刺参性腺中6种磷脂含量的HPLC-ELSD分析

Detection of six phospholipid classes in gonad of sea cucumber Apostichopus japonicus by HPLC-ELSD

为了研究刺参Apostichopus japonicus性腺的磷脂组成,对刺参性腺中的磷脂进行分离纯化,利用高效液相色谱-蒸发光散射检测器,以ChromolithPerformance-Si正相整体硅胶柱(100 mm×4.6 mm)为固定相,使用正己烷(含0.04%三乙胺)、异丙醇和13%乙酸为流动相进行三元梯度洗脱,流速为1.5 m L/min,柱温为25℃,整个洗脱程序仅需24 min,蒸发光检测器温度为45℃,雾化气(空气)流速为1.7 L/min,在此条件下,对所提取的磷脂酰乙醇胺(PE)、磷脂酰肌醇(PI)、磷脂酰丝氨酸(PS)、磷脂酰胆碱(PC)、鞘磷脂(SM)和溶血磷脂酰胆碱(LPC)进行测定。结果表明:刺参性腺磷脂中PC含量最高,为4.637 g/kg,其次为PE和PS,分别为1.884、0.847 g/kg。研究表明,用此方法测定刺参性腺中的磷脂快速、准确,各磷脂组分分离良好,重复性高,对其他磷脂样品的测定也具有重要的参考价值。

The phospholipids were isolated and purified in gonad of sea cucumber Apostichopus japonicus by a high- performance liquid chromatograph equipped with an evaporative light scattering detector （HPLC-ELSD） and by a Chromolith Performance-Si normal-phase silica （ 100 mm×4.6 mm i. d. ） column at 25℃. The phospholipids was separated by a mobile phase consisting of 13% CH3 COOH,2-propanol and hexane with 0.04% triethylamine （TEA） in 24 min elution program including column re- equilibration, with flow rate of 1.5 mL/min in mobile phase at an evaporation temperature of 45 ~C at ELSD, and nebulization gas was air at a flow rate of 1.7 L/min. The analysis of the phosphlipids by HPLC-ELSD revealed that phosphatidylethanolamine （PC） （4. 637 g/kg） was the maximal phospholipid class, followed by the contents of phosphatidylethanolamine （PE） （ 1. 884 g/kg） and phosphatidylserine （PS） （0. 847 g/kg）, indicating that HPLC-ELSD was an accurate and reproducible analytical method for the detection of phosphlipids including PE, phosphatidylinositol （ PI）, PS, PC, sphingomyelin （SM） and lysophosphatidylcholine （LPC） in gonad of the sea cucumber.