Enzymatic method for determination of CO2 in serum

Abstract

We describe an enzymatic method, requiring only 10 mul of serum, for determining CO2 as bicarbonate or dissolved gas. Phosphoenolpyruvate carboxylase catalyzes the reaction of HCO3- with phosphoenolypyruvate to give oxalacetate. The resulting NADH, in the presence of malate dehydrogenase, is oxidized to NAD+, and the decrease in absorbance at 340 nm is directly proportional to the amount of CO2 present in the sample. Reaction is complete in 3 to 6 min under assay conditions, and is linearly related to CO2 concentrations between 8 and 65 mmol/liter. Analytical recovery is 95-110% (average, 101%). Two laboratories compared values obtained by continuous-flow analysis. The resulting correlation coefficients were 0.966 and 0.987, values for the t-test were t(paired) equals 0.473 and t(paired) equals 0.334, and average day-to-day precisions (three concentrations) were 3.9% and 4.2%.