The prediction accuracy of visible and near-infrared (Vis-NIR) spectroscopy for soil chemical and biological parameters has been variable and the reasons for this are not completely understood. Objectives were (1) to explore the predictability of a series of chemical and biological properties for three different soil populations andbased on these heterogeneous data sets(2) to analyze possible predictive mechanisms statistically. A number of 422 samples from three arable soils in Germany (a sandy Haplic Cambisol and two silty Haplic Luvisols) of different long-term experiments were sampled, their chemical and biological properties determined and their reflectance spectra in the Vis-NIR region recorded after shock-freezing followed by freeze-drying. Cross-validation was carried out for the entire population as well as for each population from the respective sites. For the entire population, excellent prediction accuracies were found for the contents of soil organic C (SOC) and total P. The contents of total N and microbial biomass C and pH were predicted with good accuracy. However, prediction accuracy for the other properties was less: content of total S was predicted approximately quantitatively, whereas Vis-NIR spectroscopy could only differentiate between high and low values for the contents of microbial N, ergosterol, and the ratio of ergosterol to microbial biomass C. Contents of microbial biomass P and S, basal respiration, and qCO(2) could not be predicted. Prediction accuracies were greatest for the entire population and the Luvisol at Garte, followed by the Luvisol at Hohes Feld, whereas the accuracy for the sandy Cambisol was poor. The poor accuracy for the sandy Cambisol may have been due to only smaller correlations between the measured properties and the SOC content compared to the Luvisols or due to a general poor prediction performance for sandy soils. Another reason for the poor accuracy may have been the smaller range of contents in the sandy soil. Overall, the data indicated that the accuracy of predictions of soil properties depends largely on the population investigated. For the entire population, the usefulness of Vis-NIR for the number of chemical and biological soil properties was evident by markedly greater correlation coefficients (measured against Vis-NIR predicted) compared to the Pearson correlation coefficients of the measured properties against the SOC content. However, the cross-validation results are valid only for the closed population used in this study.