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Advances in Ecdysteroid High Performance Liquid Chromatography

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Chromatography and Isolation of Insect Hormones and Pheromones

Part of the book series: Chromatographic Society Symposium Series ((CSSS))

Summary

High performance liquid chromatography (HPLC) is the most widely used method for the separation of ecdysteroids. Many efficient methods are presently available, which are adapted to the various classes of compounds (high, medium or low polarity ecdysteroids), for either analytical or preparative purposes.

This review includes a survey of classical chromatographic systems, including some recent developments of normal-phase HPLC (concerning especially the use of polar and non-polar bonded silica columns), a few general comments on rather sophisticated detection systems (diode-array detectors, radioactivity monitors) and some comments regarding the problem of sample preparation prior to HPLC analysis.

Several possible future developments of HPLC, including coupling with mass spectrometry, are discussed briefly.

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References

  1. R. Lafont and D. H. S. Horn, Phytoecdysteroids: structures and occurrence, in: “Ecdysone”, J. A. Koolman, ed., Georg Thieme-Verlag, Stuttgart, pp 39–64 (1989).

    Google Scholar 

  2. H. H. Rees, Zooecdysteroids: structures and occurrence, in “Ecdysone”, J. A. Koolman, ed., Georg Thieme-Verlag, Stuttgart, pp 28–38 (1989).

    Google Scholar 

  3. R. Lafont, P. Beydon, B. Mauchamp, G. Sommé-Martin, M. Andrianjafintrimo and P. Krien, Recent progress in ecdysteroid analytical methods, in: “Regulation of Insect Development and Behavior”, F. Sehnal, A. Zabza and B. Cymborowski, eds., Technical University of Wroclaw Press, pp 199–206 (1981).

    Google Scholar 

  4. J. C. Touchstone, Ecdysteroids, in: “CRC Handbook of Chromatography. Steroids”, CRC Press, Boca Raton, Florida, pp 119–135 (1986).

    Google Scholar 

  5. R Lafont, HPLC analysis of ecdysteroids in plants and animals, in: “Chromatography ‘87”, H. Kalasz and E. S. Ettre, eds., Akademia Kiado, Budapest, pp 1–15 (1988).

    Google Scholar 

  6. R Lafont, G. Sommé-Martin and J. C. Chambet, Separation of ecdysteroids using high-pressure liquid chromatography on microparticulate supports, J. Chromatog., 170:185–194(1979).

    Google Scholar 

  7. L. N. Dinan, P. L. Donnahey, H. H. Rees and T. W. Goodwin, High performance liquid chromatography of ecdysteroids and their 3-epi, 3-dehydro and 26-hydroxy derivatives, J. Chromatog., 205:139–145 (1981).

    Article  CAS  Google Scholar 

  8. B. Fournier and D. Radallah, Ecdysteroids in Carausius eggs during embryonic development, Archives Insect Biochem. & Physiol., 7:211–224 (1988).

    Article  CAS  Google Scholar 

  9. C. Blais and R. Lafont, Ecdysteroid metabolism by soluble enzymes from an insect: metabolic relationship between 3ß-hydroxy-, 3a-hydroxy- and 3-oxo-ecdysteroids, Hoppe-Seyler’s Zeitschrift Jïir Physiologische Chemie, 365:809–818 (1984).

    Article  CAS  Google Scholar 

  10. M. W. Raynor, J. P. Kithinji, K. D. Bartle, D. E. Games, I. C. Mylchreest, R Lafont, E. D. Morgan and I. D. Wilson, Packed column supercritical fluid chromatography and linked supercritical fluid chromatography — mass spectrometry for the analysis of phytoecdysteroids from Silène nutans and Silène otites, J. Chromatog., 467:292–298(1989).

    Google Scholar 

  11. P. Beydon, A. Fabre, J. Goichon and R Lafont, Use of a diode-array detector for HPLC analysis of ecdysteroids in biological extracts, Communication presented at the VIIIth Ecdysone Workshop, Marburg, FRG (1987).

    Google Scholar 

  12. R. E. Isaac, N. P. Milner and H. H. Rees, High-performance liquid chromatography of ecdysteroids and ecdysteroid 22-phosphates, J. Chromatog., 246:317–322 (1982).

    Article  CAS  Google Scholar 

  13. C. Hetru, B. Luu and J. A. Hoffmann, Ecdysone conjugates: isolation and identification, Methods in Enzymology, 11:411–419 (1985).

