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Mobile phase compositions for ceramide III by normal phase high performance liquid chromatography

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Abstract

Ceramide III was prepared by the cultivation ofSaccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by Normal Phase High Performance Liquid Chromatography (NP-HPLC) using Evaporative Light Scattering Detector (ELSD). We experimentally determined the mobile phase composition to separate ceramide III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol,n-hexane/Isoprophyl Alcohol (IPA) andn-hexane/n-butanol and one ternary mobile phase ofn-hexane/IPA/methanol were demonstrated. For the binary mobile phase ofn-hexane/ethanol, the first mobile phase composition, 95/5 (v/v), was step-increased to 72/23 (v/v) at 3 min. In the binary mobile phase, the retention time of ceramide III was 7.87 min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition ofn-hexane/IPA/methanol, 85/7/8 (v/v/v), was step-increased to 75/10/15 (v/v/v) at 3 min. However, in the ternary mobile phase, the more peak area of ceramide III was observed.

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References

  1. [1]

    Bouwstra, J. A., K. Cheng, G. S. Gooris, A. Weerheim. and M. Ponec (1996) The role of ceramides 1 and 2 in the stratum corneum lipid organisation.Biochim. Biophys. Acta 1300: 177–186.

  2. [2]

    Raith, K. and H. H. N. Reinhard (2000) Liquid chromatography-electrospray mass spectrometry and tandem mass spectrometry of ceramides.Anal. Chim. Acta 403: 295–303.

  3. [3]

    Rupeic, J. and V. Maric (1998) Isolation and chemical composition of the ceramide of theCandida lipolytica yeast.Chem. Phys. Lipids 91: 153–161.

  4. [4]

    Aida, E. C. and S. F. Anthony (2000) Current methods for the identification and quantitation of ceramides: An overview.Lipids 35: 937–945.

  5. [5]

    Rupeic, J., M. Mesaric, and V. Maric (1998) The influence of carbon source on the level and composition of ceramides of theCandida lipolytica yeast.Appl. Microbiol. Biotechnol. 50: 583–588.

  6. [6]

    Gildenast, T. and J. Lasch (1997) Isolation of ceramide fractions from human stratum corneum lipid extracts by high-performance liquid chromatography.Biochim. Biophys. Acta 1346: 69–74.

  7. [7]

    Gaudin, K., E. Lesellier, P. Chaminade, D. Ferrier, A. Baillet, and A. Tchapla (2000) Retention behaviour of ceramides in sub-critical fluid chromatography in comparison with non-aqueous reversed-phase liquid chromatography.J. Chromatogr. A 883: 211–222.

  8. [8]

    Molander, P., A. Holm, E. Lundanes, and T. Greibrokk (2000) Separation of ceramides by sub-ambient temperature-assisted large volume injection in temperature-programmed packed capillary liquid chromatography.J. High Resol. Chromatogr. 23: 653–655.

  9. [9]

    Wells, G., R. Dickson, and R. Lester (1998) Heat-induced elevation of ceramide inSaccharomyces cerevisiae via de novo synthesis.J. Biol. Chem. 273: 7235–7243.

  10. [10]

    Thomas, J. M., E. C. Aida, R.B. Phyllis, and S. F. Anthony (1999) The separation and direction detection of ceramide and sphingoid bases by normal-phase high performance liquid chromatography and evaporative light-scattering detection.Analy. Biochem. 276: 242–250.

  11. [11]

    Raith, K., J. Darius, and R. H. H. Neubert (2000) Ceramide analysis utilizing gas chromatography-mass spectrometry.J. Chromatogr. A 876: 229–233.

  12. [12]

    Long, A. S., P. W. Wertz, J. S. Strauss, and D. T. Downing (1985) Human stratum corneum polar lipids and desquamation.Arch. Derm. Res. 277: 284–287.

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Correspondence to Kyung Ho Row.

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Hong, S.P., Lee, C.H., Kim, S.K. et al. Mobile phase compositions for ceramide III by normal phase high performance liquid chromatography. Biotechnol Bioproc E 9, 47–51 (2004). https://doi.org/10.1007/BF02949321

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Keywords

  • Saccharomyces cerevisiae
  • cermaide III
  • mobile phase
  • NP-HPLC
  • ELSD