独活挥发油低共熔溶剂辅助微波水蒸气蒸馏法提取及质量标志物(Q-Marker)研究

Study on extraction of volatile oils from Angelicae Pubescentis Radix by deep eutectic solvents assisted microwave steam distillation and its quality markers

目的建立独活Angelicae Pubescentis Radix挥发油的高效提取方法,并预测其质量标志物(quality markers,QMarker)。方法采用水蒸气蒸馏法(steam distillation,SD)、微波辅助水蒸气蒸馏法(microwave assisted steam distillation,MASD)和低共熔溶剂(deep eutectic solvents,DESs)辅助微波水蒸气蒸馏法(DESs assisted microwave steam distillation,DES-A-MSD)提取独活挥发油,并通过网络药理学和分子对接分析预测独活挥发油的Q-Marker。结果通过对不同体系DESs和提取方法的比较,发现氯化胆碱与乳酸物质的量比为1∶2的DESs体系更有利于提取独活挥发油。通过对SD法和MASD法的比较,发现DES-A-MSD法的挥发油得率最高,约为SD法的2倍。采用单因素法对其提取条件进行优化,得到最佳提取条件为氯化胆碱与乳酸物质的量比1∶2,微波功率500 W,含水量10%。采用气相色谱-质谱法对其挥发油的化学成分进行分析,并将β-瑟林烯、α-蛇床烯、红没药醇、蛇床子素和2-羟基环十五酮5种主要成分作为Q-Marker候选物进行网络药理学分析。搭建了组分-靶点网络、蛋白-蛋白相互作用网络、组分-靶点-通路网络和靶点组织分布网络,从而预测挥发油的Q-Marker可能是红没药醇和α-蛇床烯。经过分子对接分析,从理论上验证了预测的Q-Marker的生物活性。结论建立的DES-A-MSD法能够提高独活挥发油的得率,预测的Q-Marker能够用于独活挥发油的质量评价。

Objective To establish an efficient extraction method of volatile oil from Duhuo(Angelicae Pubescentis Radix,APR)and predict its quality markers(Q-Marker).Methods In this study,steam distillation(SD),microwave assisted steam distillation(MASD)and deep eutectic solvents(DESs)assisted microwave steam distillation(DES-A-MSD)were employed to extract volatile oils from APR.In addition,the Q-Marker for quality control of volatile oils from APR was predicted through network pharmacology and molecular docking analysis.Results It was found that a deep eutectic solvent system with a molar ratio of choline chloride to lactic acid of 1:2 was more advantageous for the extraction of volatile oils from APR by comparing DESs of different systems and extracting methods.In addition,the yield of volatile oils by DES-A-MSD method was the highest by comparing SD with MASD methods,and the yield of volatile oils was approximately twice of SD method.Then,the single factor method was used to optimize the extraction conditions of the volatile oils from APR.The optimal conditions were as follow:molar ratio 1:2,microwave power 500 W,and water content 10%.The gas chromatography-mass spectrometry(GC-MS)was employed to analyze the chemical components of the volatile oils from APR.Five main components,includingβ-selinene,α-selinene,α-bisabolol,osthole,2-hydroxycyclopentade-canone,were used as Q-Marker candidates for network pharmacology analysis.The component-target network,protein-protein interaction network,component-target-pathway network,and target tissue distribution network were constructed to predict the Q-Marker.It was predicted thatα-bisabol andα-selinene with high biological activity would be Q-Marker for the volatile oils of APR.Molecular docking was used to verify the biological activity of predicted Q-Marker.Conclusion The established DES-A-MSD method could improve the yield of volatile oils,and the predicted Q-Marker could be used to evaluate the quality of volatile oil.