| Clematidis Radix et Rhizoma was derived from the dried roots and rhizomes of Clematis chinensis Osbeck,Clematis hexapctala Pall,Clematis manshurica Rupr.It was used for the treatment of rheumatism numbness,numbness of the limbs,tension of muscles and veins,flexion and extension in clinical.According to the previous chemical studies,it was found that the chemical components of Clematidis Radix were complex,but the active components were still unknown.Therefore,It was of great significance for the scientific development and quality control of Clematidis Radix by studying and explaining the pharmacodynamic material substance basis of Clematidis Radix.Based on the clinical characteristics of topical administration of Clematidis Radix decoction,this study carried out basic research on the chemical substances of water-soluble components of Clematidis Radix.On this basis,transdermal model in vitro was used to study the transdermal behavior of its water-soluble components,and the active components were preliminarily screened.It aimed to make a valuable exploration for further research to reveal the medicinal substance basis of Clematidis Radix.In this paper,the HPLC fingerprint of Clematidis Radix was firstly studied.Through the establishment of HPLC fingerprint analysis method based on water extraction method,which was used to understand the chemical composition of the water extract of Clematidis Radix and provided analytical method support for the subsequent basic material research.The optimal ultrasonic water extraction process was obtained by comparing ultrasonic and reflux extraction methods and single factor and orthogonal test.The comparison fingerprint was established by Similarity Evaluation System of Chromatogram Fingerprint of TCM,which was based on the HPLC chromatogram of 15 batches of Clematidis Radix from different provenance and market source,and a total of 16 fingerprint peaks were identified.The results showed that the whole chemical composition of the water extract from different origins and market sources of Clematidis Radix was similar and the characteristics of HPLC fingerprints was obvious of Clematidis Radix,which could be used to distinguish the adulterants.On this basis,the paper studied the material basis of HPLC fingerprint.The chemical composition of fingerprint was studied qualitatively and quantitatively by UPLC-Q-TOF and HPLC.A total of 12 components of 8 labeled spectral peaks in the fingerprint,including 8 phenolic acids and 4 triterpenoid saponins,were identified by reference data and standard comparison.Among them,3 isochlorogenic acid compounds were reported for the first time in Clematidis Radix.And the HPLC method for simultaneous determination of 8 phenolic acids was established and validated.The results showed that caffeic acid and compound HX were the common components of all batches of medicinal materials and the content of total phenolic acid in pharmacies was significantly lower than that in market.Finally,the screening study on the active ingredients of Clematidis Radix was carried out by the transdermal characteristics.The transdermal study in vitro and HPLC fingerprint analysis were used to investigate the transdermal behavior of the compounds and it was found that the main transdermal component was compound HX by optimizing the model parameters and experimental conditions.On this basis,the influence of solvent concentration and p H value on its transdermal behavior was investigated.The results showed that the abdominal skin of defatted mice was treated with 10% ethanol-normal saline as the receiving solution.After 8 h of transdermal time,the main transdermal component of the aqueous extract of Clematidis Radix was compound HX,and its transdermal amount was positively correlated with the concentration,but negatively correlated with the p H value.The results of this paper provide reliable scientific data for further research to reveal the pharmacodynamic material basis of Clematidis Radix and the research results have certain scientific value. |
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