Measurement of Phenazines in Bacterial Cultures

Certain pseudomonads are capable of producing phenazines—pigmented, reversibly redox-active metabolites that induce a variety of physiological effects on the producing organism as well as others in their vicinity. Environmental conditions and the specific physiological state of cells can dramatically affect the absolute amounts and relative proportions of the various phenazines produced. The method detailed here—high-performance liquid chromatography coupled to detection by UV–Vis absorption—can be used to separate and quantify the amount of phenazines in a Pseudomonas culture. Simple spectrophotometric measurements of filtered culture supernatants can be used to quantify certain oxidized phenazines, such as pyocyanin, in cultures. For cases where the conditions under study are not planktonic cultures (e.g., soil or biofilms) extracting the phenazines may be a necessary first step.