, Volume 67, Issue 9–10, pp 767–772 | Cite as

Dynamic On-Column Eluent Modification: A Novel Strategy for Peak Resolution Enhancement. Application to the Preparative Separation of Ecdysteroid Isomers

Full Short Communication


A novel LC method was applied to enhance the peak resolution of two ecdysteroids: 20-hydroxyecdysone and its 3-epimer. An isocratic solvent system of methanol–water (6:4, v/v) was used on a C18 column, and 100 μL of water was injected during the development in such a manner that the eluted solvent peak appeared exactly between the two overlapping peaks. This resulted in the increased retardation of the later-eluted peak, and in a good separation of the two compounds within 4.5 min.


Column liquid chromatography Preparative LC Ecdysteroid 20-Hydroxyecdysone and 3-epi-20-hydroxyecdysone 


  1. 1.
    Dinan L, Savchenko T, Whiting P (2001) Cell Mol Life Sci 58:1121–1132CrossRefGoogle Scholar
  2. 2.
    Lafont R, Dinan L (2003) J Ins Sci 3(7):1–30Google Scholar
  3. 3.
    Slama K, Lafont R (1995) Eur J Entomol 92:355–377Google Scholar
  4. 4.
    Bathori M, Pongracz Z (2005) Curr Med Chem 12:153–172Google Scholar
  5. 5.
    Dinan L (2001) Phytochem 57:325–339CrossRefGoogle Scholar
  6. 6.
    Lafont R, Horn DHS (1989) Phytoecdysteroids: structures and occurrence. In: Koolman J (ed) Ecdysone—from chemistry to mode of action. Georg Thieme Verlag, Stuttgart, New York, pp 39–64Google Scholar
  7. 7.
    Meng Y, Whiting P, Zibareva L, Bertho G, Girault JP, Lafont R, Dinan L (2001) J Chromatogr A 935:309–319CrossRefGoogle Scholar
  8. 8.
    Yagasinghe L, Jayasooriya K, Oyama K, Fujimoto Y (2002) Steroids 67:555–558CrossRefGoogle Scholar
  9. 9.
    Lafont R, Blais C, Harmatha J, Wilson ID (2000) Ecdysteroids; chromatography. In: Wilson ID, Adlard ER, Cooke M, Poole CF (eds) Encyclopedia of separation science. Academic Press, London, pp 2631–2643Google Scholar
  10. 10.
    Dinan L, Harmatha J, Lafont R (2001) J Chromatogr A 935:105–123CrossRefGoogle Scholar
  11. 11.
    Lafont R, Beydon P (1990) Methods for ecdysteroid analysis, Chap. 13. In: Gupta AP (ed) Morphogenetic hormones of arthropods. Discoveries, syntheses, metabolism, evolution, modes of action and techniques. Rutgers University Press, New Brunswick, London, pp 455–512Google Scholar
  12. 12.
    Evershed RP, Kabbouh M, Prescott MC, Maggs JL, Rees HH (1990) Current status and recent advances in the chromatography and mass spectrometry of ecdysteroids. In: McCaffery AR, Wilson ID (eds) Chromatography and isolation of insect hormones and pheromones. Plenum Press, New York, London, pp 103–115Google Scholar
  13. 13.
    Lafont R, Harmatha J, Dinan L, Wilson ID (eds) Ecdybase. The Ecdysone Handbook, 3rd edn. Accessed 2002
  14. 14.
    Lafont R, Kaouadji N, Morgan ED, Wilson ID (1994) J Chromatogr 658:55–67CrossRefGoogle Scholar
  15. 15.
    Lafont R, Pennetier JL, Andrianjafintrimo M, Claret J, Modde JF, Blais C (1982) J Chromatogr 236:137–149CrossRefGoogle Scholar
  16. 16.
    Kayser H, Eilinger P (1999) Arch Insect Biochem Physiol 41:162–170CrossRefGoogle Scholar
  17. 17.
    Li J, Qi H, Qi LW, Yi L, Li P (2007) Anal Chim Acta 596:264–272CrossRefGoogle Scholar
  18. 18.
    Hunyadi A, Gergely A, Simon A, Tóth G, Veress G, Báthori M (2007) J Chrom Sci 45:76–86Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlag/GWV Fachverlage GmbH 2008

Authors and Affiliations

  1. 1.Department of PharmacognosyUniversity of SzegedSzegedHungary
  2. 2.Department of Pharmacology and PharmacotherapySemmelweis UniversityBudapestHungary

Personalised recommendations