Quality Control And Pharmacokinetic Study Of Phenolics In Artemisiae Argyi Folium

Artemisia Argyi Folium,the dried leaves of Artemisia argyi Levl.Et Vant.(mugwort),has the effects of warming to stop bleeding,dispersing cold and relieving pain,removing dampness and itching.It can be used to treat gastritis,skin-whitening,asthma and gynecological diseases.In the current “Chinese Pharmacopoeia(2015 edition)”,cineole is set as its only quality marker.However,with the continuous expansion of the application of Artemisia argyi,the phenolic components in this herb have gradually been valued.To improve the quality control method of phenolics,the quality control method of mugworts was establish through the contents determination combined with the in vitro bioactivity,and evaluates the quality of the geo-authentic medicinal material Qi'ai and mugwort samples of various habitats.Chemometric methods were employed to dig out key active ingredients of mugworts.The active ingredients were screened by serum pharmacochemistry experiment,and then the pharmacokinetic characteristics of the blood components and their tissue distribution were studied.Combined with experiment,the theoretical basis was provided for the study of effective substances,clinical application and drug development of mugwort.1.Quality evaluation of Artemisia argyi based on phenolic component-antioxidant activity: Using LC-MS/MS,5 kinds of phenolic acids and 17 flavonoid components in Artemisia argyi from various habitats,were determinated.The established method was rapid,comprehensive and accurate.The method was validated with specificity,linearity,precision and accuracy,sample recovery and stability experiments,which were proved to meet with the requirement of content determination.The method was applied in the content determination of 65 batches of mugworts.It was found that among the 22 components measured,the top five components with high content in 65 batches of Artemisia argyi were eupatilin,isochlorogenic acid A,B,C and chlorogenic acid;in addition,the content of 12 components in Qi'ai mugworts is significantly higher than others,revealing the authenticity of Qi'ai from the composition.Five kinds of anti-oxidation experiments,including DPPH?ABTS?FRAP?O2·-and ·OH,were used to evaluate the bioactivity of the samples from different habitates.Combined with GRA and PLS,the relevance between content and antioxidant activity of mugworts were discovered.Consequently,seven components were found to have greater contribution to the antioxidant effect of mugworts,including chlorogenic acid,isochlorogenic acid A,B,C,eriodictyol,eupatilin and jaceosidin.2.The pharmacokinetic study of the main active ingredients in Artemisia argyi: In the serum pharmacochemistry experiment,LC-MS/MS analysis was carried out to sceen the absorpted components in the rat plasma after orally given Artemisia Argyi Folium extract(AAFE).Ten main active components were identified,including two phenolic acids chlorogenic acid and isochlorogenic acid A,and eight flavonoids rutin,luteolin,capillarisin,5,7,3',4-tetrahydroxy-6,5'-dimethoxylfavone,hispidulin,6-methoxytricin,jaceosidin,eupatilin.Subsequently,an LC-MS/MS determination method was established for the ten active ingredients in rat plasma,and the method was validated by specificity,linearity,precision and accuracy,extraction recovery,matrix effect and stability.The method was applied to the pharmacokinetic study of the extract of Artemisia argyi.The pharmacokinetics study was carried out by orally administrated AAFE to SD rats,and it was found that these ten active ingredients were absorbed quickly,the Tmax was within 15 min,and rapidly declined within the next 2 h.In addition,the ten ingredients all showed second peak phenomenon,with seven components(chlorogenic acid,Rutin,Isochlorogenic acid A,Luteolin,Psyllium,Palmacyanin,and Isoflavin)have higher AUC and MRT,indicating that these compounds may have better pharmacological contribution in vivo.3.The tissue distribution study of the main active ingredients in Artemisia argyi: For distribution study,an LC-MS/MS method was established and validated for the determination of the ten active ingredients in the brain,lung,liver,kidney,heart,spleen,stomach,small intestine muscle and skin.SD rats were sacrificed 15 min,1h,2h,and 8h after administration of AAFE,and the tissues were havested.After processing,the content of ten components was analyzed by LC-MS/MS.It was found the ten active ingredients measured had the highest content in the stomach and small intestine,and also had a certain degree of enrichment in the skin and liver,which means mugwort may have the potential in the treatment of gastrointestinal diseases,whitening skin care products and liver protection products.