Investigation for Purification Process,Characteristic Spectrum and Detoxification Effects on Heavy Metal Pb of Total Flavonoids in EtOAc Extraction from Tufuling(Smilax glabra)
Objective To optimize the purification process of total flavonoids in EtOAc extraction from Tufuling(Smilax gla-bra),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 spectro-photometry 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 spec-trum 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 purifi-cation 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 reference for elucidation on therapeutic material basis of detoxification of Pb by Tufuling(Smilax glabra).
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