Leiss O, Murawski U, Egge H
J Clin Chem Clin Biochem. 1979 Oct;17(10):619-25.
A method is described which allows the determination of phospholipids, free and esterified cholesterol, triglycerides and free fatty acids in lipoprotein fractions starting from 50 microliter of serum. Lipoproteins were separated by successive precipitation: VLDL with Heparin/Mg++, LDL with Dextran sulfate/Mg++ and finally HDL with Dextran sulfate/Mn++. Lipids extracted from the precipitated lipoproteins were determined gravimetrically and by densitometry after thin layer chromatography and charring (van Gent, C.M. (1968), Z. Anal. Chem. 236, 344--350; Egge, H. et al. (1970) Z. Klin. Chem. Klin. Biochem. 8, 488--491). The results obtained from the serum of 12 adult healthy persons were compared with those from lipoprotein fractions separated by preparative ultracentrifugation (Havel, R. J. et al. (1955) J. Clin. Invest. 34, 1345--1353). The distribution of lipids in beta-lipoproteins (d less than 1.063 g/ml) and HDL (1.063 less than d less than 1.21 g/ml) prepared by both methods showed good agreement. Some differences were observed between VLDL (d less than 1.006 g/ml) and VHDL (d greater than 1.21 g/ml) prepared either by precipitation or ultracentrifugation. Compared to the total lipid of the sera, recovery rates were 95--105%. Variation coefficients were in the range of 15--20% for VLDL lipids, 5--10% for LDL and HDL lipids and 10--15% for VHDL lipids. Gravimetrically determined total lipids had a variation coefficient of 4 and 6% for LDL and HDL respectively.
本文描述了一种方法,该方法能够从50微升血清开始测定脂蛋白组分中的磷脂、游离胆固醇和酯化胆固醇、甘油三酯以及游离脂肪酸。脂蛋白通过连续沉淀进行分离:用肝素/镁离子沉淀极低密度脂蛋白(VLDL),用硫酸葡聚糖/镁离子沉淀低密度脂蛋白(LDL),最后用硫酸葡聚糖/锰离子沉淀高密度脂蛋白(HDL)。沉淀的脂蛋白中提取的脂质通过重量法测定,并在薄层色谱和炭化后进行光密度测定(范根特,C.M.(1968年),《分析化学杂志》236卷,344 - 350页;埃格,H.等人(1970年),《临床化学与临床生物化学杂志》8卷,488 - 491页)。将12名成年健康人的血清所得结果与通过制备性超速离心分离的脂蛋白组分的结果进行比较(哈维尔,R.J.等人(1955年),《临床研究杂志》34卷,1345 - 1353页)。两种方法制备的β - 脂蛋白(密度小于1.063克/毫升)和HDL(1.063小于密度小于1.21克/毫升)中的脂质分布显示出良好的一致性。通过沉淀或超速离心制备的VLDL(密度小于1.006克/毫升)和VHDL(密度大于1.21克/毫升)之间观察到一些差异。与血清总脂质相比,回收率为95 - 105%。极低密度脂蛋白脂质的变异系数在15 - 20%范围内,低密度脂蛋白和高密度脂蛋白脂质的变异系数在5 - 10%范围内,VHDL脂质的变异系数在10 - 15%范围内。重量法测定的低密度脂蛋白和高密度脂蛋白总脂质的变异系数分别为4%和6%。