Optimization of extraction of active ingredients from toad skin and related antibacterial experiment in vitro
Objective To determine the index weight of alkaloids,cinobufagin,resibufogenin,and bufalin in toad skin,optimize the extraction of toad skin,and study the antibacterial activity of toad skin extract.Methods Based on the single-factor experiments,the ethanol concentration,solid-liquid ratio,reflux time,and reflux times,the content of alkaloids,cinobufotalin,resibufogenin,and bufalin were used as evaluation indexes,and the weight coefficients were determined with hierarchy analysis.Combined with the orthogonal experiments,the extraction parameters of toad skin were optimized,and their antibacterial activities were determined by the multiple dilution method.Results The weight coefficients for alkaloids,cinobufagin,resibufogenin,and bufalin were 0.3507,0.3507,0.1892,and 0.1093,respectively.The optimal extraction included 80％ethanol concentration,solid-liquid at 1:20,two extractions and 2.5 h each time.The extract had certain inhibition on Escherichia coli,Bacillus subtilis and Staphylococcus aureus.Conclusion The extraction process of toad skin optimized by hierarchy analysis combined with orthogonal experiment is stable,feasible and reproducible.
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