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

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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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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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  • Saccharomyces cerevisiae
  • cermaide III
  • mobile phase
  • ELSD