Microplate-compatible biamperometry array for parallel 48-channel amperometric or coulometric measurements.

We report a new reusable electrochemical array for parallel biamperometric measurements that has been designed for use with standard microplates. The 48-channel array uses half of the available 96 wells and has 48 pairs of Pt wire electrodes. Applications to the quantitation of a variety of oxidizable species, including acetaminophen, ascorbic acid, hydroquinone, trolox, and uric acid, are demonstrated in assays that use potassium ferricyanide as an oxidant to produce a mixture of ferri- and ferrocyanide. Hydrogen peroxide quantitation is also demonstrated, based on an assay in which ferrocyanide is oxidized, again to produce a mixture of ferri- and ferrocyanide. Detection limits (signal-to-noise ratio (S/N) = 3) in these assays range from 1 (acetaminophen, R2 = 0.994) to 8 microM (ascorbic acid, R2 = 0.967), and linearity was observed to analyte concentrations of at least 100 microM. We also demonstrate the application of the biamperometric array to enzymatic assays, using the glucose oxidase reaction as an example; following a 20 min enzyme reaction time, a detection limit of 0.1 mM glucose was obtained. These results indicate that applications to other oxidase-based assays are feasible in this high-throughput format. The new electrochemical array employs standard, inexpensive microplates, and the biamperometric measurements are simple, precise, and rapid, requiring only 2 min for 48 parallel measurements.

Antibiotic susceptibility testing at a screen-printed carbon electrode array.

Screen-printed carbon electrode arrays were treated to allow respiratory activity-based measurement of antibiotic susceptibility with Escherichia coli JM105. Carbon working electrodes were examined for antibiotic adsorption and were pretreated with various electrochemical and chemical protocols to minimize antibiotic adsorption. Treatment by voltammetry in basic solution or by chemical modification with poly-L-lysine or chitosan were found to be effective methods for the elimination of adsorption of the studied group of 17 antibiotics, which comprised several classes and modes of action. Measurements consisted of two-electrode amperometry of the bacterial suspension after 10 min of incubation with antibiotic followed by addition of an oxidative cocktail of ferricyanide and dichlorophenolindophenol for a further 10 min; response currents, which indicate the extent of reduction of ferricyanide to ferrocyanide by cellular respiratory activity, decrease with increasing concentration of antibiotic present in the initial 10 min incubation. IC50 values obtained for chloramphenicol with these electrode modification methods are consistent at 2.0 +/- 0.2 mM, in approximate agreement with previously reported respiration-based results for this organism but significantly higher than values reported for growth-based antibiotic susceptibility testing methods.