Analysis of pyocyanin from Pseudomonas aeruginosa by adsorptive stripping voltammetry. 1996

D V Vukomanovic, and D E Zoutman, and G S Marks, and J F Brien, and G W van Loon, and K Nakatsu
Department of Pharmacology and Toxicology, Queen's University, Kingston, Ontario, Canada.

We have developed an adsorptive stripping voltammetric (AdSV) method that can be used for analysis of pyocyanin at the nanomolar level. The method employs a hanging mercury drop electrode (HMDE) as a working electrode in a three-electrode system in a 15-mL quartz cell. Preconcentration of pyocyanin in a 10-mM ammonia buffer (pH 8.0) on the HMDE is carried out by adsorption under conditions of controlled mass transfer for 60 sec, followed by scanning the electrode potential that results in reduction (cathodic scan) or oxidation (anodic scan) of the accumulated pyocyanin. For analysis of pyocyanin in samples containing a significant amount of surface-active species from a bacterial culture broth, a positive-going (anodic) potential scan must be applied after an adsorption at -0.50 V. Anodic scan of the adsorbed pyocyanin provided a well-defined oxidation peak at a potential of -0.17 V. The "anodic scan" variant of the method was tested for utility in Mueller-Hinton broth diluted from 20 to 200 times in a 10-mM ammonia buffer. Reproducibility of the method as applied to the analysis of pyocyanin produced by Pseudomonas aeruginosa (18.83 +/- 0.32 microM) in culture was demonstrated by the within-day and day-to-day coefficients of variation of 1.7% and 3.4%, respectively.

UI MeSH Term Description Entries
D010084 Oxidation-Reduction A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471). Redox,Oxidation Reduction
D011550 Pseudomonas aeruginosa A species of gram-negative, aerobic, rod-shaped bacteria commonly isolated from clinical specimens (wound, burn, and urinary tract infections). It is also found widely distributed in soil and water. P. aeruginosa is a major agent of nosocomial infection. Bacillus aeruginosus,Bacillus pyocyaneus,Bacterium aeruginosum,Bacterium pyocyaneum,Micrococcus pyocyaneus,Pseudomonas polycolor,Pseudomonas pyocyanea
D011710 Pyocyanine Antibiotic pigment produced by Pseudomonas aeruginosa. Pyocyanin
D002021 Buffers A chemical system that functions to control the levels of specific ions in solution. When the level of hydrogen ion in solution is controlled the system is called a pH buffer. Buffer
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D004566 Electrodes Electric conductors through which electric currents enter or leave a medium, whether it be an electrolytic solution, solid, molten mass, gas, or vacuum. Anode,Anode Materials,Cathode,Cathode Materials,Anode Material,Anodes,Cathode Material,Cathodes,Electrode,Material, Anode,Material, Cathode
D000327 Adsorption The adhesion of gases, liquids, or dissolved solids onto a surface. It includes adsorptive phenomena of bacteria and viruses onto surfaces as well. ABSORPTION into the substance may follow but not necessarily. Adsorptions
D000641 Ammonia A colorless alkaline gas. It is formed in the body during decomposition of organic materials during a large number of metabolically important reactions. Note that the aqueous form of ammonia is referred to as AMMONIUM HYDROXIDE.
D015203 Reproducibility of Results The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results. Reliability and Validity,Reliability of Result,Reproducibility Of Result,Reproducibility of Finding,Validity of Result,Validity of Results,Face Validity,Reliability (Epidemiology),Reliability of Results,Reproducibility of Findings,Test-Retest Reliability,Validity (Epidemiology),Finding Reproducibilities,Finding Reproducibility,Of Result, Reproducibility,Of Results, Reproducibility,Reliabilities, Test-Retest,Reliability, Test-Retest,Result Reliabilities,Result Reliability,Result Validities,Result Validity,Result, Reproducibility Of,Results, Reproducibility Of,Test Retest Reliability,Validity and Reliability,Validity, Face

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