Nondestructive quantification of neutral lipids by thin-layer chromatography and laser-fluorescent scanning: Suitable methods for "lipidome" analysis
Nondestructive quantification of neutral lipids by thin-layer chromatography and laser-fluorescent scanning: Suitable methods for "lipidome" analysis
复制标题
DOI：
10.1006/bbrc.2001.4404
复制
发表时间：
2001-03-02
影响因子：
3.1
通讯作者：
Moriyama, T
中科院分区：
文献类型：
--
作者：
Kishimoto, K;Urade, R;Moriyama, T
采用薄层色谱和高效荧光扫描技术，建立了一种简便的中性脂质分离定量方法。采用双显影薄层色谱法分离中性脂类，罗丹明6G和激光激发荧光扫描仪检测。应用脂质量与扫描强度体积呈剂量依赖关系。通过将未知样品的条带强度与真实标准品的稀释曲线进行比较，确定每个中性脂带的质量。经染色喷雾TLC扫描后，提取的甘油三酯（TG）可用气相色谱法测定酰基链种类。用这种方法定量测定了小鼠肝脏中TG的含量，发现TG的总质量与酶法测定的总质量具有一定的相关性。我们的方法应该为“脂质组”分析提供基本技术，旨在确定和比较生物样品中存在的总脂类和质量。(C) 2001学术出版社。
A simple method for separation and quantification of neutral lipids was developed using thin-layer chromatography (TLC) and high-performance fluorescent scanning. Neutral Lipid classes were separated using the double-developing TLC method and detected by rhodamine 6G and a laser-excited fluorescent scanner. The amount of lipids applied correlated with scanned intensity volume in a dose-dependent manner. The mass of each neutral lipid band was determined by comparing band intensities of unknown samples to dilution curves of authentic standards. After scanning the dye-sprayed TLC, acyl chain species of triglyceride (TG) extracted from TLC could be determined by gas chromatography. Using this method, we quantified the amounts of TG in mouse liver and found that the measured total mass of TG correlated with that obtained by enzymatic methods. Our method should provide the basic technique for "lipidome" analysis, designed to determine and compare total lipid classes and mass present in biological samples. (C) 2001 Academic Press.