土茯苓乙酸乙酯部位总黄酮纯化工艺、特征图谱及其对重金属铅解毒作用研究  

Investigation for Purification Process,Characteristic Spectrum and Detoxification Effects on Heavy Metal Pb of Total Flavonoids in EtOAc Extraction from Tufuling(Smilax glabra)

作　　者：黎华军 程双 彭财英[1] 王晓敏[1] 韦国兵[1] 舒积成[1] LI Huajun;CHENG Shuang;PENG Caiying;WANG Xiaomin;WEI Guobing;SHU Jicheng(Jiangxi University of Chinese Medicine,Nanchang 330004,Jiangxi,China)

机构地区：[1]江西中医药大学,江西南昌330004

出　　处：《中华中医药学刊》2024年第8期117-121,I0022,I0023,I0024,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81760721,82160660);江西省自然科学基金项目(20212ACB206006)。

摘　　要：目的优选土茯苓乙酸乙酯部位总黄酮的纯化条件,建立其特征图谱,并评价其对重金属铅解毒作用,以期为土茯苓解铅毒作用方面的开发和应用提供数据基础及参考。方法采用紫外分光光度法测定总黄酮的含量,并以此作为评价指标,考察6种不同色谱填料对土茯苓乙酸乙酯部位总黄酮纯化性能,筛选最佳纯化工艺;采用高效液相色谱法建立纯化后总黄酮的特征图谱;并以重金属铅致小鼠慢性毒性为动物模型,以铅含量、生化指标及组织形态学检测结果为指标,评价纯化后总黄酮解铅毒的活性。结果土茯苓乙酸乙酯部位总黄酮最佳纯化工艺为选用微孔树脂(MCI)填料,25%乙醇水溶液为上样溶剂冲洗3倍柱体积(VB),60%乙醇水溶液为洗脱溶剂洗脱3 VB,总黄酮含量78.0%。建立了总黄酮高效液相色谱(HPLC)特征图谱,指认了5个特征共有峰。与模型组对比,总黄酮组小鼠采食量增加,血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、肌酐(Cr)、血尿素氮(BUN)水平趋于正常,小鼠血液、肝脏和肾脏铅含量显著降低;病理切片结果显示,总黄酮对于铅诱导的肝、肾功能损伤具有一定的保护作用。结论土茯苓总黄酮纯化工艺具有结果稳定、操作方便、含量较高特点。总黄酮特征图谱精密度、重复性和稳定性良好。纯化后的总黄酮对重金属铅具有显著解毒作用。研究结果可为土茯苓解铅毒的药效物质基础阐明提供参考,并为研发解铅毒新药提供理论基础。Objective To optimize the purification process of total flavonoids in EtOAc extraction from Tufuling(Smilax glabra),establish the characteristic spectrum and evaluate the detoxification effects on heavy metal Pb,so as to provide a reference basis for development and implication of detoxification of Pb by Tufuling(Smilax glabra).Methods Ultraviolet-visible spectrophotometry was used to determine the contents of total flavonoids.The optimization of the purification process was investigated,including the adsorption and elution capacity of six different types of resins.HPLC was used to establish the characteristic spectrum of the purified total flavonoids.The chronic toxicity induced by Pb mice models was used to evaluate the detoxification effects on Pb according to the Pb content in tissue,biochemical indexes and morphological detection.Results The optimal purification conditions of total flavonoids were following,MCI was the best absorption resin,25%ethanol was used as the initial solvent and washed 3 VB,60%ethanol as elution solvent and washed 3 VB.Using the above purification conditions can obtain a total flavonoids content of more 78%.HPLC characteristic spectrum of the purified total flavonoids identified five common peaks.By comparing with the references,they were neoastilbin,astilbin,neoIsoastilbin,isoastilbin and engelitin.Compared with those of the model group,the results of slow toxicity test showed that the alanine aminotransferase(ALT),aspartate aminotransferase(AST),creatinine(Cr)and blood urea nitrogen(BUN)were reduced,the Pb contents in blood,liver and kidney of mice were significantly reduced.The morphological detection showed that the purified total flavonoids could alleviate the liver and kidney injures induced by Pb.Conclusions The purification process possess the quality of the stability,convenience and high purity.The characteristic spectrum has good precision,repeatability and stability.The purified total flavonoids showed good detoxification of heavy metal Pb.The above results can provide a r

关 键 词：土茯苓 总黄酮 纯化工艺 特征图谱 减毒 铅 

分 类 号：R284[医药卫生—中药学]