Ishikawa T T, Brazier J B, Steiner P M, Stewart L E, Gartside P S, Glueck C J
Lipids. 1976 Aug;11(8):628-33. doi: 10.1007/BF02532877.
To assess the limits of the heparin MnCl2 precipitation method for quantitation of alpha-lipoprotein cholesterol (C-HDL), effects of varying final McCl2 and heparin concentrations were studied, and the precipitation method was compared to preparative ultracentrifugation. In 65 parallel plasma aliquots, C-HDL (X+/-SE) determined by ultracentrifugation (54.3+/-1.8 mg/dl) correlated significantly (r=0.98, P less than 0.001) with the precipitation method (56.0+/-1.9 mg/dl). C-HDL by ultracentrifugal and precipitation methods were also similar in 16 subjects with triglycerides ranging from 150 to 312 mg/dl (41.4+/-2.6, 43.4+/-2.8, r=0.97, P less than .001). A constant amount of cholesterol in the supernatant was measured over a final McCl2 range of 0.046-0.23 M, and cholesterol values in the supernatant at final McCl2 concentrations of 0.046, 0.05, and 0.055 M did not differ from each other, P greater than 0.1. However, cholesterol levels in the supernatant at final MnCl2 concentration of 0.042 M differed from those at concentrations of 0.046, 0.05, and 0.055 M, P less than 0.05 and the amount of supernatant cholesterol increased as the final McCl2 concentration was reduced from 0.042 to 0.02 M. A constant amount of cholesterol in the supernatant was measured over a heparin concentration range of 92-734 USP units/ml. The final MnCl2 and heparin concentrations of 0.046 M and 184 USP units/ml, which are incorporated in widely used procedures, gave C-HDL values for the precipitation method which were in close agreement with the ultracentrifugal method. There is no evidence for a heparin-Mn++ precipitation of HDL and systematic underestimation of HDL by the precipitation method. However, the final MnCl2 concentration is very near the minimum required for accurate measurement of C-HDL. To preclude incomplete precipitation of low and very low density lipoproteins by insufficient manganese concentration, an increase of the manganese concentration should be considered.
为评估肝素 - 氯化锰沉淀法测定α - 脂蛋白胆固醇（C - HDL）的局限性，研究了不同最终氯化锰和肝素浓度的影响，并将沉淀法与制备性超速离心法进行比较。在65份平行的血浆样本中，通过超速离心法测定的C - HDL（X±SE）为（54.3±1.8mg/dl），与沉淀法测定值（56.0±1.9mg/dl）显著相关（r = 0.98，P＜0.001）。在16名甘油三酯水平为150至312mg/dl的受试者中，超速离心法和沉淀法测得的C - HDL也相似（分别为41.4±2.6、43.4±2.8，r = 0.97，P＜0.001）。在最终氯化锰浓度范围为0.046 - 0.23M时，上清液中胆固醇含量恒定，最终氯化锰浓度为0.046M、0.05M和0.055M时，上清液中的胆固醇值彼此无差异，P＞0.1。然而，最终氯化锰浓度为0.042M时上清液中的胆固醇水平与0.046M、0.05M和0.055M时不同，P＜0.05，且随着最终氯化锰浓度从0.042M降至0.02M，上清液中胆固醇含量增加。在肝素浓度范围为92 - 734USP单位/ml时，上清液中胆固醇含量恒定。广泛使用的方法中采用的最终氯化锰和肝素浓度分别为0.046M和184USP单位/ml，沉淀法测得的C - HDL值与超速离心法非常接近。没有证据表明HDL会发生肝素 - 锰离子沉淀，也没有证据表明沉淀法会系统性低估HDL。然而，最终氯化锰浓度非常接近准确测量C - HDL所需的最低浓度。为避免因锰浓度不足导致低密度和极低密度脂蛋白沉淀不完全，应考虑提高锰浓度。
