蒸发光散射检测器在唾液脂质检测中的应用

Application of DAD and ELSD in Detection of Salivary Lipids

目的采用高效液相色谱技术-蒸发光散射检测器对唾液脂质进行分离,并比较蒸发光散射检测器与二极管阵列检测器在唾液脂质检测中脂质色谱行为的差异。方法收集100ml全唾液,提取脂质,采用正相色谱分离唾液中性脂及磷脂,分别用二极管阵列检测器与蒸发光散射检测器进行检测。结果分离出四种中性脂组分,分别为胆固醇、油酸、1,3-二棕榈酸甘油酯、三脂酰甘油酯、两种磷脂组分,分别为磷脂酰胆碱、磷脂酰乙醇胺。蒸发光散射检测器与二极管阵列检测器比较,基线稳定,分辨率高。结论对于痕量的唾液脂质,采用蒸发光散射检测器效果优于二极管阵列检测器。

Objective To separarate salivary lipids by high performance liquid chromatography (HPLC)-evaporation-light-scattering detector(ELSD) and to compare the behavier of lipids chromatography by diode array detector(DAD) and ELSD.Methods The study was conducted with 100ml whole salivary which were obtained and treated by the means of Slomiany B L.Lipids were extracted and purified by the means of Folch.After dried under N2,the extractions were dissolved in chlorform/methanol(2/1)for further separation.The separation was done with an Kromasil SIL column as analytical column,salivary neutral lipids were achieved with solvent mixture of hexane/isopropanol/0.5%ammonium hydroxide=98/1.5/0.5(v/v/v)and phospholipids were achieved with acetonitrile/methanol/0.1%phosphoric acid aqueous solution=50/50/1(v/v/v)as the mobile phase respectively.Tested two HPLC methods for salivary lipids analysis using DAD as well as ELSD.Cholesterol,oleic acid,1,3-dipalmitin,triglycerol phosphatidylethanolamine and phosphatidylcholine ware identified by authentics obtained in 99% purity from Sigma Chemical Co.(St.Louis,Mo.U.S.A).Results Compared to DAD,the baseline was stable and the sensitivity was high with ELSD.Conclusion ELSD is the better detector for testing salivary lipids.