The influence of different fertilizers (mineral/organic) on the quantity and quality of soil organic matter was monitored in long-term stationary experiments (27 years) with silage maize monoculture production on Luvisol. The main aim of this study was to investigate the relationship between easily extractable glomalin (EEG), total glomalin (TG), and parameters commonly used for the determination of soil organic matter quality, i.e., the content of humic acids (CHA), fulvic acids (CFA), and potential wettability index (PWI). A significant correlation was found between EEG content and CSOM content, humic acid content (CHA), humic acid/fulvic acid ratio (CHA/CFA), PWI, and index of aromaticity (IAR). Furthermore, the contents of EEG and TG correlated with soil organic carbon (CSOM). Periodical application of sewage sludge and cattle slurry increased the content of glomalin in soils. From the results, it is obvious that data about glomalin content can be used to study soil organic matter quality. A more sensitive method (a method that reacts more to changes in components of soil fertility) seems to be the determination of EEG rather than TG. The factors supporting use of EEG extraction in agronomic practice are mainly the substantially shorter time of analysis than TG, CHA, and CFA determination and lower chemical consumption. Furthermore, the PWI method is even suitable for studying soil organic matter quality. On the other hand, the humus quality ratio (E4/E6) does not provide relevant information about soil organic matter quality.