In the clinical laboratory, we routinely measure the cholesterol content of high-density lipoprotein and low-density lipoprotein particles and not the apolipoproteins on the particles or the number of particles. Proprietary detergents and reagents are used in assays for HDL-C and LDL-C to separate lipoproteins, allowing the cholesterol content of specific lipoproteins to be measured. For example, HDL-C is commonly measured using a solution of dextran sulfate and magnesium to selectively precipitate HDL from the other lipoproteins present in the sample. Once isolated, the HDL particles are 'dissolved' and the amount of cholesterol in them is determined photometrically using a color-producing enzyme reaction. LDL-C can be measured directly or can be estimated using the HDL-C, triglycerides and total cholesterol (TC) values. The Friedewald formula is often used to calculate LDL:
LDL-C = TC - (HDL-C)- (Triglycerides/5).
The important point to consider here is that traditional LDL-C and HDL-C measurements only tell us how much cholesterol is associated with each lipoprotein particle class. We are now learning that the number and size of the particles are important as well. The number of LDL particles appears to be more strongly predictive of cardiovascular disease than the LDL-C content, and small dense LDL are known to be more atherogenic than larger, less dense LDL particles.