    Article  Google Scholar 

  14. R. E. Isaac, H. H. Rees and T. W. Goodwin, Isolation of 2-deoxy-20-hydroxyecdysone and 3-epi-2-deoxyecdysone from eggs of the desert locust, Schistocerca gregaria, J. Chem Soc., Chem. Communs., pp 418–420 (1981).

    Google Scholar 

  15. M. J. Thompson, G. F. Weirich, H. H. Rees, J. A. Svoboda, M. F. Feldlaufer and K. R. Wilzer, New ecdysteroid conjugate: isolation and identification of 26-hydroxyecdysone 26-phosphate from eggs of the tobacco hornworm, Manduca sexta (L.), Archives of Insect Biochem. & Physiol., 2:227–236 (1985).

    Article  CAS  Google Scholar 

  16. R. Lafont, G. Somme-Martin, B. Mauchamp, B. F. Maume and J. P. Delbecque, Analysis of ecdysteroids by high-performance liquid chromatography and coupled gas-liquid chromatography-mass spectrometry, in: “Progress in Ecdysone Research”, J. A. Hoffmann, ed., Elsevier Biomedical Press, Amsterdam, pp 45–68 (1980).

    Google Scholar 

  17. J. F. Modde, R. Lafont and J. A. Hoffmann, Ecdysone metabolism in Locusta migratoria larvae and adults, Internat J. Invertebrate Reprod. & Develop., 7:161–183 (1984).

    Article  CAS  Google Scholar 

  18. J. T. Warren, B. Steiner, A. Dorn, M. Pak and L. I. Gilbert, Metabolism of ecdysteroids during the embryogenesis of Manduca sexta, J. Liquid Chromatog., 9:1759–1782 (1986).

    Article  CAS  Google Scholar 

  19. P. Beydon, J. P. Girault and R. Lafont, Ecdysone metabolism in Pieris brassicae during the feeding last larval instar, Archives of Insect Biochem. & Physiol., 4:139–149 (1987).

    Article  CAS  Google Scholar 

  20. S. Scalia and E. D. Morgan, Simultaneous determination of free and conjugated ecdysteroids by liquid chromatography, J. Chromatog., 346:301–308 (1985).

    Article  CAS  Google Scholar 

  21. H. H. Rees and R. E. Isaac, Biosynthesis and metabolism of ecdysteroids and methods of isolation and identification of the free and conjugated ecdysteroids, Methods in Enzymology, 111:377–410(1985).

    Google Scholar 

  22. M. Hori, Automatic column chromatographic method for insect moulting steroids, Steroids, 14:33–45 (1969).

    Article  PubMed  CAS  Google Scholar 

  23. D. A. Schooley and K. Nakanishi, Applications of high-pressure liquid chromatography to the separation of insect moulting hormones, in: “Modern Methods of Steroid Analysis”, E. Heftmann, ed., Academic Press, New York, pp 37–54 (1973).

    Google Scholar 

  24. I. D. Wilson, C. R. Bielby and E. D. Morgan, Selective effects of mobile and stationary phases in reversed-phase high-performance liquid chromatography of ecdysteroids, J. Chromatog., 238:97–102 (1982).

    Article  CAS  Google Scholar 

  25. I. D. Wilson, C. R. Bielby and E. D. Morgan, Evaluation of some phytoecdysteroids as internal standards for the chromatographic analysis of ecdysone and 20- hydroxyecdysone from arthropods, J. Chromatog., 236:224–229 (1982).

    Article  CAS  Google Scholar 

  26. G. M. Holman and R. W. Meola, A high-performance liquid chromatography method for the purification and analysis of insect ecdysones: application to measurement of ecdysone titres during pupal-adult development of Heliothis zea, Insect Biochem., 8:275–278 (1978).

    Article  CAS  Google Scholar 

  27. I. Kubo and F. J. Hanke, Chemical methods for isolating and identifying phyto-chemicals biologically active in insects, in: “Insect-Plant Interactions — Springer Series in Experimental Entomology”, J. R. Miller and T. A. Miller, eds. Springer- Verlag, New York, pp 225–250 (1986).

    Chapter  Google Scholar 

  28. P. Nirde, G. Torpier, M. L. De Reggi and A. Capron, Ecdysone and 20-hydroxyecdysone: new hormones for the human parasite Schistosoma mansoni, J. Insect Physiol., 23:317–319 (1977).

