Evaluation with the centrifugal fast analyzer of a chemical activation procedure for creatine kinase MB isoenzyme

The differential activation method for the determination of the "myocardial" isoenzyme of creatine kinase (MB) is based on the computed difference in activity of serum aliquots activated by the combination of dithiothreitol and glutathione (skeletal plus myocardial creatine kinase) and by glutathione alone (skeletal creatine kinase). The unique ability of the Centrifugal Fast Analyzer to perform analyses in parallel is used to precisely measure (CV = 0.1-1.5%) the activity of the two chemically activated aliquots. Statistical considerations concerning the reliability of this difference estimation are discussed with respect to both the precision of the rate measurements and to the relative amount of isoenzyme MB present. Another potential source of error in the analysis of the two differently activated aliquots, namely variation in lag phases, is circumvented by use of a linear-search FOCAL software package. This program searches the data for a linear segment of maximum slope, automatically rejecting those data that appear in lag or depletion regions of the curve representing the progress of the reaction. Correspondence of the results obtained with those of comparison techniques (chromatography and electrophoresis) are discussed.