Flow-injection assay of catalase activity.

A novel flow-injection assay (FIA) system with a double line for catalase activity was constructed in which an oxidase is immobilized and the substrate is continuously pumped to reduce the dissolved oxygen and to generate a given level of hydrogen peroxide. The catalase in a sample decomposed the hydrogen peroxide, and thus the increase in dissolved oxygen dependent on the activity was amperometrically monitored using a Clark-type oxygen electrode. Among the examined several oxidases, uricase was most suitable for the continuous formation of hydrogen peroxide from a consideration of the stability and the conversion efficiency. Under the optimum conditions, a linear calibration curve was obtained in the range from 21 to 210 units/mg and the reproducibility (CV) was better than 2% by 35 successive determinations of 210 units/ml catalase preparation. The sampling frequency was about 15 samples/h. The present FIA system was applicable to monitor the inactivation of catalase by glycation.

[Beta-lactamase activity and susceptibilities to antibiotics among some species of bacteria isolated from medical institution between December 1999 and February 2000].

We studied the beta-lactamase activity and susceptibilities to antibiotics in 604 strains among 10 species of bacteria isolated from 10 medical institutions in Tottori and Shimane Prefectures between December 1999 and February 2000. beta-Lactamase activity was measured by the nitrocefin test and penicillinase/cephalosporinase activities were measured by acidometry. beta-Lactamase activity was detected in 72.1% of S. aureus, 18.8% of H. influenzae, and 96.3% of M. catarrhalis. Penicillinase/cephalosporinase activities were detected in 17.8%/22.2% of E. coli, 9.7%/0.0% of K. pneumoniae, 18.6%/95.3% of E. cloacae, 12.7%/79.4% of S. marcescens, and 7.1%/31.8% of P. aeruginosa. Three of 72 strains (4.2%) of K. pneumoniae and 5 of 90 strains (5.6%) of E. coli were assessed as ESBL-producing bacteria using the NCCLS proposed screening method based on routine susceptibility testing results. BLNAR were detected in 13 of 69 strains (18.8%) of H. influenzae.

Flow injection analysis of DPPH radical based on electron spin resonance.

In order to construct a rapid and selective determination system of free radicals, we developed an FIA system using an electron spin resonance (ESR) spectrophotometer (flow injection spin analysis) equipped with a flow-through flat cell. In the present investigation, 1,1-diphenyl-2-picrylhydrazyl (DPPH) was used as a model free radical. Using a single line flow system, 0.5 mM DPPH was repetitively injected. When the magnetic field was fixed at 335.3 mT, the largest change in the ESR signal was observed and obtained peak height was proportional to the concentration of DPPH radical. A double line flow system was constructed in which a carrier stream containing 0.15 mM DPPH was fed into the flat cell after confluence with a sample stream. When ascorbic acid was injected as a typical DPPH radical scavenger, a negative peak appeared in proportional to the concentration. Lower detection limit of ascorbic acid was 0.01 mM (S/N=4), sampling frequency was 13 samples per h, and a satisfactory reproducibility (CV=3.2%, 0.1 mM, n=5) was obtained. The present system was also applied to estimate the DPPH radical-scavenging activity of other substances and food samples.