Causes and elimination of erratic blanks in enzymatic metabolite assays involving the use of NAD+ in alkaline hydrazine buffers: Improved conditions for the assay of l-glutamate, l-lactate, and other metabolites

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Why is it important?

These high pHassays use hydrazine to trap keto products e.g. pyruvate. A student practical on lactate production gave wildly variable results. It emerged that without any lactate present the hydrazine reacts directly with NAD+ to give an interfering and variable blank. The variability is due to metal catalysis and is largely eliminated by EDTA.