Soil organic matter (SOM) undergoes short and long-term transformation in the soil. Microorganisms through their enzymes are able to mineralize organic carbon while the rate of this process is different. Biological test though referred to one of the main diagnostic methods for evaluating soil quality/health. The aim of our work was to determine basal respiration, total carbon content, fractional composition of humus and basic parameters of soil colloidal complex in selected subtypes of chernozem in South Moravia region. Basal respiration was measured using Vaisala GMT220 apparatus. Total carbon content was determined by oxidimetric titration and basic parameters of soil colloidal according to Mehlich. Results showed that production of carbon dioxide varied from 0.09 to 0.27 mg CO2/100g/h. Linear correlation between basal respiration and humification degree was found. Humus content varied from 2.15% to 4.6%. No correlation between quantity of humus and basal respiration was observed. Higher values of basal respiration were connected with higher quality of HS. Significant linear correlation between total carbon content (TOC) and cation exchange capacity (CEC) was found.