Metal toxicity assessment in soils using enzymatic activity: Can water be used as a surrogate buffer?

Ecotoxicological tests based on soil enzyme activity are widely used to assess the terrestrial ecotoxicology of metals in soils. However, several standard enzymatic methods use buffers that may alter the chemical pseudoequilibrium of soils and affect metal speciation, and, in turn, the metal effects on enzymes and enzyme kinetics. Researchers have suggested the use of H₂O as a solvent rather than chemical buffers, but opponents are concerned about pH fluctuations during incubation and the resulting difficulty in comparing enzymatic studies. Enzyme assays were conducted on 10 pairs of Zn-contaminated soils to evaluate 1) the buffer effect on Zn lability 2) the pH fluctuation during enzymatic assays conducted in water and 3) the comparison of enzymatic results obtained using chemical buffers versus water. Four standard enzymatic methods covering the major biogeochemical cycles were targeted: arylsulfatase (acetate pH 5.8), urease (borate pH 10), acid phosphatase (modified universal buffer pH 6.5) and protease (THAM pH 8.1). Furthermore, deionized water was tested in parallel as a surrogate solvent for these four methods. With the exception of the acetate buffer, the tested solvents did not significantly change the labile Zn concentration in the soil samples. The pH slightly fluctuated by +0.57 pH unit, corresponding to the intrinsic variability of soils. Enzymatic methods using buffers showed similar results compared to those using water, except for urease. These observations suggest that enzymatic methods setting alkaline conditions should be used with caution and that H₂O could be used as surrogate solvent in this context.