The separation of humic substances from aqueous solution by ultrafiltration was investigated with flat Intersep Nadir membranes. Special consideration was given to the decrease of membrane permeability in the course of the ultrafiltration process. The decrease of membrane permeability was related to the increase of pH and calcium ion concentration in the solution. Transport properties were found to be strongly dependent on the hydrophilicity of the membranes. The membrane made of regenerated cellulose, the most hydrophilic of the membranes tested, displayed the highest permeability with respect to both water and humic substance containing solutions, and the lowest proneness to fouling by organic colloids. Analysis of the relationship between the properties of the solution and the course of the separation process revealed that the rise of pH from 4.6 to 7.0 reduced membrane fouling. The presence of mineral salts in the solution deteriorated the separation and transport properties of the membranes and intensified the fouling process.