Preparative high-performance liquid chromatography purification of polyunsaturated phospholipids and characterization using ultraviolet derivative spectroscopy.
Preparative high-performance liquid chromatography purification of polyunsaturated phospholipids and characterization using ultraviolet derivative spectroscopy.
复制标题
多不饱和磷脂的制备型高效液相色谱纯化和紫外导数光谱表征。
DOI：
10.1016/0003-2697(90)90541-g
复制
发表时间：
1990
影响因子：
2.9
通讯作者：
Dratz,EA
中科院分区：
文献类型：
--
作者：
Holte,LL;vanKuijk,FJ;Dratz,EA
一种制备反相高效液相色谱系统利用等密度流动相纯化高达10毫克的磷脂量被描述。建立了从完整的母体脂质中分离多不饱和磷脂氧化产物的方法。该方法适用于制备级的磷脂酰胆碱和磷脂酰乙醇胺，以及分析级的磷脂酰丝氨酸和磷脂酸。完整磷脂和氧化磷脂在206nm处进行吸光度监测。氧化产物在234 nm处同时被监测，完整的磷脂只有非常轻微的末端吸收。二阶导数紫外光谱被证明是非常有用的，以确定存在或验证不氧化产物的磷脂样品。对于含磷脂的自氧化二十二碳六烯酸，反式、反式（t,t）异构体的最大吸光度为237 nm，顺式、反式（c,t）异构体的最大吸光度为246 nm。同样，五类三元氧化产物立体异构体在269 ~ 292 nm范围内具有不同的吸光度最大值。
A preparative reversed-phase HPLC system utilizing an isocratic mobile phase to purify up to 10-mg quantities of phospholipids is described. The method was developed to separate oxidation products of polyunsaturated phospholipids from intact, parent lipids. The method is useful for phosphatidylcholine and phosphatidylethanolamine on a preparative scale and for phosphatidylserine and phosphatidic acid on an analytical scale. Both intact phospholipids and oxidized phospholipids were monitored by absorbance at 206 nm. The oxidation products were simultaneously monitored at 234 nm where the intact phospholipids have only a very slight end absorption. Second-derivative uv spectroscopy proved to be extremely useful to identify the presence or to verify the absence of oxidation products in phospholipid samples. For autoxidized docosahexaenoic acid containing phospholipids, the absorbance maximum of dience oxidation products is 237 nm for the trans,trans (t,t) isomer and 246 nm for the cis,trans (c,t) isomers. Similarly, five classes of trience oxidation product stereoisomers have distinct absorbance maxima detected by second-derivative spectroscopy ranging from 269 to 292 nm.