Determination of Mobile Phase Compositons for Separation of Ceramide-Iii on NP-HPLC

초록

We experimentally determined the mobile phase composition to separate ceramide-III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol, n-hexane/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/n-butanol, the first mobile phase composition, 96/4 vol.%, was step-increased to 54/46 vol. % at 3 min. While in the binary mobile phase, the retention time of ceramide-III was 7.73 min and its number of theoretical plates was 17800, those were 4.11 min and 9500, respectively in the ternary mobile phase, where the mobile phase composition of n-hexane/IPA/methanol, 85/7/8 vol. %, was step-increased to 75/10/15 vol.% at 3 min. In both cases, the ceramide-III was well resolved.