Study On Optimization Extraction Process And The Content Change In Decomposition And Serum Pharmacochemistry Of Danggui Shaoyaosan

Objective:Traditional Chinese medicine compound Danggui-Shaoyao-San（DSS）was studiedin this paper, by investigating its main components, screening optimization of theextraction method, To compare the contents of its main components in DSS decoctedtogether and decomposed formulas,establishing the fingerprint, to investigate the serumpharmacochemistry of DSS.So it can illuminate the blood transitional componentssource of dangguishaoyaosan decoctions initially and identify the pharmacokineticcharacteristics of the ingredients,and to a certain extent, it can explain the compoundcompatibility discipline and effective material basis of dangguishaoyao-san decoctions.Methods:①An orthogonal test was adopted in the study.The content of albiflorin,paeoniflorin, ferulic acid and alisol B-23-acetate was used as indices of appraisal.Aprocessing was carried out with multi-index comprehensive evaluation method foroptimizing the extraction condition.②To compare the contents of albiforin (Alb),paeoniflorin (Pae),ferulic acid (FA)and Alisol B23-acetate (Alisol B23-acetate) in blood group, water drug group, DSSdecocted together and decocted separately,the results were statistically analyzed.③Based on a A high performance liquid chromatography (HPLC) fingerprint,toestablish the fingerprint quality control standard of DSS extract including the commonpeak, similarity and peak area ratio,and detection of10batches of samples to ensure thequality stability of drug samples.④The established fingerprint are used to analysis the blood transitionalcomponents of rats after administrating the DSS and its decomposed formulas.Thentesting the preparation methods of serum sample and establishing the chromatographicfingerprint of blood transitional components of DSS.According to analysis thefingerprints of vivo and vitro compounds, the source of transitional components was definited in rats blood.Then comparing the fingerprints of DSS, DSS lacking of eachsingle herb,blood group,water drug group, and each single herb,which is to determinethe source of transitional components in blood and clarify the principal of compatibilityof herbs.⑤HPLC-DAD research on the transitional components in blood from0to6hours to identify the pharmacokinetic characteristics of the material in the transitionalcomponents.Results:①With the determination of main components of DSS in the orthogonalexperiment,75%alcohol,8times volume of crude drug, refluxed twice,1.5hour eachtime, By HPLCand calculate the percentage content of Alb,Pae,FA and Alisol B23-acetate were0.3254%,1.1749%,0.0249%and0.0514%.it equals: Alb3.2540mg,Pae11.7490mg, FA0.249mg and Alisol B23-acetate0.514mg per100gram contained incrude drugs of DSS.②To compare the contents of Alb,Pae,FA and Alisol B23-acetate indangguishaoyao-san decocted together,decocted separately,blood group and water druggroup, the4indices components are increased significantly on the content of totaldecoction (P <0.05).③The chromatographic fingerprint analysis method of DSS and its decomposedformulas was established by HPLC-DAD, identified the21common peaks, the use ofthe “correlation coefficient” and “the cosine method” calculation fingerprintingsimilarity,the results were more than above0.90,indicating that the establishment offingerprint technology indicator are stable and reproducible. Among the total21fingerprint peaks of the HPLC fingerprint chromatograms,fingerprint peaks ofNo.3,7,8,10,19,20,21are in respective accordance with tetramethylpyrazine,albiforin,pa-eoniflorin,ferulic acid,ligustilide,atractylenolide and alisol B23-acetate.FingerprintPeaks of No.2,4derive from the6flavor;Fingerprint peaks of No.1derives from Paeonia, Ligusticum chuanxiong and Alisma orientalis;Fingerprint peaks of No.5,6derive from Paeonia,Angelica sinensi and Ligusticum chuanxiong; Fingerprint peaks ofNo.12derives from Paeonia and Ligusticum chuanxiong;Fingerprint peaks of No.15,17derive from Angelica sinensi and Ligusticum chuanxiong;Fingerprint peaks ofNo.9,11,13derive from Paeonia; Fingerprint peaks of No.14,16derive from Ligusticumchuanxiong; Fingerprint peaks of No.18derives from Atractylodes macrocephala.15peaks of the21tota peaks derived from blood group,6peaks originate from water-druggroup.④There are16transitional components can be marked in the fingerprint of ratsafter administrating the DSS extract, while the paeoniflorin and ferulic acid aredetermined,7components of which are from the prototype compound ingredients, theother nine are metabolites and new produced material. It has13components in theserum of rat which administrated the single herb Paeonia,while6components can findout in DSS compoud.10components in Angelica sinensis,6components can find outin DSS compoud.18components in Ligusticum chuanxiong,8components can find outin DSS compoud.10components in Atractylodes macrocephala,4components can findout in DSS compoud.4components in Poria cocos,1components can find out in DSScompoud.1components in Alisma orientalis,1components can find out in DSScompoud.Other3transitional components are preliminary determined for newmetabolites.⑤Among the16transitional components of rats after administrating the DSSextract,indicating that14components has pharmacokinetic characteristics from0to6hours.Conclusion:①The optimal extraction method of DSS was:75%alcohol,8times volume ofcrude drug, refluxed twice,1.5hour each time. The optimized extraction technologywas rational and the extraction rate of the active components was high. ②The contents of its main components have significant differences between DSSdecocted together and decomposed formulas, providing a scientific basis to investigatethe compatibility of prescription.③Established the DSS fingerprinting in vitro, the method is sTab, reliable,reproducible, and can provide a quality control standards to reference for the chemicalcomposition of DSS extract,while laid the foundation for serum pharmacochemistry ofChinese medicines research.④This method of serum pharmacochemistry applied to research the material basisof dangguishaoyao-san extract.The results can determine the source of the serumtransitional compositions initially.To a certain extent it can explain the compatibilitydiscipline of DSS,and laid a foundation for further studying the material basis of DSS.⑤The serum pharmacochemistry research indicates that the directly substances ofDSS in vivo are transitional components and metabolites, but the transitionalcomponent is not an active ingredient,while having the pharmacokinetic characteristicsof the component may play the efficacy of the substance,the in-depth research of theseingredients will help to clarify the material basis of the compound effect.