高效液相色谱-蒸发光散射法分析海参脑苷脂和神经酰胺的含量

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

中文关键词:  高效液相色谱法  蒸发光散射检测法  神经酰胺  脑苷脂  海参

英文关键词:high performance liquid chromatography  evaporative light-scattering detection  ceramides  cerebrosides  sea cucumber

中文摘要:

      目的 建立了一种高效液相色谱-蒸发光散射检测方法(high performance liquid chromatography coupled with evaporative light-scattering detection, HPLC-ELSD)同时分析海参中脑苷脂和神经酰胺含量的方法。方法 以TSKgel CN-80Ts柱为分离柱, 正己烷-异丙醇-二氯甲烷-甲醇为流动相, 采用梯度洗脱方式, 流速1.0 mL/min, 漂移管温度50 ℃, 氮气流速2.4 L/min。结果 在优化条件下, 18 min内完成分离, 脑苷脂的线性范围为2.5~250 μg/mL, 检出限为0.5 μg/mL(S/N=3), 相关系数R>0.99, 神经酰胺的线性范围为5~150 μg/mL, 检出限为0.05 μg/mL(S/N=3), 相关系数R>0.99, 方法稳定性良好; 不同种海参中脑苷脂和神经酰胺含量存在较为明显的差异, 含量范围分别为0~4.44 mg/g和0.24~2.21 mg/g。其中, 猪参的脑苷脂含量最高为4.44 mg/g, 而白底靴参的脑苷脂未能检出。阿拉斯加红参的神经酰胺含量最高为2.21 mg/g, 而海地瓜神经酰胺含量最少。结论 所建立的方法灵敏、准确、快速, 可应用于食品中脑苷脂和神经酰胺的同时定量分析。

英文摘要:

      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-80Ts column by using n-hexane-isopropanol-dichloromethane-methanol as the mobile phase at a flow rate of 1.0 mL/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/mL, and the limit of detection was 0.5 μg/mL with a regression correlation coefficient of 0.99. Meanwhile, the linear range of the established ceramides calibration curve was 5~150 μg/mL, and the limit of detection was 0.05 μg/mL 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.