    Article  Google Scholar 

  29. H. N. Nigg, M. J. Thompson, J. N. Kaplanis, J. A. Svoboda and W. E. Robbins, High- pressure liquid chromatography of the ecdysones — insect moulting hormones, Steroids, 23:507–516 (1974).

    Article  PubMed  CAS  Google Scholar 

  30. M. Valentin, W. E. Bollenbacher, L. I. Gilbert and H. Kroeger, Alterations in ecdysone content during the post-embryonic development of Chironomus thumni: corre lations with chromosomal puffing, Zeitschrift für Naturforschung, 33c:557–560 (1978).

    Article  CAS  Google Scholar 

  31. E. D. Morgan and C. F. Poole, The extraction and determination of ecdysones in Arthropods, Advances in Insect Physiol., 12:17–62 (1976).

    Article  CAS  Google Scholar 

  32. E. Ohnishi, T. Mizuno, N. Ikekawa, N. Awata and S. Sakurai, Occurrence of oc-ecdysone in the developing embryos of the silkworm, Bombyx mori, J. Insect Physiol., 23:317–319 (1977).

    Article  CAS  Google Scholar 

  33. B. Stay, L. S. Ostedgaard, S. S. Tobe, A. Strambi and E. Spaziani, Ovarian and haemolymph titres of ecdysteroid during the gonadotrophic cycle in Diploptera punctata, J. Insect Physiol., 30:643–651 (1984).

    Article  CAS  Google Scholar 

  34. A. Moribayashi, H. Kurahashi and T. Ohtaki, Comparative studies on ecdysone metabolism between mature larvae and pharate pupae in the fleshfly, Sarcophaga peregrina, Archives Insect Biochem. & Physiol., 2:237–250 (1985).

    Article  CAS  Google Scholar 

  35. K. H. Hoffmann, D. Bulenda, E. Thiiy and E. Schmid, Apolar ecdysteroid esters in adult female crickets, GryRus bimaculatus, Life Sciences, 37:185–192 (1985).

    Article  PubMed  CAS  Google Scholar 

  36. T. Crosby, R. P. Evershed, D. Lewis, K. P. Wigglesworth and H. H. Rees, Identification of ecdysone 22-long-chain fatty acyl esters in newly laid eggs of the cattle tick Boophilus microplus, Biochem. J., 240:131–138 (1986).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  37. P. A. Diehl, J. L. Connat, J. P. Girault and R. Lafont, A new class of apolar ecdysteroid conjugates: esters of 20-hydroxyecdysone with long-chain fatty acids in ticks, Internat J. Invertebrate Reprod. & Develop., 8:1–13 (1985).

    Article  CAS  Google Scholar 

  38. I. Kubo, S. Komatsu, I. Asaka and G. De Boer, Isolation and identification of apolar metabolites of ingested 20-hydroxyecdysone in frass of Heliothis virescens larvae, J. Chem. Ecol., 13:785–794 (1987).

    Article  PubMed  CAS  Google Scholar 

  39. J. P. Girault, C. Blais, P. Beydon, C. Rolando and R. Lafont, Synthesis and NMR study of 3-dehydroecdysteroids, Archives Insect Biochem. & Physiol., 10:199–213(1989).

    Google Scholar 

  40. S. Terabe, Electrokinetic chromatography: an interface between electrophoresis and chromatography, Trends Anal. Chem., 8:129–134 (1989).

    Article  CAS  Google Scholar 

  41. I. D. Wilson, E. D. Morgan, P. Robinson, R. Lafont and C. Biais, A comparison of radio-thin-layer and radio-high-performance liquid chromatography for ecdysteroid metabolism studies, J. Insect Physiol., 34:707–711 (1988).

    Article  CAS  Google Scholar 

  42. C. F. Poole, S. Singhawangcha, A. Zlatkis and E. D. Morgan, Polynuclear aromatic boronic acids as selective fluorescent reagents for HPTLC and HPLC, J. High Res. Chrom. & Chrom. Commun., 1:96–97 (1978).

    Article  CAS  Google Scholar 

  43. I. Kubo and S. Komatsu, Micro analysis of prostaglandins and ecdysteroids in insects by high-performance liquid chromatography and fluorescence labeling, J. Chromatog., 362:61–70 (1986).

