蟾皮有效成分提取工艺优化及其体外抑菌试验

Optimization of extraction of active ingredients from toad skin and related antibacterial experiment in vitro

目的确定蟾皮中吲哚生物碱、华蟾酥毒基、脂蟾毒配基、蟾毒灵含量的指标权重,优化蟾皮提取工艺,并对最优工艺下蟾皮提取物的抑菌活性进行研究。方法基于乙醇浓度、料液比、回流时间、回流次数单因素试验结果,以吲哚生物碱、华蟾酥毒基、脂蟾毒配基、蟾毒灵的含量为评价指标,运用层次分析法确定指标权重系数,结合正交试验结果优化蟾皮提取工艺参数,并采用倍比稀释法测定蟾皮提取物的抑菌活性。结果蟾皮提取物中指标成分吲哚生物碱、华蟾酥毒基、脂蟾毒配基、蟾毒灵含量的权重系数分别为0.3507、0.3507、0.1892、0.1093,最终以乙醇浓度80%、料液比1∶20、加热回流2.5 h、提取2次为最佳提取工艺。抑菌试验中,蟾皮提取物对大肠埃希菌、金黄色葡萄球菌、枯草芽孢杆菌具有良好的抑菌活性。结论层次分析法结合正交试验优化的蟾皮提取工艺稳定可行、重复性好。

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.