Ceramide III was prepared by the cultivation of Saccharomyces 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 dete...
Ceramide III was prepared by the cultivation of Saccharomyces 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/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-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 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-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.
Ceramide III was prepared by the cultivation of Saccharomyces 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/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-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 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-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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제안 방법
The purpose of this work is to find the optimum condi tion of mobile phase composition to analyze ceramide III from Saccharomyces cerevisiae based on the peak area of ceramide III using NP-HPLC with ELSD.
대상 데이터
Korea) installed on an HP Vectra 500 PC. The detection system was ELSD from Alltech (Deerfield, USA).
6 mm. The packing material was OP Sil-51002546 (Daejeon, Korea).
참고문헌 (12)
Biochim. Biophys. Acta J. A. Bouwstra 1300 177 1996 10.1016/0005-2760(96)00006-9 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.
Anal. Chim. Acta K. Raith 403 295 2000 10.1016/S0003-2670(99)00661-3 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.
Chem. Phys. Lipids J. Rupeic 91 153 1998 10.1016/S0009-3084(97)00106-0 Rupeic, J. and V. Maric (1998) Isolation and chemical composition of the ceramide of theCandida lipolytica yeast.Chem. Phys. Lipids 91: 153-161.
Lipids E. C. Aida 35 937 2000 10.1007/s11745-000-0603-1 Aida, E. C. and S. F. Anthony (2000) Current methods for the identification and quantitation of ceramides: An overview.Lipids 35: 937-945.
Appl. Microbiol. Biotechnol. J. Rupeic 50 583 1998 10.1007/s002530051339 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.
Biochim. Biophys. Acta T. Gildenast 1346 69 1997 10.1016/S0005-2760(97)00019-2 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.
J. Chromatogr. A K. Gaudin 883 211 2000 10.1016/S0021-9673(00)00445-3 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.
J. High Resol. Chromatogr. P. Molander 23 653 2000 10.1002/1521-4168(20001101)23:11<653::AID-JHRC653>3.0.CO;2-A 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.
J. Biol. Chem. G. Wells 273 7235 1998 10.1074/jbc.273.13.7235 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.
Analy. Biochem. J. M. Thomas 276 242 1999 10.1006/abio.1999.4354 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.
J. Chromatogr. A K. Raith 876 229 2000 10.1016/S0021-9673(00)00144-8 Raith, K., J. Darius, and R. H. H. Neubert (2000) Ceramide analysis utilizing gas chromatography-mass spectrometry.J. Chromatogr. A 876: 229-233.
Arch. Derm. Res. A. S. Long 277 284 1985 10.1007/BF00509081 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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