Whole blood L-lactate assay by a new differential pH method: application to metabolic investigations

We propose a new quantitative method for L-lactate assay in whole blood, based on the measurement of pH variation caused by specific and irreversible oxidation of L-lactate to pyruvate in the presence of an electron acceptor (hexacyanoferrate) and of the enzyme cytochrome b2 (EC 1.1.2.3.). No sample pretreatment is needed; the method is simple and fast (1.5 min/analysis) and requires 10 microliters whole blood per assay. Linearity is confirmed up to 20 mmol/l L-lactate. Within-day and between-day variability was (as C.V.) 3.6% and 8.1% for blood lactate 1.3 and 1.0 mmol/l, respectively. The results by the present method correlate well with those from two reference methods (test method vs a lactate sensor based method: r = 0.996; test method vs a spectrophotometric method: r = 0.987). An application of the present method to the continuous monitoring of L-lactate in patients after combined kidney and pancreas transplantation, under conditions of euglycemic hyperinsulinemia and hyperglycemic hyperinsulinemia is reported. We conclude that the method is simple and reproducible and can be employed to measure whole blood lactate concentration continuously both in clinical protocols and in basic research.