| Objective: To establish a qualitative and quantitative method for the analisis of Sanchen Pill based on the pattern that use fringerprint combined with Quantitative.To simultaneous determine seven ingredients(Syringin,Hydroxysafflor yellow A,Rutin,Kaempferol-3-Orutinoside,Anhydrosafflor yellow B,Cholic acid and Hyodeoxycholic Acid)content in Sanchen Pill by quantitative analysis of multicomponents by single marker(QAMS).The theory of serum pharmacochemistry of traditional Chinese medicine(TCM)is used to establish the serum HPLC-DAD fingerprint and study the blood constituents of Sanzchen Pill.Methods:(1)With Hydroxysafflor yellow A as the indexes,analysised of 10 batches of samples of Sanzchen Pill HPLC-DAD fringerprint in ivitro,compared with the mixed control solution to identify common peaks.(2)With Hydroxy-safflor yellow A as the indexes,calculated the relative correction factor(RCFs)between other 6 components,and which was used to calculate the conten of each component,at the same time the content of the 6components determined by the external standard method,and to compare the two methods of the calculation results,to verify the feasibility and accuracy of QAMs method.(3)Intragastriced administration(ig)to Wistar rats with 5 batches of Sanchen Pill of suspension solution,preparated drug containing serum,compared with the blank serum samples to establish Mongolian medicine Sanchen Pill serum HPLC-DAD fingerprint common model.(4)To establish a highly effective drug delivery system with different investigation of blood sampling time,frequency of intragastric administration,and method for treating serum samples.On the basis of in vivo and in vitro fingerprint of Sanchen Pill,and to compare the chromatogram of the serum samples of the whole prescription,the lack of a medicinal material group and the single herb group of Sanchen Pill,and I will analysis and explain the components which are absorbed into the blood.Results:(1)The Sanchen Pill fingerprints of10 batches were determined by HPLC-DAD method,and the evaluation model of the characteristic fingerprint of the mongolian medicine Sanchen Pill was established,and 36 common peaks were distinguished,compared with the reference sustance and confirmed that No.4 peak was syringin,No.5 peak was hydroxysafflor yellow A,No.14 peak was rutin,No.16 peak was kaempferol-3-O-rutinoside,No.18 peak was anhydrosafflor yellow B,No.24 peak was Cholic acid,No.26 peak was hyodeoxycholic acid.10 batches of samples were analyzedand the similarity were all above 0.99 with those of control samples.(2)A QAMS method determination of 7 kinds of components in the Mongolian medicine Sanchen Pill was established on the basis of the fingerprint in vitro.The content of 7 components in the 10 batches of Sanchen Pill were calculated by calibration factor and external standard method,and the results showed no significant difference.The relative correction factor was well reproduced in different types of C18 reversed phase column and different brands of chromatographic system;The 7 chromatographic peaks were determined by the method of "two point correction method" and the relative retention time.(3)The HPLC-DAD serum fingerprint of the Mongolian medicine Sanchen Pill was established successfully.5 batches of serum samples were analyzed and the similarity were all above 0.99.And identify the 6common peaks,including syringin,hydroxysafflor yellow A,kaempferol-3-O-rutinoside,anhydrosafflor yellow B,cholic acid and hyodeoxycholic acid.(4)There are 30 chromatographic peaks which have been absorbed into the blood after intragastric administration,24 of which are directly absorbed into the blood in the form of prototype,and the remaining components of the blood that may be produced by metabolism or endogenous components by the stimulation,and I will analysis and explain the components preliminary.Conclusions:(1)Established the Mongolian medicine Sanchen Pill HPLC-DAD fingerprint evaluation system,the contrast chromatogram and the 10 batchs of Sanchen Pill sample fingerprint is similar,to provide new reference for the improvement of the quality control of Sanchen pill.(2)The QAMS method can be used as a simple and accurate quality evaluation model for the quantitative determination of various components,which provides a new reference for the quality standards of the mongolian medicine Sanchen Pill.(3)Established Sanchen Pill serum HPLC-DAD fingerprint evaluation system,the contrast chromatogram and the 5 batchs of Sanchen pill in vivo fingerprint is similar,reflecting the Sanchen Pill after oral administration absorption into the blood,to provide reference for the study of Pharmacodynamic material basis.(4)Mongolian medicine Sanchen Pill has 30 constituents absorbed into the blood,the artificial bezoar contributed 3 prototype components,the remainder is contributed by safflower in direct or indirect way,the 15 components are prototype or metabolic components of safflower,9 components are provided in the compatibility of safflower and artificial bezoar,1 components are provided in the compatibility of safflower and artificial tabaschir,2 components are provided in the compatibility of three herbs.The relative peaks area are increased,include 2 components under the conditions of compatibility of safflower and artificial bezoar,as well as 4components in the compatibility of whole prescription,its showed that the 3 herbs in Sanchen Pill were play a role rogether. |
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