Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Mobile Phase Compositions for Ceramide III by Normal Phase High Performance Liquid Chromatography

  • Hong, Seung-Pyo (Center for Advanced Bioseparation Technology, Department of Chemical Engineering) ;
  • Lee, Chong-Ho (Center for Advanced Bioseparation Technology, Department of Chemical Engineering) ;
  • Kim, Se-Kyung (Department of Biological Engineering, Inha University) ;
  • Yun, Hyun-Shik (Department of Biological Engineering, Inha University) ;
  • Lee, Jung-Heon (Department of Chemical Engineering, Chosun University) ;
  • Row, Kyung-Ho (Center for Advanced Bioseparation Technology, Department of Chemical Engineering)
  • Published : 2004.01.01
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Abstract

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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References

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