Effect of lyophilization on results of five enzymatic methods for cholesterol

We studied the effect of lyophilization of serum pools on the determination of cholesterol with the enzymatic methods used in five automated analyzers: SMAC (Technicon), the RA-1000 (Technicon), the aca (Du Pont), the TDx (Abbott), and the Ektachem 700P (Kodak). We prepared two serum pools: pool A (2.2 g/L) and pool B (1.9 g/L). We separated each pool into three groups for treatment by lyophilizing or freezing at -20 and -70 degrees C. We determined cholesterol by the above methods at regular intervals during the next 270 days. For all methods, the measured concentration of cholesterol was less for the lyophilized serum than for frozen (P less than 0.007 by the paired-sample t-test). After adjusting for the dilutional effects of reconstitution, the decrease in original value was 0.7% for the SMAC, 1.7% for the RA-1000, 5.7% for the Ektachem, 9.4% for the TDx, and 14.3% for the aca. Lyophilization of cholesterol standards or serum samples may hamper the effective standardization of enzymatic methods and may be a source of apparent inaccuracy (bias) among enzymatic methods.