Assay instrument-dependent matrix effects in standardization of cholesterol measurements

Human serum-based frozen reference materials have been used by the Centers for Disease Control and Prevention (CDC)-National Heart, Lung and Blood Institute Lipid Standardization Program to improve the precision and accuracy of blood cholesterol measurements. Occasionally, laboratories in the program have had problems obtaining results for patients' fresh serum samples equivalent to those obtained with frozen CDC standardization pools. This incompatibility of sample, reagent, instrument, and assay characteristics has been labeled broadly as a "matrix effect," which usually is attributed to unknown characteristics of the processed pool material. In this study we showed that a large negative bias obtained with CDC pools was attributable to use of the sample blank mode on the Cobas-Bio analyzer. However, under the same conditions, fresh patients' serum samples were analyzed accurately. The use of a blank absorbance immediately after mixing sample and reagents (the "autoblank" mode) allowed the instrument to accurately analyze both fresh serum samples and CDC standardization pools and thus allowed the documentation of traceability of the cholesterol measurements to the National Reference System for Cholesterol.