, Volume 66, Issue 3–4, pp 223–229 | Cite as

Investigating the Effect of Chromatographic Conditions on Retention of Organic Acids in Hydrophilic Interaction Chromatography Using a Design of Experiment



Small organic acids have shown significant retention on various stationary phases, such as amide, amino, aspartamide, silica and sulfobetaine phase commonly used in hydrophilic interaction chromatography (HILIC). This study investigated the effect of chromatographic conditions on the retention behavior of organic acids in HILIC using the tool of design of experiment (DOE). The results of the DOE study indicated that both the content of organic solvent (i.e., acetonitrile) and salt concentration in the mobile phase had significant effects on the retention of organic acids. Higher content of organic solvent in the mobile phase led to a significant increase in retention on all types of stationary phases. Increasing salt concentration also resulted in a moderate increase in retention; however, the effect of salt concentration varied with the type of stationary phase. The study also revealed that column temperature had less impact on retention than organic solvent content and salt concentration in HILIC.


Hydrophilic interaction chromatography Design of experiment Organic acid 



The authors would like to acknowledge the Global ChemPharm management for supporting the co-op program, in which Sriram Srinivasan participated.


  1. 1.
    Hemstrom P, Irgum K (2006) J Sep Sci 29:1784–1821CrossRefGoogle Scholar
  2. 2.
    Naidong W (2003) J Chromatogr B 796:209–224CrossRefGoogle Scholar
  3. 3.
    Bajad SU, Lu W, Kimball EH, Yuan J, Peterson C, Rabinowitz JD (2006) J Chromatogr A 1125:76–88CrossRefGoogle Scholar
  4. 4.
    Xue YJ, Liu J, Unger S (2006) J Pharm Biomed Anal 41:979–988CrossRefGoogle Scholar
  5. 5.
    Gavin PF, Olsen BA, Wirth DD, Lorenz KT (2006) J Pharm Biomed Anal 41:1251–1259CrossRefGoogle Scholar
  6. 6.
    Risley DS, Yang WQ, Peterson JA (2006) J Sep Sci 29:256–264CrossRefGoogle Scholar
  7. 7.
    Wang X, Li W, Rasmussen HT (2005) J Chromatogr A 1083:58–62CrossRefGoogle Scholar
  8. 8.
    Ciminiello P, Dell’Aversano C, Fattorusso E, Forino M, Martino M, Magno S, Santelia F, Tsoukatou M (2006) Toxicon 47:174–181CrossRefGoogle Scholar
  9. 9.
    Dell’Aversano C, Hess P, Quilliam MA (2005) J Chromatogr A 1081:190–201CrossRefGoogle Scholar
  10. 10.
    Schlichtherle-Cerny H, Affloter M, Cerny C (2004) Food Sci Tech 131:363–378Google Scholar
  11. 11.
    Schlichtherle-Cerny H, Affloter M, Cerny C (2003) Anal Chem 75:2349–2354CrossRefGoogle Scholar
  12. 12.
    Li AC, Chen YL, Junga H, Shou WZ, Jiang X, Naidong W (2003) Chromatographia 58:723–731Google Scholar
  13. 13.
    Hiseh Y, Chen J (2005) Rapid Comm Mass Spec 19:3031–3036CrossRefGoogle Scholar
  14. 14.
    Appelblad P, Abrahamsson P (2005) In: The application book March 2005 LC-GC Europe, pp 47–48Google Scholar
  15. 15.
    Delinsky AD, Delinsky DC, Muralidhara S, Fisher JW, Bruckner JV, Bartlett MG (2005) Rapid Comm Mass Spec 19:1075–1083CrossRefGoogle Scholar
  16. 16.
    Guo Y, Gaiki S (2005) J Chromatogr A 1074:71–80CrossRefGoogle Scholar
  17. 17.
    Grumbach ES, Wagrowski-Diehl DM, Mazzeo JR, Alden B, Iraneta PC (2004) LC-GC North America 7:92–98Google Scholar
  18. 18.
    Mathews P (2005) In: Design of Experiments with MiniTAB, ASQ Quality Press, Milwaukee, WisconsinGoogle Scholar
  19. 19.
    Ye C, Liu J, Ren F, Okafo N (2000) J Pharm Biomed Anal 23:581–589CrossRefGoogle Scholar
  20. 20.
    Ferreiros N, Iriate G, Alonso RM, Jimenez RM (2006) Talanta 69:747–756CrossRefGoogle Scholar
  21. 21.
    Dejaegher B, Jimidar M, De Smet M, Cockaerts P, Smeyers-Verbeke J, Vander Heyden Y (2006) J Pharm Biomed Anal 42:155–170CrossRefGoogle Scholar
  22. 22.
    Belfast M, Lu R, Capen R, Zhong J, Nguyen M, Gimenez J, Sitrin R, Mancinelli R (2006) J Chromatogr B 832:208–215CrossRefGoogle Scholar
  23. 23.
    Alpert AJ (1999) Size exclusion chromatography of small solutes. In: Wu CS (eds) Column handbook for size exclusion chromatography. Academic, San Diego, pp 249–266Google Scholar
  24. 24.
    Alpert AJ (1990) J Chromatogr 499:177–196CrossRefGoogle Scholar
  25. 25.
    Naidong W, Chen Y-L, Shou WZ, Jiang X (2001) J Biomed Anal 26:753–767CrossRefGoogle Scholar
  26. 26.
    Scott RPW The Thermodynamics of Chromatography

Copyright information

© Friedr. Vieweg & Sohn Verlag/GWV Fachverlage GmbH 2007

Authors and Affiliations

  1. 1.Analytical Development/Global ChemPharm DevelopmentJohnson & Johnson Pharmaceutical Research and Development, L.L.CRaritanUSA
  2. 2.Analytical R&D, MicroDose TechnologiesMonmouth JunctionUSA

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