State Key Laboratory for Bioactive Substances and Functions of Natural Medicines & Chinese Academy of Medical Sciences & Peking Union Medical College, PR China.
Chem Phys Lipids. 2012 Feb;165(2):133-41. doi: 10.1016/j.chemphyslip.2011.11.010. Epub 2011 Dec 1.
A simple and sensitive method to determine lipoprotein and lipids profiles in micro-liter scale individual serum sample is not presently available. Traditional lipoprotein separation techniques either by ultra-centrifugation or by liquid chromatography methods have their disadvantages in both lipoprotein separation and lipids component quantification. In this study we used small volume needing size-exclusion fast protein liquid chromatography to separate different lipoprotein subclasses in 50μL serum. And lipids contents, such as cholesterol, cholesterol ester and triacylglycerol, were measured by using two different fluorescence-based lipid detection methods. With this method, very low density lipoprotein, low density lipoprotein and high density lipoprotein could be easily separated, and follow-up lipid detection was completed by simple kinds of reactions. Serum lipoprotein and lipids profiling from C57BL/6 mice (n=5) and human (n=5) were analyzed. The elution profiles of five individuals were highly reproducible, and there were lipoprotein and lipids distribution variations between C57BL/6 mice and human beings. In conclusion, this method which combined small volume needing size-exclusion fast protein liquid chromatography and fluorescence-based lipids measurement, provided a simple, efficient, integrity and reproducible procedure for determining serum lipoprotein and lipids profiles in micro-liter scale levels. It becomes possible that determination of lipoprotein profiles and gaining information of lipids in different lipoproteins can be accomplished simultaneously.
目前尚无在微升级别个体血清样本中简单且灵敏地测定脂蛋白和脂质谱的方法。传统的脂蛋白分离技术，无论是超速离心法还是液相色谱法，在脂蛋白分离和脂质成分定量方面都存在缺点。在本研究中，我们使用小体积需求的分子筛快速蛋白液相色谱法在 50μL 血清中分离不同的脂蛋白亚类。并使用两种不同的基于荧光的脂质检测方法来测量胆固醇、胆固醇酯和三酰甘油等脂质含量。使用这种方法，可以轻松分离极低密度脂蛋白、低密度脂蛋白和高密度脂蛋白，并且可以通过简单的反应完成后续的脂质检测。对 C57BL/6 小鼠（n=5）和人类（n=5）的血清脂蛋白和脂质谱进行了分析。五个人的洗脱图谱具有高度重现性，并且 C57BL/6 小鼠和人类之间存在脂蛋白和脂质分布的差异。总之，这种结合了小体积需求的分子筛快速蛋白液相色谱和基于荧光的脂质检测的方法，为在微升级别水平上确定血清脂蛋白和脂质谱提供了一种简单、高效、完整和可重复的程序。同时测定脂蛋白谱并获得不同脂蛋白中脂质的信息成为可能。
