Cyclic Enzymatic Determination of L-Lactate by Differential pH Measurement.

Abstract An electrochemical procedure for the analysis of L-lactate based on the lactate dehydrogenase/lactate oxidase reaction cycle has been developed and applied in whole blood and wine. The reaction of lactate with lactate oxidase produces pyruvate, which is the substrate, at physiological pH, of the enzyme lactate dehydrogenase. This enzyme, in the presence of NADH, converts pyruvate to lactate consuming H+ An automated software controlled differential pHmeter was used for H+ detection and the ΔpH measured was correlated to the lactate concentration present in the sample. Lactate was measured in the range 0.4–20 mmol/L with a detection limit of 0.2 mmol/L. Recovery studies have been carried out to evaluate the effect of blood and wine matrices on lactate determination. The recovery was between 94 and 104% in blood and 93 and 104% in wine. Analysis of lactate in blood and wine samples has been carried out. Comparison with a reference spectrophotometric procedure has been reported and discussed.

[1]  J. Raba,et al.  On-line enzymatic amplification by substrate cycling in a dual bioreactor with rotation and amperometric detection. , 1994, Analytical biochemistry.

[2]  G. Palleschi,et al.  Bioelectrochemical determination of lactic and malic acids in wine. , 1994, Talanta.

[3]  G. Palleschi,et al.  Amperometric biosensor for determination of lactate in sweat , 1993 .

[4]  A. Mosca,et al.  Rapid determination of erythrocyte pyruvate kinase activity. , 1993, Clinical chemistry.

[5]  A. Skillen,et al.  Amperometric enzyme electrode for NADH detection employing co-immobilized lactate dehydrogenase and lactate oxidase , 1992 .

[6]  E H Hansen,et al.  Optimization of flow-injection systems for determination of substrates by means of enzyme amplification reactions and chemiluminescence detection. , 1991, Talanta.

[7]  M. Tabata,et al.  Determinations of lactate and lactate dehydrogenase activity in serum with the flow injection analysis system involving immobilized enzyme column and chemiluminescence. , 1991, Analytical biochemistry.

[8]  A. Skillen,et al.  Detection of NADH via hydrogen peroxide at a platinum electrode using immobilized lactate dehydrogenase and lactate oxidase. , 1991, Biochemical Society Transactions.

[9]  G. Lubrano,et al.  Determination of lactate in human saliva with an electrochemical enzyme probe , 1991 .

[10]  R. Smolenski,et al.  Purines, lactate and phosphate release from child and adult heart during cardioplegic arrest. , 1990, Clinica chimica acta; international journal of clinical chemistry.

[11]  M. Ho,et al.  Flow injection analysis of L-lactate with enzyme amplification and amperometric detection. , 1990, Analytical chemistry.

[12]  Elo Harald Hansen,et al.  Exploitation of the Flow Injection Approach for Analytical Procedures Based on Enzymatic Amplification Reactions , 1990 .

[13]  L. Kaplan,et al.  Methods in Clinical Chemistry , 1987 .

[14]  F. Ceriotti,et al.  Erythrocytic glucose-6-phosphate dehydrogenase measured by a differential pH technique. , 1987, Clinical chemistry.

[15]  A. Mosca,et al.  Measurement of Erythrocyte Acetylcholinesterase and Plasma Cholinesterase Activity by a Differential pH Technique , 1986, Annals of clinical biochemistry.

[16]  J. Karlsson.  Muscle exercise, energy metabolism and blood lactate. , 1986, Advances in Cardiology.

[17]  M. Massi-Benedetti,et al.  An L-lactate sensor with immobilized enzyme for use in in vivo studies with an endocrine artificial pancreas. , 1985, Clinical chemistry.

[18]  J A Davis,et al.  Anaerobic threshold: review of the concept and directions for future research. , 1985, Medicine and science in sports and exercise.

[19]  Fumio Mizutani,et al.  An enzyme electrode forl-lactate with a chemically-amplified response , 1985 .

[20]  A. Mosca,et al.  Measurement of glucose in plasma by a differential pH technique. , 1983, Clinical chemistry.

[21]  H. Bergmeyer.  Methods of Enzymatic Analysis , 2019 .