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Highly hydrophilic and nonionic poly(2-vinyloxazoline)-grafted silica: a novel organic phase for high-selectivity hydrophilic interaction chromatography

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Abstract

A new hydrophilic and nonionic poly(2-vinyloxazoline)-grafted silica (Sil-VOX n ) phase was synthesized and applied for the separation of nucleosides and nucleobases in hydrophilic interaction chromatography (HILIC). Polymerization and immobilization onto silica were confirmed by using characterization techniques including 1H NMR spectroscopy, elemental analysis, and diffuse reflectance infrared Fourier transform spectroscopy. The hydrophilicity or wettability of Sil-VOX n was observed by measuring the contact angle (59.9°). The chromatographic results were compared with those obtained with a conventional HILIC silica column. The Sil-VOX n phase showed much better separation of polar test analytes than the silica column, and the elution order was different. Differences in selectivity between these two columns indicate that the stationary phase cannot function merely as an inert support for a water layer into which the solutes are partitioned from the bulk mobile phase. To elucidate the interaction mechanism, the separation of dihydroxybenzene isomers was performed on both columns in normal-phase liquid chromatography. Sil-VOX n was very sensitive to the dipole moments of the positional isomers of polycyclic aromatic compounds in normal-phase liquid chromatography. The interaction mechanism for Sil-VOX n in HILIC separation is also described.

Separation of nucleosides and nucleobases with Sil-VOXn (bottom) and a commercial silica column (top). Mobile phase of acetonitrile and 20 mM ammonium acetate (9:1, v/v). Flow rate 1 ml min-1, column temperature 25 °C. The analytes were as follows 5-iodouracil (1), thymine (2), uracil (3), 4,6-diaminopyrimidine (4), uridine (5), adenosine 2 (6), cytosine (7), cytidine (8), and guanosine (9)

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References

  1. Sander LC, Wise SA (1984) Synthesis and characterization of polymeric C18 stationary phases for liquid chromatography. Anal Chem 56:504–510

    Article  CAS  Google Scholar 

  2. Sentell KB, Dorsey JG (1989) Retention mechanisms in reversed-phase liquid chromatography. Stationary-phase bonding density and partitioning. Anal Chem 61:930–934

    Article  CAS  Google Scholar 

  3. KimataK HT, Morichi K, Hosoya K, Araki T, Tanaka N (1995) High-capacity stationary phases containing heavy atoms for HPLC separation of fullerenes. Anal Chem 67:2556–2561

    Article  Google Scholar 

  4. Jinno K, Tanabe K, Saito Y, Nagashima H (1997) Separation of polycyclic aromatic hydrocarbons with various C60 fullerene bonded silica phases in microcolumn liquid chromatography. Analyst 122:787–791

  5. Mallik AK, Qiu H, Takafuji M, Ihara H (2012) Selectivity enhancement for the separation of tocopherols and steroids by integration of highly ordered weak interaction sites along the polymer main chain. Anal Bioanal Chem 404:229–238

    Article  CAS  Google Scholar 

  6. Qiu H, Mallik AK, Takafuji M, Liu X, Jiang S, Ihara H (2012) A new imidazolium-embedded C18 stationary phase with enhanced performance in reversed-phase liquid chromatography. Chim Acta 738:95–101

    Article  CAS  Google Scholar 

  7. Jiang Z, Smith NW, Ferguson PD, Taylor MR (2009) Novel highly hydrophilic zwitterionic monolithic column for hydrophilic interaction chromatography. J Sep Sci 32:2544–2555

    Article  CAS  Google Scholar 

  8. Wang XC, Lu HX, Lin XC, Xie ZH (2008) Electrochromatographic characterization of methacrylate-based monolith with mixed mode of hydrophilic and weak electrostatic interactions by pressurized capillary electrochromatography. J Chromatogr A 1190:365–371

    Article  CAS  Google Scholar 

  9. McCalley DV (2010) The challenges of the analysis of basic compounds by high performance liquid chromatography: Some possible approaches for improved separations. J Chromatogr A 1217:858–880

