Separation And Purification Of Alkaloids From Corydalis Yanhusuo W.T. Wang And The Fingerprinting Study

This paper took alkaloids of Corydalis yahusuo W.T.Wang(C.yanhusuo) as the research mode.According to the problems need to be solved in separation and quality control of alkaloids,we developed a new process for producing alkloids with high purity using ion-exchange resins separation and high-speed counter-current chromatography(HSCCC) purification.Furthermore,the digitized fingerprints were constructed for quality control of alkaloids-containing products.The main results are listed as follows:After examined the extraction methods of C.yanhusuo,ion-exchanger resin D152 was selected for dl-tetrahydropalmatine(dl-THP) separation from crude extract of C.yanhusuo. Under the optimized sorption-desorption conditions,the area percent of dl-THP in crude extreact increased from 4.11%to 21.13%with a recovery of 65.50%using D152 packed column.Compared with liquid/liquid extraction and deposition method,ion-exchange resin showed obvious advantages of high recovery,high treatment capability,easy operation and easy regeneration.In the high-speed counter-current chromatography(HSCCC) technique, solvent screening took over 90%work time.We established a fast solvent screening method named average polarity(p').Using the new method,a HSCCC separation was further constructed to purify dl-THP with n-hexane-ethyl acetate-methanol-water(4:6:5:5,v/v) as solvent systems,and the purity of dl-THP reached 92.7%with a recovery of 95.1%after one run HSCCC purification.Fingerprint becomes a popular and authoritative method for the quality control of natureal product,which was recommended by authorities including SFDA,FDA,WHO and the European Agency.A HPLC fingerprint has been developed for the purpose of quality control of TCM product.In order to eliminate the effect of operation and reduce the fluctuation of chromatography information,a digitized fingerprint was further constructed using RRT as nature determination criterion,RPA as quantification criterion,n main peaks, difference ratio,superpose ratio and correlation coefficient as parameters,and character curve was plotted to show visual representation the similarity of different sample.This technique was expected to apply to the fingerprinting research of other natureal products.