Quality Syudy On Compound Psoralea Corylifolia L.liniment For Treating Of Vitiligo

A method for the simultaneous determination of psoralen,isopsoralen,corylin,isobavachalcone,backuchiol of Psoralea corylifolia L.by High performance liquid chromatography（HPLC）was established.Through the study of microwave extraction,accelerated solvent extraction,heating reflux extraction,ultrasonic extraction,cold extraction and other extraction methods,the extraction parameters of psoralen and isopsoralen were optimized,and the extraction process with clear parameters and controllable process was established.Based on the optimization of extraction process,using chemometrics method namely similarity evaluation,principal component analysis and cluster analysis to analysis the main component of Psoralea corylifolia L.from different origin,and to improve the consistency and quality of Psoralea corylifolia L.,in order to provide technical support for improving the quality uniformity and clinical safety.Finally,the compound of compound dexamethasone camphor liniment was made by adding Psoralea corylifolia L..Objective:Use microwave assisted method to extract the best process of coumarins in Psoralea corylifolia L.,and study the stability of extracts from Psoralea corylifolia L.by HPLC;to establish a method for simultaneous determination of psoralen and isopsoralen in Psoralea corylifolia L.by accelerated solvent extraction（ASE）and high performance liquid chromatography（HPLC）;to distinguish the medicinal materials from different sources and evaluate the quality of Chinese herbal medicines by using chromatographic profiles combined with chemometric methods;to study the method for the determination of dexamethasone acetate and camphor in compound dexamethasone acetate liniment by HPLC.METHOD:1.High performance liquid chromatography was used.The chromatographic conditions were as follows:Thermo Hypersil GOLD^TM a Q C18 column（4.6 mm×250 mm,5μm）,flow rate of 1 m L/min;column temperature:30℃;detection wavelength:246 nm,injection volume:10μL;mobile phase:methanol water（50%:50%）,the detection wavelength is 246nm.Single factor test and orthogonal test were used to optimizing the microwave extraction process of psoralen and isopsoralen.In vitro DPPH radical scavenging method was used to determine the antioxidant activity of psoralen,VC was used as the control,and the antioxidant activity was determined according to the clearance rate curve and IC50 fitting curve.2.The optimum extraction process of ASE was optimized by the contents of psoralen and isopsoralen as investigation indexes,extraction temperature,extraction time,extraction times and extraction solvent as investigation factors,and was compared with reflux extraction method and ultrasonic extraction method.3.Similarity analysis（SA）,cluster analysis（HCA）and principal component analysis（PCA）were used to analyze the original data.The chromatographic column was Waters X Select HSS T3 xp（150 mm×2.1 mm,2.5μm）,the mobile phase was acetonitrile-water（gradient elution）,the flow rate was 0.3 m L/min,the detection wavelength was 246 nm,the column temperature was 30℃,and the injection volume was 1μL.HPLC profiles of11 batches of samples were drawn by regard Psoralea corylifolia L.as the reference.Similarity evaluation software of Chinese traditional medicine chromatographic fingerprint（2012 edition）was used for similarity evaluation and analysis to determine the common peaks,and hierarchical cluster analysis and principal component analysis were conducted by SIMCA 13.0 and SPSS 22.0 software.4.The content determination was performed on column C18（4.6mm×250 mm,5μm）.The mobile phase consisted of methanol–water（70:30,V/V）.The detection wavelength was 240 nm for dexamethasone acetate,and mobile phase consisted of methanol-water（50:50,V/V）.The detection wavelength was 246 nm for Psoralen and isopsoralen,respectively.The flow rate was 1 m L/min and the column temperature was 30℃.RESULTS:1.The optimal process conditions after the investigation:0.5001g of psoralea powder（through no.2 sieve,40 mesh）was precisely weighed,the ethanol concentration was 95%,the microwave extraction power was 630w,the extraction time was 120s,and the solid-liquid ratio was 1:15.In the reaction system,psoralen has a strong scavenging ability to DPPH free radical,and shows a linear relationship.The linear approximation results show that IC50 is 3.757 mg/m L.2.The best process conditions of ASE to extraction the element of Psoralea corylifolia L.is that the extract temperature is 100℃,and the extraction time is 6 min,extraction times 3 times,50%ethanol extraction solvent,this method was used in determinate the Psoralea corylifolia L.from different regions,the results showed psoralent,and isopsoralen content range of 6.581～17.876 mg/g,respectively 6.015～16.058 mg/g.3.There were 12 common peaks in HPLC of 11 batches of psoralen samples from different regions of China.According to the similarity value of>and 0.95,11 batches of medicinal materials could be divided into 2categories.Two calculation methods of cluster analysis were adopted,and the classification results were basically consistent,indicating that the samples had good consistency.The cumulative variance contribution rate of the three principal component factors was 94.524%,and S8 sample had the highest comprehensive score and the best overall quality.4.The linear relationship of Dexamethasone was 6.2875～402.4000g/ml（R²=0.9992）,psoralen was 4.008～400.8 g/ml（R²=0.9992）.isopsoralen was 4.004～400.400 g/ml was good,and the average recovery of Dexamethasone,psoralen,isopsoralen was 108.534%（RSD%=2.42%）,84.537%（RSD%=2.73%）and 93.830%（RSD%=1.61）,respectively.Conclusion this method is simple,accurate and reproducible,suitable for the determination of dexamethasone acetate,psoralen acetate and isopsoralen in compound dexamethasone acetate liniment.Conclusion:In this paper,single factor and orthogonal experiments were used to select the best extraction process for microwave and accelerated solvent extraction of the main pharmacodynamic components in Psoralea corylifolia L..Then,ultra-high performance liquid chromatography combined with chemometrics was used to control the quality of Psoralea corylifolia L.from different places of origin in China and analyze and evaluate the main pharmacodynamic components in the drug.Finally,through investigation The quality evaluation and analysis of different medicinal adjuvants and the main effective components in liniment,the preparation of compound Psoralea corylifolia L.liniment which can improve the therapeutic effect of vitiligo,and also provides help for clinical treatment of vitiligo.