    Article  CAS  Google Scholar 

  10. Alpert AJ (1990) Hydrophilic-interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. J Chromatogr 499:177–196

    Article  CAS  Google Scholar 

  11. Jin HL, Stalcup AM, Armstrong DW (1988) Separation of cyclodextrins using cyclodextrin bonded phases. J Liq Chromatogr 11:3295–3304

  12. Churms SC (1995) In: El Rassi Z (ed) Carbohydrate analysis. Journal of chromatography library, vol 58. Elsevier, New York, pp 103–146

  13. Armstrong DW, Jin HL (1989) Evaluation of the liquid chromatographic separation of monosaccharides, disaccharides, trisaccharides, tetrasaccharides, deoxysaccharides and sugar alcohols with stable cyclodextrin bonded phase columns. J Chromatogr A 462:219–232

    Article  CAS  Google Scholar 

  14. Yoshida T (2004) Peptide separation by hydrophilic-interaction chromatography: a review. J Biochem Biophys Methods 60:265–280

    Article  CAS  Google Scholar 

  15. Lam MP, Siu S, Lau E, Mao X, Sun H, Chiu PC, Yeung WS, Cox DM, Chu IK (2010) Online coupling of reverse-phase and hydrophilic interaction liquid chromatography for protein and glycoprotein characterization. Anal Bioanal Chem 398:791–804

    Article  CAS  Google Scholar 

  16. Strege MA (1998) Hydrophilic interaction chromatography-electrospray mass spectrometry analysis of polar compounds for natural product drug discovery. Anal Chem 70:2439–2445

    Article  CAS  Google Scholar 

  17. StregeMA SS, Lawrence SM (2000) Mixed-mode anion-cation exchange/hydrophilic interaction liquid chromatography-electrospray mass spectrometry as an alternative to reversed phase for small molecule drug discovery. Anal Chem 72:4629–4633

    Article  Google Scholar 

  18. Guo Y, Gaiki S (2005) Retention behavior of small polar compounds on polar stationary phases in hydrophilic interaction chromatography. J Chromatogr A 1074:71–80

    Article  CAS  Google Scholar 

  19. Kaliszan R (2007) QSRR: quantitative structure-(chromatographic) retention relationships. Chem Rev 107:3212–3246

    Article  CAS  Google Scholar 

  20. Buszewski B, Noga S (2012) Hydrophilic interaction liquid chromatography (HILIC)–a powerful separation technique. Anal Bioanal Chem 402:231–247

    Article  CAS  Google Scholar 

  21. Hemström P, Irgum K (2006) Hydrophilic interaction chromatography. J Sep Sci 29:1784–1821

    Article  Google Scholar 

  22. Dinh NP, Jonsson T, Irgum K (2011) Probing the interaction mode in hydrophilic interaction chromatography. J Chromatogr A 1218:5880–5891

    Article  CAS  Google Scholar 

  23. Qiu H, Wanigasekara E, Zhang Y, Tran T, Armstrong DW (2011) Development and evaluation of new zwitterionic Hydrophilic interaction liquid chromatography stationary phases based on 3-P, P-diphenylphosphonium-propylsulfonate. J Chromatogr A 1218:8075–8082

    Article  CAS  Google Scholar 

  24. Weng N, Shou W, Chen YL, Jiang X (2001) Novel liquid chromatographic-tandem mass spectrometric methods using silica columns and aqueous-organic mobile phases for quantitative analysis of polar ionic analytes in biological fluids. J Chromatogr B 754:387–399

    Article  Google Scholar 

  25. Weng N, Shou WZ, Addison T, Maleki S, Jiang X (2002) Liquid chromatography/tandem mass spectrometric bioanalysis using normal-phase columns with aqueous/organic mobile phases - a novel approach of eliminating evaporation and reconstitution steps in 96-well SPE. Rapid Commun Mass Spectrom 16:1965–1975

    Article  Google Scholar 

  26. Naidong W, Eerkes A (2004) Development and validation of a hydrophilic interaction liquid chromatography-tandem mass spectrometric method for the analysis of paroxetine in human plasma. Biomed Chromatogr 18:28–36

    Article  CAS  Google Scholar 

  27. Guo Y, Huang AJ (2003) A HILIC method for the analysis of tromethamine as the counter ion in an investigational pharmaceutical salt. Pharm Biomed Anal 31:1191–1201

