Rapid analysis of antimicrobial metabolites monoacetylphloroglucinol and 2,4-diacetylphloroglucinol using capillary zone electrophoresis.
A rapid capillary electrophoretic (CE) method was developed for the determination of phloroglucinol compounds, monoacetylphloroglucinol (MAPG) and 2,4-diacetylphloroglucinol (DAPG), in microbial supernatants of Pseudomonas fluorescens F113 over a 24-h growth cycle. Prior to electrophoretic separation, solid-phase extraction of supernatant samples on octadecylsilica for the purpose of sample cleanup is recommended. The optimum electrophoretic conditions were found to be 25 mM sodium tetraborate running buffer at pH 9.3, temperature at 25 degrees C with an applied voltage of 25 kV. The capillary was an Agilent fused-silica capillary of total length 33 cm x 50 microm inner diameter, 375 microm outer diameter, with effective length 24.5 cm. While MAPG and DAPG were monitored at selected wavelengths in the range of 214-320 nm, analysis at 214 nm was used and a CE separation time of less than 2 min was achieved. A partial method validation study was performed in accordance with European Agency for Evaluation of Medicinal Products (EMEA) guidelines. The method displayed linearity over the investigated range of 10-200 microg/mL, with limits of detection of 1.2 microg/mL for MAPG and 1.3 microg/mL for DAPG.
Guihen
Elizabeth
E
Microseparations Laboratory, Analytical and Biological Chemistry Research Facility, University College Cork, Cork, Ireland.
Glennon
Jeremy D
JD
Cullinane
Meabh
M
O'Gara
Fergal
F
D002021:Buffers; D019075:Electrophoresis, Capillary; D006863:Hydrogen-Ion Concentration; D010696:Phloroglucinol / Q000031:analogs & derivatives* / Q000032:analysis*; D011551:Pseudomonas fluorescens / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D002021:Buffers; C494385:monoacetylphloroglucinol; C026679:octadecylsilica; D012822:Silicon Dioxide; C059817:2,4-diacetylphloroglucinol; D010696:Phloroglucinol