Simplified methods for measuring cholesterol concentrations of high-density lipoprotein subclasses in serum compared.
We used a combination of heparin micro-affinity column chromatography and heparin-Mn2+/dextran sulfate (DS) precipitation procedures to measure directly the total high-density lipoprotein (HDL)-cholesterol and HDL3-cholesterol in serum. The value for HDL2-cholesterol was obtained by subtracting the value for HDL3-cholesterol from that for total HDL-cholesterol. Results of this methodology and of the original heparin-Mn2+/DS double-precipitation method were compared with those of preparative ultracentrifugation. Concentrations of HDL- and HDL2-cholesterol by the original double-precipitation method were respectively 26 and 33 mg/L lower than by the ultracentrifugation method (p less than 0.001). Corresponding values by the micro-affinity/precipitation method were 36 and 29 mg/L higher than by the ultracentrifugation method (p less than 0.001). The values for HDL3-cholesterol by ultracentrifugation and by precipitation differed only by 7 mg/L (p greater than 0.05). Micro-affinity/precipitation and double-precipitation results both correlated well with those by ultracentrifugation (r = 0.84 to 0.94); the y-intercept of the comparison with the micro-affinity/precipitation method was close to zero. Use of the micro-affinity/precipitation method with samples from black or white adolescents showed that the blacks had higher concentrations of HDL2-(p less than 0.01) and HDL3-cholesterol (p less than 0.02) than did the whites.
D000293:Adolescent; D001055:Apolipoproteins B / Q000032:analysis; D011232:Chemical Precipitation / Q000379:methods; D002712:Chlorides; D002784:Cholesterol / Q000032:analysis*; D008076:Cholesterol, HDL / Q000032:analysis*; D002846:Chromatography, Affinity; D016264:Dextran Sulfate; D003911:Dextrans; D006493:Heparin; D006801:Humans; D008075:Lipoproteins, HDL / Q000097:blood* / Q000145:classification; D008345:Manganese; D017895:Manganese Compounds; D044469:Racial Groups; D014461:Ultracentrifugation
D001055:Apolipoproteins B; D002712:Chlorides; D008076:Cholesterol, HDL; D003911:Dextrans; D008075:Lipoproteins, HDL; D017895:Manganese Compounds; D008345:Manganese; D006493:Heparin; D016264:Dextran Sulfate; D002784:Cholesterol; C025340:manganese chloride