Standardization of serum cholesterol assays by use of serum calibrators and direct addition of Liebermann-Burchard reagent.

Abstract

Serum cholesterol concentrations of subjects in epidemiological studies were measured after direct addition of Liebermann-Burchard reagent; results were calibrated with human serum pools assayed according to Abell et al. (J. Biol. Chem. 195:357-366, 1952). Accuracy and precision were monitored for six years by analysis of internal-control pools and blind external-control pools. For various internal-control pools, the imprecision (CV) of the long-term averages of run means ranged from 0.5 to 0.9%. The within-run CV for internal control and patients' sera was about 1%. For blind control sera with different concentrations (provided by the Centers for Disease Control, Atlanta, GA, over the same period), the average difference per three-month period between the values found and the target values was usually between -0.5% and +0.7% for medium-concentration pools and between -2% and +2% for low- and high-concentration pools (extreme values: -2.4% and +2.5%). The CV per three-month period ranged from 0.6 to 2.7%. Sera from subjects on diets of high or low linoleic acid content were analyzed to study the effect of the fatty acid portion of serum cholesterol esters; the differences between values obtained with the comparison method and the direct method was insignificant on both diets. We conclude that the use of serum calibrators eliminates the bias inherent in the direct method.