Abstract：Brevibacillus brevis FJAT-0809-GLX is a biocontrol agent with broad-spectrum antifungal activities. Ethylparaben has been confirmed to be one of its main functional components, but isolation and purification of this component from the fermentation broth is very complicated and difficult. Thus, it is urgent to develope a rapid, simple and effective method for quantitative determination of ethylparabens in the fermentation broth of B. brevis FJAT-0809-GLX. An automated sysmtem using high performance liquid chromatography (HPLC) with ultraviolet detection was developed for the determination of ethylparaben in fermentation broth in the present study. The mobile phase consisted of methanol/distilled water (65∶35 V/V) using a gradient program at a flow rate of 1.0 mL/min with 254 nm detection and an injection volume of 10 μL. Good linearity was obtained over the concentration range of 1.0~20 μg/mL with the coefficient of determination of 0.998 7. For the ethylparaben, the limit of HPLC detection was 1.0×10-15 μg/mL. Subsequently, the ethylparabens in fermentation broth were detected at different time points. The results indicated that the peak times and peak areas of ethylparabens were all stable within 48 h. The average recovery rate of the additive ethylparabens from fermentation broth was 96.2%~105.3% and the relative standard deviation (RSD) was 0.8%. Finally, the ethylparaben concentration in an optimizated fermentation culture medium was detected using this HPLC determination system. The results showed that its retention time was at 3.12 min and the peak area was 136. Therefore, its concentration reached approximately 2.111 μg/mL according to the standard curve. In conclusion, a simple, sensitive and quick HPLC method was established for determination of ethylparabens in fermentation broth of B. brevis FJAT-0809-GLX. The results will lay a foundation for tracking detection of functional components from fermentation broth of B. brevis.
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