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Simultaneous quantitative analysis of cerebrosides and ceramides from sea cucumbers by high performance liquid chromatography coupled with evaporative light-scattering detection coupled with evaporative light-scattering detection

Objective To establish a high performance liquid chromatography coupled with evaporative light-scattering detection(HPLC-ELSD) method for analyzing the contents of cerebrosides and ceramides simultaneously in sea cucumbers. Methods The separation was performed on a TSK gel CN-80 Ts column by using n-hexane-isopropanol-dichloromethane-methanol as the mobile phase at a flow rate of 1.0 m L/min. The temperature of drift tube was kept at 50 ℃ and the flow rate of nitrogen was 2.4 L/min. Seventeen sea cucumber species were selected as experimental subjects. Results Under optimal conditions, effective separation of cerebrosides and ceramides was completed within 18 min. The linear range of the established cerebrosides calibration curve was 2.5~250 μg/m L, and the limit of detection was 0.5 μg/m L with a regression correlation coefficient of 0.99. Meanwhile, the linear range of the established ceramides calibration curve was 5~150 μg/m L, and the limit of detection was 0.05 μg/m L with a regression correlation coefficient of 0.99. There were obvious difference in cerebrosides and ceramides among these sea cucumber species. The cerebrosides content of Bohadschia marmorata was the highest, which was up to 4.44 mg/g, in contrast, that of Actinopyga mauritiana could not be detected. As for ceramides, the content of Parastichopus californicus was the highest, which was up to 2.21 mg/g, in contrast, Acaudina molpadioides had the lowest content. Conclusion The established method can provide essential data for development and utilization of sea cucumbers, which is sensitive, accurate and fast.