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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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Authors and Affiliations

  1. Center for Advanced Bioseparation Technology, Department of Chemical Engineering, Inha University, 402-751, Incheon, Korea

    Seung Pyo Hong, Chong Ho Lee & Kyung Ho Row

  2. Department of Biological Engineering, Inha University, 402-751, Incheon, Korea

    Se Kyung Kim & Hyun Shik Yun

  3. Department of Chemical Engineering, Chosun University, 501-759, Kwangju, Korea

    Jung Heon Lee

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  1. Seung Pyo Hong
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  2. Chong Ho Lee
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  3. Se Kyung Kim
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  4. Hyun Shik Yun
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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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  • DOI: https://doi.org/10.1007/BF02949321

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