Amperometric determination of L-lactate based on entrapment of lactate oxidase on a transducer surface with a semi-permeable membrane using a SIRE technology based biosensor. Application: tomato paste and baby food.
Determination of dissolved L-lactate in tomato paste and baby food samples using a SIRE-based (sensors based on injection of the recognition element) biosensor is reported. The measuring principle is based on the use of a small amount of enzyme, which is injected into an internal delivery flow system and held in direct spatial contact with the amperometric transducer by the use of a semipermeable membrane. Measurements are based upon the reversible enzymatic conversion of L-lactate to pyruvate and hydrogen peroxide by lactate oxidase. Differential measurements are performed in which the samples are measured in the presence and absence of enzyme allowing for control over matrix interferences present in crude samples. The linear range investigated for the determination of L-lactate in tomato paste and baby food was 0-0.1 mM using a lactate oxidase concentration of 22 U/mL. Samples were diluted with buffer prior to biosensor measurements. The L-lactate concentrations of the tomato paste and baby food were determined to be 1.02 +/- 0.02 mM and 2.51 +/- 0.10 mM, respectively, using the standard addition method. The repeatability for tomato paste and baby food measurements was 2.5% (RSD, n = 15) and 4.0% (RSD, n = 15) and the reproducibility was 13.0% (RSD, n = 45) and 3.0% (RSD, n = 45), respectively. The concentration of dissolved L-lactate can be used as a measure of freshness in the food industry. All biosensor measurements were compared with measurements from an established spectrophotometric assay (Boehringer Mannheim). It was found that the biosensor had good correlation with the spectrophotometric method. The biosensor gave 12% higher values for the tomato paste measurements and 2.5% higher values for the baby food measurements. However, a distinct advantage of the biosensor is that it can perform L-lactate measurements within 3 minutes, whereas the spectrophotometric assay requires a 35-minute measurement time.
Kriz
Kirstin
K
Chemel AB, Research Park IDEON, SE-223 70 Lund, Sweden.
Kraft
Linda
L
Krook
Margareta
M
Kriz
Dario
D
D015374:Biosensing Techniques; D004800:Enzymes, Immobilized; D019649:Food Industry / Q000592:standards; D005638:Fruit / Q000737:chemistry; D006801:Humans; D007223:Infant; D007225:Infant Food / Q000032:analysis*; D019344:Lactic Acid / Q000032:analysis*; D018551:Solanum lycopersicum / Q000737:chemistry*; D008567:Membranes, Artificial; D006899:Mixed Function Oxygenases; D011786:Quality Control; D015203:Reproducibility of Results; D014159:Transducers
D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D019344:Lactic Acid; D006899:Mixed Function Oxygenases; C021497:lactate 2-monooxygenase