Entezami A A, Venables B J, Daugherty K E
J Chromatogr. 1987 Jan 30;387:323-31. doi: 10.1016/s0021-9673(01)94535-2.
A high-efficiency silica gel, (type HLF) thin-layer chromatography plate (HETLC), linear high-performance thin-layer chromatography plate (HPTLC) and densitometry method has been devised in order to resolve the major lipid classes obtained from rat brain tissues. This methodology, which has largely overcome prior problems, enhances the opportunity for assessing the glycerophospholipid and glycolipid compositions of tissues. DEAE-Sephadex column chromatography was used to separate the crude lipids extract into neutral and acidic lipid fractions. The lipid fractions were then spotted on separate HPTLC and HETLC plates and chromatographed in one dimension using one solvent system. Quantitation was by in situ densitometry with the absolute quantity of the lipid classes determined from co-chromatographed standards. Sensitivity was increased by using cupric sulfate reagent, which was found to be more sensitive than the conventional cupric acetate reagent. This method is applicable to a broader separation of lipid classes and has improved sensitivity.
为了分离从大鼠脑组织中获得的主要脂质类别，设计了一种高效硅胶（HLF型）薄层色谱板（HETLC）、线性高效薄层色谱板（HPTLC）和密度测定法。这种方法在很大程度上克服了先前的问题，增加了评估组织甘油磷脂和糖脂组成的机会。使用DEAE-葡聚糖柱色谱将粗脂质提取物分离为中性和酸性脂质部分。然后将脂质部分点样在单独的HPTLC和HETLC板上，并使用一种溶剂系统进行一维色谱分离。通过原位密度测定法定量，脂质类别的绝对量由共色谱标准品确定。使用硫酸铜试剂提高了灵敏度，发现其比传统的醋酸铜试剂更灵敏。该方法适用于更广泛的脂质类别分离，并且提高了灵敏度。
