Conlon D R, Blankstein L A, Pasakarnis P A, Steinberg C M, D'Amelio J E
Clin Chem. 1979 Nov;25(11):1965-9.
We have developed a procedure for the determination of high-density lipoprotein cholesterol by agarose gel electrophoresis. Only 2 micro L of sample was applied to the gel. After electrophoresis at 90 V for 35 min, an enzymatic cholesterol reagent was applied. After a 30-min incubation, the high-density lipoprotein cholesterol was quantified by densitometry. Precision for this measurment approaches that reported for the heparin-manganese/Abell-Kendall method (Clin. Chem, 25: 596--609, 1979). We evaluated accuracy by comparing high-density lipoprotein cholesterol concentration measured by electrophoresis to that determined in the Framingham Heart Study procedure (J. Biol. Chem. 195: 357, 1952). The resulting correlation was excellent. By the paired Student's t-test, there was no significant difference between the two methods. The proposed method gives a linear standard curve when the concentration of total cholesterol is between 1.0 and 3.5 g/L. By accurate quantitation of high-density lipoprotein cholesterol, agarose gel electrophoresis can aid in assessment of coronary heart disease risk for a large segment of the population.
我们已经开发出一种通过琼脂糖凝胶电泳测定高密度脂蛋白胆固醇的方法。仅将2微升样品加样到凝胶上。在90伏电压下电泳35分钟后，加入一种酶法胆固醇试剂。孵育30分钟后，通过光密度法对高密度脂蛋白胆固醇进行定量。该测量方法的精密度接近肝素-锰/阿贝尔-肯德尔法（《临床化学》，25: 596 - 609, 1979）所报道的精密度。我们通过比较电泳法测得的高密度脂蛋白胆固醇浓度与在弗雷明汉心脏研究程序（《生物化学杂志》，195: 357, 1952）中测定的浓度来评估准确性。所得相关性极佳。通过配对学生t检验，两种方法之间无显著差异。当总胆固醇浓度在1.0至3.5克/升之间时，所提出的方法给出线性标准曲线。通过对高密度脂蛋白胆固醇的准确定量，琼脂糖凝胶电泳有助于评估很大一部分人群的冠心病风险。
