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Abstract
A quick and sensitive high-performance liquid chromatography (HPLC) method has been developed and validated for the determination of lomofungin in Streptomyces lomondensis. Extraction of lomofungin was carried out by liquid–liquid extraction. Chromatographic separation was achieved on a C18 column (5 µm, 4.6 mm × 250 mm, Agilent Inc). The mobile phase was formic acid [0.1% (v/v)]/acetonitrile of a segmented gradient, which was 8:2 in the first 4 min, then 6:4 for 16 min, and 8:2 for 10 min. The flow rate was 1 mL min−1, and the detection wavelength was 270 nm. Under these chromatographic conditions, lomofungin was eluted at 17.6 min. The calibration curve was linear over a concentration range from 10 to 400 µg mL−1, and the accuracy was within a range of 96.6 to 104.0%. The HPLC method was applied successfully to monitor dynamic lomofungin production of Streptomyces lomondensis during a 120 hr growth cycle.
ACKNOWLEDGMENT
This research was supported by 973 Programs of China (2009CB118906 and 2012CB721005) and Shanghai Leading Academic Discipline Project, Project Number: B203.