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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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.
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.
Rupeic, J. and V. Maric (1998) Isolation and chemical composition of the ceramide of theCandida lipolytica yeast.Chem. Phys. Lipids 91: 153–161.
Aida, E. C. and S. F. Anthony (2000) Current methods for the identification and quantitation of ceramides: An overview.Lipids 35: 937–945.
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.
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.
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.
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.
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.
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.
Raith, K., J. Darius, and R. H. H. Neubert (2000) Ceramide analysis utilizing gas chromatography-mass spectrometry.J. Chromatogr. A 876: 229–233.
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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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
- Saccharomyces cerevisiae
- cermaide III
- mobile phase