Institute of Geriatrics Beijing Hospital, Beijing 100730, China.
J Lipid Res. 2011 Feb;52(2):383-8. doi: 10.1194/jlr.D008979. Epub 2010 Nov 1.
Simple and precise methods for LDL-cholesterol (LDL-C) and HDL-cholesterol (HDL-C) measurements are essential for assessment of cardiovascular disease (CVD) risks and for lipid and lipoprotein studies. We report here an ultracentrifugation (UC) and HPLC method that requires substantially less specimen volume and provides the necessary reliability and throughput required by large-volume, high-quality research and clinical studies. 2-Mercaptoethanol (ME) was used to dissociate serum lipoprotein [a] (Lp[a]) into apolipoprotein [a] and Lp[a] remnant (Lp[a-]) and eliminated the contamination of Lp[a] in HDL separated by UC. Serum aliquots were centrifuged at a density of 1.006 kg/l for the separation of HDL plus LDL, and in the presence of ME at a density of 1.063 kg/l for the separation of HDL. Cholesterol concentrations of the bottom fractions were analyzed by HPLC. LDL-C and HDL-C determined using this method were equivalent to those with β-quantification and the designated comparison method of the Centers for Disease Control. The total coefficient of variations for LDL-C and HDL-C were 0.65-1.12% and 0.96-2.07%, respectively. This method requires a small amount of specimen and is easy to operate. This method may be used in research or in clinical laboratories where precise and specific lipoprotein cholesterol analysis is needed.
用于 LDL-胆固醇（LDL-C）和 HDL-胆固醇（HDL-C）测量的简单而精确的方法对于评估心血管疾病（CVD）风险以及脂质和脂蛋白研究至关重要。我们在此报告一种超速离心（UC）和 HPLC 方法，该方法需要的标本量大大减少，并提供了大容量、高质量研究和临床研究所需的必要可靠性和通量。使用 2-巯基乙醇（ME）将血清脂蛋白[a]（Lp[a]）解离为载脂蛋白[a]和 Lp[a]残基（Lp[a-]），并消除了 UC 分离的 HDL 中 Lp[a]的污染。血清等分试样以 1.006 kg/l 的密度离心以分离 HDL 和 LDL，并且在 ME 的存在下以 1.063 kg/l 的密度分离 HDL。底部馏分的胆固醇浓度通过 HPLC 进行分析。使用该方法测定的 LDL-C 和 HDL-C 与β定量和疾病控制中心指定的比较方法相当。LDL-C 和 HDL-C 的总变异系数分别为 0.65-1.12%和 0.96-2.07%。该方法需要少量标本，操作简单。该方法可用于需要精确和特定脂蛋白胆固醇分析的研究或临床实验室。
