Box-Behnken响应面法优化瓜蒌皮中总核苷和总氨基酸的提取工艺

Optimization of the extraction technology for total nucleosides and total amino acids in Trichosanthis Pericarpium by Box-Behnken response surface methodology

目的采用Box-Behnken响应面法优化瓜蒌皮提取工艺,以期最大程度同时保留瓜蒌皮提取物中总核苷和总氨基酸的含量。方法采用紫外-可见分光光度法测定瓜蒌皮提取物中总核苷和总氨基酸的吸光度,并在单因素试验的基础上,以总核苷和总氨基酸吸光度为评价指标,采用Box-Behnken响应面法考察与优化料液比(g·ml^(-1))、超声提取时间(min)、醇沉浓度(%)、静置时间(h)对总核苷和总氨基酸提取量的影响。结果最佳的提取工艺为选择料液比(g·ml^(-1))为1∶15,提取时间为60min,醇沉浓度为50%,静置时间为24h。经5次平行验证试验结果表明,总核苷吸光度平均值为0.785,RSD=0.42%,模型的预测结果为0.788,实际测定结果与模型预测结果差值为0.003;总氨基酸吸光度平均值为2.112,RSD=0.07%,模型的预测结果为2.168,实际测定结果与模型预测结果差值为0.056。结论采用Box-Behnken响应面法优化的提取工艺可同时最大程度保留瓜蒌皮中总核苷和总氨基酸含量,提取工艺稳定可靠,可用于瓜蒌皮中总核苷和总氨基酸的提取。

Objective To optimize the technology for extracting Trichosanthis Pericarpium by Box-Behnken response surface methodology in order to retain the contents of total nucleosides and total amino acids in the extract of Trichosanthis Pericarpium to the greatest extent.Methods This study determined the absorbance of total nucleosides and total amino acids in Trichosanthis Pericarpium extract by UV-vis spectrophotometry.On the basis of single factor test,Box-Behnken response surface methodology was adopted to investigate and optimize the effects of solid/liquid ratio(g·ml^(-1)),ultrasonic extraction time(min),alcohol precipitation concentration(%)and standing time(h)on the extraction amount of total nucleosides and total amino acids,with the above-mentioned absorbance as the evaluation index.Results The optimum conditions for extraction were as follows:1∶15as the solid-liquid ratio(g·ml^(-1)),60min as the extraction time,50%as the alcohol precipitation concentration and 24has the standing time.The results of five parallel validation tests showed that the mean value of total nucleoside absorbance was 0.785,with RSD=0.42%.The predicted result of this model was 0.788,and the difference between the actually determined result and the predicted result was 0.003.The mean value of total amino acid absorbance was 2.112,with RSD=0.07%.The predicted result of this model was 2.168,and the difference between the actually determined result and the predicted result was 0.056.Conclusion The extraction technology optimized by Box-Behnken response surface methodology can retain the contents of total nucleosides and total amino acids in Trichosanthis Pericarpium to the greatest extent.The extraction process is stable and reliable,which can be used for the extraction of total nucleosides and total amino acids in Trichosanthis Pericarpium.