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Proceedings Paper

Disposable cuvette test for enzymatic determination of heavy metals
Author(s): Otto S. Wolfbeis; Claudia Preininger
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Paper Abstract

We report on an optical cuvette test for total heavy metals based on the inhibition of the enzyme urease by metals ions including silver(I), mercury(II), copper(II), nickel(II), cobalt(II), and cadmium(II). The enzymatic action is monitored using an optical ammonia transducer deposited on the wall of a disposable cuvette. This results in a rapid and inexpensive single-shot device for heavy metal sensing. A solution of urease and buffer is placed in the cuvette with the ammonium sensor membrane fixed on one of its walls. Enzymatic action starts after addition of a defined quantity of urea. This is indicated by the increase in the absorption of the ammonia sensor membrane whose color changes from yellow to blue. The slop of the increase in signal is the information for the un-inhibited reaction. After several minutes,the sample (containing the heavy metal) is added to the cuvette. Heavy metal ions inhibit the enzyme (by binding to the sulfhydryl groups) and cause a decrease in the slope. The ratio of slopes of un-inhibited and inhibited reactions is a direct parameter for detecting and calculating total heavy metals. The optimum pH was a trade-off between optimum enzyme activity (pH 7 at 25 degree(s)C) and the relative signal change of the ammonia-sensor (highest at pH 8). pH 7.5 was found to be optimal. The system was calibrated at optimized activities of urease (1.5 (mu) ) and an optimized urea concentration (0.5 mmol). Heavy metals inhibit in the following order: Ag(I) > Hg(II) > Cu(II) >> Ni(II) > Co(II) > Cd(II) > Fe(III) > Pb(II), Zn(II). The following concentrations that cause 50% inhibition were found: Ag(I) (0.1 ppm), Hg(II) (0.5 ppm), Cu(II) (0.5 ppm), Ni(II) (7 ppm), Co(II) (30 ppm), Cd(II) (95 ppm), Fe(III) (50 ppm), Zn(II) (85 ppm) and Pb(II) (210 ppm). We also studied the inhibitory effect of combinations of metal ions, the influence of ionic strength, and the effect of incubation time.

Paper Details

Date Published: 9 October 1995
PDF: 1 pages
Proc. SPIE 2504, Environmental Monitoring and Hazardous Waste Site Remediation, (9 October 1995); doi: 10.1117/12.224095
Show Author Affiliations
Otto S. Wolfbeis, Karl-Franzens-Univ. Graz (Austria)
Claudia Preininger, Karl-Franzens-Univ. Graz (Austria)


Published in SPIE Proceedings Vol. 2504:
Environmental Monitoring and Hazardous Waste Site Remediation
Tuan Vo-Dinh, Editor(s)

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