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We describe a kinetic method for assay of serum or plasma triglycerides, by use of an enzymatic hydrolysis and reaction sequence already described [Clin. Chem. 19, 476 (1973)]. The reaction is triggered by addition of lipase, at a time when free glycerol, or pyruvate (or both) are no longer present. In this method, therefore, there is no need for a blank glycerol assay. In the procedure, reagents are used that are available commercially in the form of stable, dry powders; the method for the preparation of the reagents has been changed to achieve improved stability and performance. Stability and recovery of added triglycerides are satisfactory.