    Article  CAS  Google Scholar 

  28. Yoshida T (1997) Peptide separation in normal phase liquid chromatography. Anal Chem 69:3038–3043

    Article  CAS  Google Scholar 

  29. Karatapanis AE, Fiamegos YC, Stalikas CD (2010) Study of the behavior of water-soluble vitamins in HILIC on a diol column. Chromatographia 71:751–759

    Article  CAS  Google Scholar 

  30. Berthod A, Chang SS, Kullman JP, Armstrong DW (1998) Practice and mechanism of HPLC oligosaccharide separation with a cyclodextrin bonded phase. Talanta 47:1001–1012

    Article  CAS  Google Scholar 

  31. Jiang W, Irgum K (1999) Covalently bonded polymeric zwitterionic stationary phase for simultaneous separation of inorganic cations and anions. Anal Chem 71:333–344

  32. Moni L, Ciogli A, D’Acquarica I, Dondoni A, Gasparrini F, Marra A (2010) Synthesis of sugar-based silica gels by copper-catalysed azide-alkyne cycloaddition via a single-step azido-activated silica intermediate and the use of the gels in hydrophilic interaction chromatography. Chem Eur J 57:12–22

    Google Scholar 

  33. Sedlacek O, Monnery BD, Filippov SK, Hoogenboom R, Hruby M (2012) Poly(2-oxazoline)s–are they more advantageous for biomedical applications than other polymers? Macromol Rapid Commun 33:1648–1662

    Article  CAS  Google Scholar 

  34. Makino A, Kobayashi S (2010) Chemistry of 2-oxazolines: A crossing of cationic ring-opening polymerization and enzymatic ring-opening polyaddition. J Polym Sci Part A 48:1251–1270

    Article  CAS  Google Scholar 

  35. Mallik AK, Takafuji M, Ihara H (2009) Molecular-shape selectivity tuned by donor–acceptor type copolymers as organic phase in reversed-phase high-performance liquid chromatography. J Chromatogr A 1216:7433–7439

    Article  CAS  Google Scholar 

  36. Hurwitz G, Guillen GR, Hoek E (2010) Probing polyamide membrane surface charge, zeta potential, wettability, and hydrophilicity with contact angle measurements. J Membr Sci 349:349–357

    Article  CAS  Google Scholar 

  37. Shundo A, Hori K, Ikeda T, Kimizuka N, Tanaka K (2013) Design of a dynamic polymer interface for chiral discrimination. J Am Chem Soc 135:10282–10285

    Article  CAS  Google Scholar 

  38. Gama MR, da Costa Silva RG, Collins CH, Bottoli CB (2012) Hydrophilic interaction chromatography. Trends Anal Chem 37:48–60

    Article  CAS  Google Scholar 

  39. McCalley DV (2010) Study of the selectivity, retention mechanisms and performance of alternative silica-based stationary phases for separation of ionised solutes in hydrophilic interaction chromatography. J Chromatogr A 1217:3408–3417

    Article  CAS  Google Scholar 

  40. Lü Z, Zhang P, Jia L (2010) Preparation of chitosan functionalized monolithic silica column for hydrophilic interaction liquid chromatography. J Chromatogr A 1217:4958–4964

    Article  Google Scholar 

  41. Kiselev AV, Aratskova AA, Gvozdovltch TN, Yashin YI (1980) Retention behaviour of o-, m- and p-isomers of benzene derivatives on a silica gel hydroxylated surface in liquid chromatography. J Chromatogr 195:205–210

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Grant-in-Aid for Regional Innovation Strategy Support Program and in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan. The authors thank Kojin Film and Chemicals for proving the 2-vinyloxazoline monomer as a gift.

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Correspondence to Hirotaka Ihara.

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Mallik, A.K., Cheah, W.K., Shingo, K. et al. Highly hydrophilic and nonionic poly(2-vinyloxazoline)-grafted silica: a novel organic phase for high-selectivity hydrophilic interaction chromatography. Anal Bioanal Chem 406, 4585–4593 (2014). https://doi.org/10.1007/s00216-014-7868-7

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  • DOI: https://doi.org/10.1007/s00216-014-7868-7

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