyet Science Journal. 2019; 40(1): 220-213.