Add time:09/03/2019         Source:sciencedirect.com

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: 10 mM, pH 7.3 phosphate buffer, a fused-silica capillary with a total length of 49 cm × 75 μm ID, 375 μm OD with an effective length of 40 cm, 25 kV, 13 mbar 10 s pressure sample injection and 25 °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 μg mL−1 with high correlation coefficient (r = 0.9997 and 0.9993, n = 7), low limits of detection (0.47 and 0.38 μg mL−1) and quantification (1.56 and 1.28 μg 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.

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