Study On Extraction And Determination Of Multi-Component In Viticis Fructus And Paeoniae Radix Alba And Homogenization Of Decoction Pieces Of Farfarae Flos Based On Experimental Design

The quality control of traditional Chinese medicine?TCM?is the basis for the modernization of TCM and the key factor to ensure the safety,effectiveness and stability of TCM.TCM was defined that the medicinal herbals,animals,minerals and their preparations,which were recognized and used to treat desease under the guidance of TCM theory.However,TCM theories often lack the scientific evidences under the modern medical model,which in turn leads to some problems in methods of quality control of TCM.Taking Viticis Fructus?VF?,Paeoniae Radix Alba?PRA?and Farfarae Flos as examples,some quality evaluation methods combined with chemometrics were established based on their respective problems.It was the research objective to improve or supplement the quality control methods for TCM,and lay the foundation forfurther research of TCMs.1.A Beta/ZSM-22 zeolites-based-mixed matrix solid-phase dispersion method with HPLC-DAD was firstly developed for simultaneous extraction and quantification of eight compounds with different polarities from Viticis Fructus including two phenolic acids?protocatechuic acid and p-hydroxybenzoic acid?,two iridoid glycosides?agnuside and 10-O-vanilloylaucubin?,one benzaldehyde?vanillin?and three flavonoids?luteolin,DHTMF and casticin?.The mixed sorbent consisting of Beta and ZSM-22 was adopted for matrix solid-phase dispersion method to extract multiple compounds in VF.The three-phase mixture made of water,tetrahydrofuran and methanol was used as eluent in MSPD method.The essential parameters including types of sorbents,mass ratio of Beta to ZSM-22,mass ratio of matrix to sorbent,grinding time,types,concentration and volume of eluent were studied to obtain optimal extraction yield.Compared to the traditional MSPD method,it was easy to obtain the maximum extraction yield for simultaneous extracting hydrophilic and lipophilic constituents utilizing the mixed sorbents.The proposed mixed matrix solid-phase dispersion method was expected to be beneficial for the extraction and determination of constituents across a great polarity span.2.The persistent calyx,peel and flesh of Viticis Fructus were peeled off,and eight compounds of them were determined,respectively.To research the distribution of each constituent in different part of Viticis Fructus,the contents of target compounds were weighted by the mass ratio of different part to total.Casticin,the quality mark of Chinese Pharmacopoeia,was mainly distributed in the peel.The components in its persistent calyx have little contribution to the whole fruit of Viticis Fructus.Therefore,there is a certain material basis in“remove the persistent calyx form fruit of Viticis Fructus”.3.A green?-CD-based microwave-assisted extraction technique for the simultaneous quantification of six constituents in Paeoniae Radix Alba was established.Several important factors were optimized by the single-factor experiment and Box-Behnken design experiment.The yields of monoterpene glycosides?oxypaeoniflorin,albiflorin and paeoniflorin and benzoylpaeoniflorin?,galloyl glucoses?PGG?and phenolic compounds?gallic acid?reached the maximum level compared to another method.The kinetic and thermodynamic results demonstrated the existence of optimal extraction parameters.The proposed?-CDMAE with HPLC-DAD method was expected to replace the traditional organic solvent-based extraction method for processing sample and evaluating quality of PRA.4.To establish the HPLC fingerprints of water soluble extract of Paeoniae Radix Alba and to simultaneously determine the contents of gallic acid,oxypaeoniflorin,albiflorin,paeoniflorin,1,2,3,4,6-Penta-O-Galloyl-Beta-D-Glucopyranose and benzoylpaeoniflorin.The analysis of aqueous extract of this herbal medicine was performed on a 30?thermostatic Agilent Eclipse Plus C₁₈ column?4.6 mm×250 mm,5?m?,with the mobile phase comprising of 0.1%formic acid solution?A?-acetonitrile?B?flowing at 1.0 m L/min under gradient elution mode and the detection wavelength was set at 230 nm and 258 nm,respectively.There were nine common peaks in the fingerprints of fifteen batches of samples,five of which were identified.The similarities were more than 0.99.Good linear relationships were obtaine for six constituents within their own ranges?R²?0.9998?.All average recoveries of six constituents were 94.4%-105%with the RSDs of 1.05%-4.05%.The accurate and reproducible method can be used for the quality control of water soluble extract of Paeoniae Radix Alba.5.To optimize the homogenization method of the crude and honey-stir-baked Farfarae Flos standard decoction pieces.The content of tussilagone was determined by HPLC.The content of tussilagone,flour yield and alcohol soluble extract were selected as markers to optimize the homogenization of the crude and honey-stir-baked Farfarae Flos standard pieces.The homogenization method of the crude Farfarae Flos was A₃B₃C₃ with crushing 110 g materials 3 times,and smashing 75 s each time,while that of the honey-stir-baked was A₂B₂C₂ with crushing 60 g materials 2 times,and smashing60 s each time.The HPLC method showed good linearity?r=0.9996?for the examined concentration ranges for tussilagone,and the average recoveries of tussilagone was103%.The homogenization method is stable and feasible for the crude and honey-stir-baked Farfarae Flos standard pieces.