title: Rapid quantitative analysis of phenazine-1-carboxylic acid and 2-hydroxyphenazine from fermentation culture of Pseudomonas chlororaphis GP72 by capillary zone electrophoresis.
abstract: Natural phenazines in secondary metabolites of bacteria have been receiving increasing attention in recent years due to their potential usage as antibiotics. In the present study, a rapid and reliable capillary zone electrophoresis (CZE) method was developed and validated for monitoring for the first time dynamic phenazine-1-carboxylic acid (PCA) and the 2-hydroxyphenazine (2-OH-PHZ) production of Pseudomonas chlororaphis GP72 during the entire fermentation cycle. The paper begins with the optimization of separate conditions for 2-OH-PHZ and PCA together with phenazine (PHZ), which is used as internal standard. The optimized conditions are: 10mM, pH 7.3 phosphate buffer, a fused-silica capillary with a total length of 49 cm x 75 microm ID, 375 microm OD with an effective length of 40 cm, 25 kV, 13 mbar 10s pressure sample injection and 25 degrees C air-cooling. The three compounds could be separated within 2 min under optimized conditions. The validation of the newly developed study shows the linear response of 2-OH-PHZ and PCA ranging from 10 to 250 microg mL(-1) with high correlation coefficient (r=0.9997 and 0.9993, n=7), low limits of detection (0.47 and 0.38 microg mL(-1)) and quantification (1.56 and 1.28 microg mL(-1)), respectively. Good precision values for intra- and inter-day detection and acceptable individual recovery ranges for 2-OH-PHZ and PCA are indicated. The newly developed method was also validated through monitoring dynamic PCA and 2-OH-PHZ production of P. chlororaphis GP72 during an 84 h growth cycle.
authors.affiliation: Key Laboratory of Microbial Metabolism, Ministry of Education and College of Life Science and Biotechnology, Shanghai Jiaotong University, Dongchuan Road 800, Minhang District, Shanghai 200240, PR China.
authors.forename: Hai-Ming
authors.initials: HM
authors.lastname: Liu
authors.forename: Xue-Hong
authors.initials: XH
authors.lastname: Zhang
authors.forename: Xian-Qing
authors.initials: XQ
authors.lastname: Huang
authors.forename: Cheng-Xi
authors.initials: CX
authors.lastname: Cao
authors.forename: Yu-Quan
authors.initials: YQ
authors.lastname: Xu
mesh_terms: D003470:Culture Media / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D005285:Fermentation; D008722:Methods; D010619:Phenazines / Q000032:analysis; D011549:Pseudomonas / Q000378:metabolism*; D015203:Reproducibility of Results
chemical_list: D003470:Culture Media; D010619:Phenazines; C037165:1-phenazinecarboxylic acid; C088013:2-hydroxyphenazine