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In this study, a simple, rapid and efficient sample pretreatment technique for the determination of surfactin in microbial fermentation broths of bacterium bacillius subtitus sp., has been developed using salt-assisted homogeneous liquid-liquid extraction (SHLLE) coupled with high performance liquid chromatography/ ultra-violet detection (HPLC/UV).Surfactin has the potential to replace synthetic chemical surfactant in the near future because of its high surface activity, biodegradability and environmental-friendly nature.In order to characterizing the amount of surfactin for microbial monitoring under the fermentation production process, we have demonstrated a rapid method to analyze surfactin using a home-made extraction device.