Activated sludge is a biological floc composed of microorganisms, organics, and inorganics and plays an important role in wastewater treatment. The pH of an activated sludge suspension determines the acid – base behaviour and the charge of the microorganisms in water. In the present study, the determination of acid – base behaviour of activated sludge was performed using microelectrophoresis and Potentiometric Mass Titrations (PMT). For activated sludge that is a complicated biological system, direct microelectrophoresis is not a suitable method for the determination of its ζ-potential. This method leads to a variety of isoelectric point (iep) values due to the presence of different microorganisms in activated sludge. On the contrary, the PMT method used for the first time for activated sludge can provide the overall point of zero charge (pzc) value that is 8.2. In addition, a simple methodology is proposed for the quantitative determination of the overall solid charge as a function of solution pH, which requires only (a) the titration curves of an electrolyte suspension and of the blank electrolyte solution and (b) the calculation of the H+ ions consumed for the above titration curves.