    Article  CAS  Google Scholar 

  44. T. Takemoto, S. Ogawa, N. Nishimoto, K. Y. Yen, K. Abe, T. Sato, K. Ogawa and M. Takahashi, The Isolation of ecdysterone from the radix of Achyranthes obtusifolia Lam, Yakugaku Zasshi, 87:1521–1524 (1967).

    Article  PubMed  CAS  Google Scholar 

  45. M. W. Gilgan and M. E. Zinck, Estimation of ecdysterone from sulphuric acid-induced fluorescence, Steroids, 20:95–104 (1972).

    Article  PubMed  CAS  Google Scholar 

  46. R. T. Mayer and J. A. Svoboda, Thin-layer chromatographic in situ analysis of insect ecdysones via fluorescence quenching, Steroids, 31:139–150 (1978).

    Article  PubMed  CAS  Google Scholar 

  47. J. Koolman, Analysis of ecdysteroids by fluorometry, Insect Biochem., 10:381–386 (1980).

    Article  CAS  Google Scholar 

  48. R. Lafont and P. Beydon, Methods for ecdysteroid analysis, in: “Recent Advances in Comparative Arthropod Morphology, Physiology and Development”, A. P. Gupta, ed., Rutgers University Press, Vol. 1, pp 455–512 (in press) (1989).

    Google Scholar 

  49. E. D. Morgan and I. D. Wilson, Methods for separation and physico-chemical quan tification of ecdysteroids, in: “Ecdysone”, J. A. Koolman, ed., Georg Thieme-Verlag, Stuttgart, pp 114–130 (1989).

    Google Scholar 

  50. D.A. Schooley, G. Weiss and K. Nakanishi, A simple and general extraction procedure for phytoecdysones based on reversed-phase adsorption chromatography, Steroids, 19:377–383(1973).

    Article  Google Scholar 

  51. I. D. Wilson, C. R. Bielby, E. D. Morgan and A. E. M. McLean, Comparison of high- performance liquid chromatography and gas-chromatography for the analysis of ecdysteroids, J. Chromatog., 194:343–352 (1980).

    Article  CAS  Google Scholar 

  52. R. D. Watson and E. Spaziani, Rapid isolation of ecdysteroids from crustacean tissues nd culture media using Sep-pak C18 cartridges, J. Liquid Chromatog., 5:525–535 (1982).

    Article  CAS  Google Scholar 

  53. G. D. Pimprikar, M. J. Coign, H. Sakurai and J. R. Heitz, High-performance liquid chromatographic determination of ecdysteroid titers in the house fly, J. Chromatog., 317:413–419 (1984).

    Article  CAS  Google Scholar 

  54. R. Lafont, J. L. Pennetier, A. Andrianjafintrimo, J. Claret, J. F. Modde and C. Biais, Sample processing for high-performance liquid chromatography of ecdysteroids, J. Chromatog., 236:137–149 (1982).

    Article  CAS  Google Scholar 

  55. I. Kubo, A. Matsumoto and S. Asano, Efficient isolation of ecdysteroids from the silk worm, Bombyx mori by droplet counter-current chromatography, Insect Biochem., 15:45–47 (1985).

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Department de Biologie, Ecole Normale Supérieure, CNRS-URA 686, Université Paris, F-75230, Paris Cedex 05, France

    R. Lafont

  2. Department of Chemistry, University of Keele, Keele, Staffordshire, ST5 5BG, UK

    I. D. Wilson

Authors
  1. R. Lafont
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  2. I. D. Wilson
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Editor information

Editors and Affiliations

  1. University of Reading, Reading, UK

    A. R. McCaffery

  2. ICI Pharmaceuticals, Macclesfield, Cheshire, UK

    I. D. Wilson

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© 1990 Plenum Press, New York

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Lafont, R., Wilson, I.D. (1990). Advances in Ecdysteroid High Performance Liquid Chromatography. In: McCaffery, A.R., Wilson, I.D. (eds) Chromatography and Isolation of Insect Hormones and Pheromones. Chromatographic Society Symposium Series. Springer, New York, NY. https://doi.org/10.1007/978-1-4684-8062-7_8

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  • DOI: https://doi.org/10.1007/978-1-4684-8062-7_8

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4684-8064-1

  • Online ISBN: 978-1-4684-8062-7

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