Optimized extraction process and identification of antibacterial substances from Rhubarb against aquatic pathogenic .

Response surface optimization was applied for the extraction of antibacterial substances from Rhubarb (ASR) against aquatic pathogenic . Based on the experimental results of single factors, the optimal extraction conditions were determined by Box-Behnken design combined with response surface methodology with conditions: 100% ethanol as extraction solvent, liquid/material ratio of 29 mL/g and extraction temperature at 88 °C for 148 min. The factual value of inhibition zones can reach 21.31 ± 0.95 mm and is not different from the predicted value (21.74 mm), which showed that the response surface methodology applied to the extraction optimization of antibacterial substances from Rhubarb against . is feasible. Moreover, the yield of ASR was 30.29 ± 2.27%. Five compounds, namely, aloe-emodin, rhein, emodin, chrysophanol and physcion, were identified in ASR by comparing the HPLC chromatogram of the reference mixtures and the sample. Contents of the five compounds were 0.68 ± 0.02, 0.24 ± 0.05, 0.78 ± 0.07, 6.68 ± 0.97 and 0.58 ± 0.15%, respectively. The minimal inhibitory concentration (MIC) values of ASR, aloe-emodin, rhein, emodin, chrysophanol and physcion were 0.625, 0.125, 0.015, > 1, > 1, and > 1 mg/mL, respectively, which indicated that aloe-emodin and rhein are the main antibacterial compounds of Rhubarb.