Abstract: 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 nM 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.

Abstract: A biosensor is described for use in FIA in which the recognition element, urease, was injected into the buffer solution passing through the sensor. A pH transducer was immersed in an internal flow-through chamber which was in contact with the sample solution via a semi-permeable membrane. Hydrolysis of urea, catalyzed by urease, caused an increase in pH which was recorded. The response time was ~3 min at 0.05 ml/min and the response was linear for up to 5 mM urea with a detection limit of 0.1 mM. Up to 28 mM glucose, 6.7 mM acetone, 0.2 mM citric acid or 0.6 mM sodium acetate did not interfere. Possible applications of this type of biosensor, incorporating other enzymes, are discussed.