Soil quality determination requires the analysis of a number of soil attributes using different approaches. In recent years, one of the most promising approaches has been the determination of enzymatic activities. Generally, only a few enzymes have been analysed and related to other soil properties such as total carbon, nitrogen content or microbial biomass carbon. The aim of this work was to investigate the possible use of the API ZYM strip, a semi-quantitative miniaturized system that determines 19 enzymatic activities, to study soil quality. To this end, we tested the system in different soil types, including albic Arenosols, mollic Leptosols, rendzic Leptosols, haplic Leptosols and calcaric Regosols. Fresh samples were sieved through a 2-mm sieve in the field and soil extracts were prepared by mixing 2–20 g (depending on the soil horizon characteristics) from each sample with 2–20 ml of sterile water. Next, 65-µl aliquots of the supernatant extract were placed into each API ZYM microtube and were incubated at 37°C for 16 hours. Our results show important qualitative and quantitative differences among the different soil types studied, with soil characteristics and biological properties correlating with biochemical information. The results provide useful information not only to determine soil quality, but also to assess changes in the soil environment. As a whole, our results suggest that the use of the API ZYM system could prove most useful in soil environmental studies. Finally, some suggestions are presented, including modifications to the system that could improve its application in this field.