| Objective:Saussurea involucrata was the featured Uygur medicine in Xinjiang,and it has the significant clinical effect.The experiment of studies for pharmacy and pharmacokinetic of Saussurea involucrata and Saussurea involucrata injection can provide the relevant theoretical data for the further development of Saussurea involucrata.Methods:1 The quality of ten batches of Saussurea involucrata were evaluated according to the check content of Saussurea involucrata in Chinese Pharmacopoeia 2010 edition,the highest content of the main components of rutin,chlorogenic acid and syringin in Saussurea involucrata was chosen as the major research medicinal material.The static adsorption experiment was done by the adsorption and elution of three main active components in Saussurea involucrata including rutin,chlorogenic acid and syringin with five kinds of macroporous resin to determine the best type of macroporous resin.The related parameters were investigated such as the maximum sample volume,sample concentration,sample flow rate,elution solvent and the volume of elution solvent,elution rate.The characteristics of main active components in Saussurea involucrata extracts penetrated through the membrane was evaluated by the dialysis bag,the cumulative released amount and cumulative released rate were calculated by the content of rutin,chlorogenic acid and syringin at different points,the time-percentage of drug cumulative released curve of Saussurea involucrata extracts was completed.The characteristics of membrane penetrated for Saussurea involucrata extracts in vitro can provide the relevant data for preparing the Saussurea involucrata mucosal drug delivery system.2 The fingerprints of Saussurea involucrata extracts and Saussurea involucrata injection were all established and the content of three main active components including rutin,chlorogenic acid and syringin were simultaneously determined by HPLC-UV.The correlationship research between Saussurea involucrata extracts and Saussurea involucrata injection was evaluated by fingerprints.This study can provide the reference for the further improvement of Saussurea involucrata injection preparation3 LC-MS/MS method was used for simultaneously detecting rutin,syringin and chlorogenic acid.The relevant method validation were finished,including specificity,standard curve and lower limit of quantification,precision and accuracy,stability,extraction recovery and matrix effects.After intraperitoneal administration of Saussurea involucrata injection in rats,the absorption law of the main active components including rutin,chlorogenic acid and syringin was researched by LC-MS/MS,the relevant pharmacokinetic parameters were also obtained.Results:1 Ten batches of Saussurea involucrata were all accorded with the relevant requirements in the Chinese Pharmacopoeia 2010 edition,the content of rutin,chlorogenic acid and syringin in Saussurea involucrata collected in hejing in 2012 were the highest one among the ten batches medicinal material,so Saussurea involucrata collected in hejing in 2012 was chosen as the major research medicinal material.The enrichment process of Saussurea involucrata extracts were as follows:X-5 macroporous resin was chosen as the suitable resin.The sample concentration was 50mg/mL,the sample volume was 3BV,the sample flow rate was 3BV/h,firstly the macroporous resin was rinsed with 3BV water,and then eluted with 6BV 20%ethanol,5BV 40%ethanol,the elution rate was 3BV/h.20%ethanol eluent and 40%ethanol eluent were collected,dried to get the Saussurea involucrata extracts.Chlorogenic acid and syringin in Saussurea involucrata extracts released relatively rapid in vitro by dialysis bag.The cumulative released rate of chlorogenic acid and syringin were 43.57%and 47.12%at 1h,4h cumulative released rate were at 88.39%and 86.04%.12h cumulative released rate were at 96.52%,98.28%.The cumulative released rate of rutin in Saussurea involucrata extracts was 30.74%for 1h,and 4h cumulative released rate was 69.07%,12h cumulative released rate were at 94.86%.The speed of permeable membrane of Saussurea involucrata extracts was slowly.2 The components in Saussurea involucrata extracts and Saussurea involucrata injection were determined.The relevant chromatograms of ten batches of Saussurea involucrata extracts and ten batches Saussurea involucrata injections were all obtained and incorporated into the“Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine "(2004A edition)which developed by the State Pharmacopoeia Commission,The common peak matching and similarity analysis were made.The similarity results of ten batches of Saussurea involucrata extracts and ten batches of Saussurea involucrata injections were all greater than 0.9.The characteristic fingerprints of Saussurea involucrata extracts and Saussurea involucrata injection were all established.By analyzing the fingerprints of Saussurea involucrata extracts and Saussurea involucrata injection,Saussurea involucrata extracts was relatively less in impurities and relatively higher in the content of the main active components.Saussurea involucrata injection was relatively lower in the content of the main active components and relatively more in impurities,Saussurea involucrata extracts can be used as the basis for the process improvement of Saussurea involucrata injection,it was possible to get the new Saussurea involucrata preparation.3 The LC-MS/MS method was used for simultaneously detecting rutin,syringin and chlorogenic acid in vivo.The specificity of detection method was good,the endogenous substances in plasma had no interference with the three component peaks and the internal standards peak.Three main active components of rutin,chlorogenic acid and syringin in the blank plasma of rat showed a good linear relationship.The linear ranges of rutin,chlorogenic acid and syringin in rat plasma were 1ng/mL?lOOOng/mL,1ng/mL?2000ng/mL,5ng/mL?1000ng/mL;The LLOQ of rutin,chlorogenic acid and syringin were:1ng/mL,1ng/mL and 5ng/mL.The correlation coefficient of standard curve regression equation were greater than 0.99.The precision and accuracy of the method were all investigated,the RSD of intra-day and inter-day precisions of low concentration QC samples were lower than 20%,the RSD of medium and high concentration of intra-day and inter-day precisions were lower than 15%.The RR of accuracy of low,medium and high concentrations QC samples were in the range of 85 to 115%.The RR of accuracy of low,medium and high concentration of the three active components in plasma were in the range of 85 to 115%for stability assessment,and it can ensure the accuracy and stability of the results.The extraction recovery and matrix effects of three main active components and internal standard were all meet the requirements.This method can be used for the content determination of rutin,chlorogenic acid and syringin in vivo.Saussurea involucrata injection was intraperitoneal administration,The Tmax of rutin,chlorogenic acid and syringin were all at 30min,and then the concentration of three active components in plasma declined slowly.The MRT of rutin and chlorogenic acid were at about 3h,eliminated more quickly.The MRT of syringin was about 8h,eliminated more slowly.By analyzing the pharmacokinetic data,we found that the main active components of Saussurea involucrata injection including rutin,chlorogenic acid and syringin were absorbed quickly in vivo,and the therapeutic effect was quick.Conclusion:1 The quality control of Saussurea involucrata extracts and Saussurea involucrata injection were evaluated by the fingerprints and the content determination of the main active components.The correlationship research between Saussurea involucrata injection and Saussurea involucrata extracts were investigated by fingerprints.The characteristics of main active components in Saussurea involucrata extracts penetrated through the membrane was evaluated by the dialysis bag method,the data can provide a theoretical basis for the preparation of Saussurea involucrata mucosal drug delivery system.2 LC-MS/MS method for simultaneously detecting rutin,chlorogenic acid and syringin in rat plasma was established.The methodology validation was finished at the same time.The results showed that the method was feasible.3 After intraperitoneal administration of Saussurea involucrata injections,the pharmacokinetics research of major active components were finished,which can provide the datas for the study of clinical medication. |
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