Study On The Quality Control Of Ilex Kudingcha And Ilex Latifolia

Ilex latifolia and Ilex Kudingcha's dry leaves are considered main source of traditional Chinese medicine Kudingcha.The current market quality of Kudingcha varies,the origin is unknown,shoddy phenomenon still exists,It is imperative to establish a quality control standard that can really react the quality of Kudingcha which has specificity.In this paper,we will study the quality control of Ilex Kudingcha and Ilex latifolia from the following three aspects: Determination of Polyphenols and Total Saponins in Ilex Kudingcha and Ilex latifolia;Determination of Monomeric Compounds of Ilex Kudingcha and Ilex latifolia;HPLC-DAD-ELSD fingerprint of Ilex Kudingcha and Ilex latifolia.The main contents of this paper are as follows:1.Analysis of Total Polyphenols of Ilex latifolia and Ilex KudingchaBy comparing the different extraction methods and solvents,the extraction method of polyphenols from Ilex latifolia and Ilex Kudingcha was determined:8 times amount of 70% ethanol,ultrasonic extraction,3 times.The polyphenols in 70% ethanol ultrasonic extraction of Ilex latifolia and Ilex Kudingcha were purified by AB-8 macroporous resin,collected water part,water bath to go to most of the solvent and vacuum drying.Using FC color method,taking chlorogenic acid as the reference substance,conduct a methodological study.The results show that the linearity,reproducibility,stability and recovery of this method are good.The content of total polyphenols in Ilex Kudingcha from different origin was range from 1.26% to 1.57%,in Ilex latifolia Thumb.from different origin was range from 0.82% to 0.87%.2.Analysis of Total Saponins of Ilex latifolia and Ilex KudingchaUsing the combination of aluminum oxide and XAD-2 macroporous resin to purify total saponins from 70% ethanol ultrasonic extraction of Ilex latifolia and Ilex Kudingcha.Collected 75% ethanol part,using cdorimetry of vanillin-glacial acetic acid,taking kudinoside D as reference substance,methodological study and the determination of the total saponins in Ilex Kudingcha were studied;taking kudinoside G as reference substance,methodological study and the determination of the total saponins in Ilex latifolia were studied.The results show the content of total saponins in Ilex Kudingcha from different origin range from 10.83% to 11.93%,in Ilex latifolia from different origin range from 7.14% to 8.10%.3.Analysis of six kinds of compounds in Ilex Kudingcha and Ilex latifoliaUsing quantitative analysis of multi-components by single marker and HPLC,taking kudinoside C as internal standard,to determine relative correction factors of Kaempferol-3-O-?-D-rutanoside,Isorhamnetin-3-O-?-D-rutanoside,Kudinoside A,Kudinoside E,Kudinoside D in Ilex Kudingcha,and calculate their contents.Investigate the reproducibility of the relative correction factor and chromatographic peak localization of the five components of different columns.At the same time,the contents of these six components were determined by external standard method(ESM),and compare the difference between ESM and QAMS to verify the feasibility and accuracy of QAMS method.Results show that the relative correction factor and chromatographic peak localization in different columns are good and there was no significant difference of the six components' contents calculated between QAMS method and ESM method.It is convenient,feasible and accurate to measure the content of six components in Ilex kudingcha by QASM method.Using HPLC to determine the content of Latifoloside F,Latifoloside H,Latifoloside C,Latifoloside D,Kudinoside F and Ilekudinoside E in Ilex latifolia,conduct a methodological study.The results showed that the linearity,reproducibility,stability and recovery of this method are good.In different origins and batches of Ilex latifolia the content of Latifoloside F range from 0.33% to 0.47%,Latifoloside H range from 0.23% to 0.32%,Latifoloside C range from 2.00% to 2.44%,Latifoloside D range from 0.32% to 0.44%,Kudinoside F range from 0.18% to 0.29% and Ilekudinoside E range from 0.15% to 0.21%.4.Study on HPLC-DAD-ELSD Fingerprint of Ilex latifolia and Ilex KudingchaThe HPLC-DAD-ELSD fingerprints of Ilex Kudingcha were established,in 10 batches Ilex Kudingcha's HPLC-DAD fingerprints,eighteen common peaks were considered as fingerprint peaks,identified seven of them,they are chlorogenic acid,Kaempferol-3-O-?-D-rutanoside,Isorhamnetin-3-O-?-D-rutanoside,Kudinoside C,Kudinoside A,Kudinoside E,Kudinoside D.The similarity of 10 batches of herbs was evaluated,range from 0.911 to 0.970.In 10 batches Ilex Kudingcha's HPLC-ELSD fingerprints,eleven common peaks were considered as fingerprint peaks,the similarity of 10 batches of herbs was evaluated,range from 0.914 to 0.962.The HPLC-DAD-ELSD fingerprints of Ilex latifolia were established,in 10 batches Ilex latifolia's HPLC-DAD fingerprints,fifteen common peaks were considered as fingerprint peaks,identified seven of them,they are chlorogenic acid,Latifoloside F,Latifoloside H?Latifoloside C,Latifoloside D,Kudinoside F and Ilekudinoside E.The similarity of 10 batches of herbs was evaluated,range from 0.915 to 0.984.In 10 batches Ilex latifolia's HPLC-ELSD fingerprints,fourteen common peaks were considered as fingerprint peaks,the similarity of 10 batches of herbs was evaluated,range from 0.909 to 0.9670.