Yasugi E, Oshima M
Division of Clinical Biochemistry, National Medical Center, Tokyo, Japan.
Biochim Biophys Acta. 1994 Feb 10;1211(1):107-13. doi: 10.1016/0005-2760(94)90144-9.
Sequential microanalyses of free dolichol, dolichyl fatty acid ester and dolichyl phosphate in human serum were made. To determine the level of each dolichol, samples were pretreated using three different methods prior to fluorescent derivatization. To estimate the concentrations of free dolichol, samples were added to alkaline methanol and kept at room temperature for 1 h. In case of dolichyl fatty acid ester, samples were saponified at 100 degrees C for 1 h. To estimate dolichyl phosphate, saponified lipid extracts were treated with acid phosphatase. Each pretreated dolichol was reacted with anthracene-9-carboxylic acid and amounts of 9-anthroyl derivatives were determined fluorometrically by HPLC. This method is simple and three types of dolichols can be estimated using the same HPLC system. This analysis is also sufficiently sensitive for measurement of serum dolichol levels. The contents of free dolichol, dolichyl fatty acid ester and dolichyl phosphate in human serum were found to be 44.9 +/- 10.5 ng/ml (n = 32), 76.4 +/- 24.2 ng/ml (n = 32) and 43.5 +/- 15.1 ng/ml (n = 13), respectively. These levels had apparently no correlation to age or serum total cholesterols. A linear correlation between dolichols and high-density lipoprotein cholesterols reflects the fact that the dolichols are associated with the high-density lipoprotein fraction.
对人血清中的游离多萜醇、多萜醇脂肪酸酯和多萜醇磷酸酯进行了连续微量分析。为了测定每种多萜醇的水平，在荧光衍生化之前，使用三种不同方法对样品进行预处理。为了估计游离多萜醇的浓度，将样品加入碱性甲醇中并在室温下保存1小时。对于多萜醇脂肪酸酯，样品在100℃皂化1小时。为了估计多萜醇磷酸酯，皂化的脂质提取物用酸性磷酸酶处理。每种预处理的多萜醇与蒽-9-羧酸反应，并通过高效液相色谱法荧光测定9-蒽酰衍生物的量。该方法简单，并且可以使用相同的高效液相色谱系统估计三种类型的多萜醇。该分析对于测量血清多萜醇水平也具有足够的灵敏度。发现人血清中游离多萜醇、多萜醇脂肪酸酯和多萜醇磷酸酯的含量分别为44.9±10.5 ng/ml（n = 32）、76.4±24.2 ng/ml（n = 32）和43.5±15.1 ng/ml（n = 13）。这些水平显然与年龄或血清总胆固醇无关。多萜醇与高密度脂蛋白胆固醇之间的线性相关性反映了多萜醇与高密度脂蛋白部分相关的事实。
