The pH of microbial culture medium was calculated from equations of equilibrium, meterial balances for ionic components and electro-neutrality theory. Ammonium ion consumption and Acetic acid production are found out to be the major contributors for the alteration of the pH as well as the buffer capacity of the medium. By measuring the buffer capacity on-line, levels of acetic acid were estimated by a software sensor using pH signal in a fermentation process of E.coli growing in a minimal medium. The measured values of acetic acid showed good correlation to those of estimated by the software sensor.