Studies On The Standard And Methods For Quality Control Of Wikstroemia Chamaedaphne Meisn

Wikstroemiae Chamaedaphne Meisn originates from the dried leaves and buds of thymelaceae.It is cultivated in the China provinces of HeBei,ShanXi,HeNan and other places.Wikstroemiae Chamaedaphne Meisn beginning in Shen Nong's Herbal,Bencao Tujing and Chinese Material Medica and Compendium of Materia Medica also recorded writings,contains in the “standard of Chinese medicinal materials in Shanxi Province”(1987)and “Chinese Pharmacopoeia” 1977 edition.It has the effect of purging by water and can be used in the treatment of water swelling,phlegm retention,cough,acute gasp,chronic hepatitis,schizophrenia,epilepsy and other diseases.In order to improve and revise local standards,this research has carried on character identification,microscopic and thin layer chromatography identification to Wikstroemiae Chamaedaphne Meisn,determination of the moisture,total ash content,acid insoluble ash content and the effective composition content,which provides the basis for quality control of Wikstroemiae Chamaedaphne Meisn.On the basis of the above,we create the fingerprint of 12 batches of Wikstroemiae Chamaedaphne Meisn by the method of HPLC-UV.Precision,repeatability and stability are good,which is helpful for quality control of Wikstroemiae Chamaedaphne Meisn.Daphne Genkwa originates from the dried leaves and buds of thymelaceae and is similar to Wikstroemiae Chamaedaphne Meisn and slightly larger.Wikstroemiae Chamaedaphne Meisn and Daphne Genkwa have the similar pharmacological effects and both can be used in the treatment of water swelling and phlegm retention syndrome.This study established a HPLC-MS/MS method for simultaneous determination 13 flavonoids and 3 phenolic acids of Wikstroemiae Chamaedaphne Meisn and Daphne Genkwa.This method was used to compare and evaluate the different origins of 16 Wikstroemiae Chamaedaphne Meisn and 11 Daphne Genkwa.As a result,it could provide an effective method to control the quality of Wikstroemiae Chamaedaphne Meisn and Daphne Genkwa.Part one Studies on quality standard of Wikstroemia Chamaedaphne MeisnObjective: To establish quality standards of Wikstroemia Chamaedaphne Meisn and control effectively the quality of Wikstroemia Chamaedaphne Meisn.Methods: Medica textual research through the literature on Wikstroemia Chamaedaphne Meisn,character identification,microscopic identification and thin layer chromatography identification were built for 12 batches of samples,determinations of moisture,total ash,acid insoluble ash and extracts of 12 batches of samples,simultaneously a method was established to determine the content of luteolin from Wikstroemia Chamaedaphne Meisn.Results: Wikstroemia Chamaedaphne Meisn base source was determined;the character of Wikstroemia Chamaedaphne Meisn and microscopic identification features of powders of Wikstroemia Chamaedaphne Meisn were summarized;TLC identification method built has a good specificity and durability;the results of moisture determination is 6.69%-9.61%,the average is 8.21%;the results of total ash determination is 4.75%-7.48%,the average is 6.25%;the results of acid insoluble ash were 0.38%-1.27%,the average is 0.63%;the results of alcohol soluble extract determination is 25.61%-33.76%,the average was 29.03%,the results of luteolin determination is 0.011%-0.050%,the average was 0.033%.Conclusion: The quality standard of Wikstroemia Chamaedaphne Meisn as established.The method is reliable and specific,which provides the basis for controlling effectively over the quality of Wikstroemia Chamaedaphne Meisn.Part two Studies on HPLC fingerprints of Wikstroemia Chamaedaphne MeisnObjective: To establish a fingerprint analysis method for Wikstroemia Chamaedaphne Meisn and compare the fingerprints of Wikstroemia Chamaedaphne Meisn collected from different producing areas so as to establish a sensitive and specific method for controlling the quality of Wikstroemia Chamaedaphne Meisn.Methods: 15 samples and 1 contrast medicinal herb were determined by high performance liquid chromatography-dia detection(HPLC-UV).The separation was performed on a DiamonsilTM C18 column(250 mm × 4.6 mm,5 ?m).Gradient eluted with a mixture consisting of acetonitrile(A)and water with 0.1% formic acid(B)at a flow rate of 0.8 m L/min,using a linear gradient of 0-5 min,10% A;5-30 min,10-20% A;30-45 min,20%-30% A;45-60 min,30%-65% A;60-65 min,65% A.The temperature of column was 30?.Results: The mutual mode of Wikstroemia Chamaedaphne Meisn fingerprints was set up,getting 16 common peaks,in which 9 peaks were identified,the similarity degrees of Wikstroemia Chamaedaphne Meisn in different areas were higher than 0.888.Conclusion: A simple and reliable method was established,which is useful for quality evaluation of the crud drug of Wikstroemia Chamaedaphne Meisn.Part three Comparision of compounds of Wikstroemia Chamaedaphne Meisn and Daphne Genkwa by LC-MS/MSObjective: To establish a LC-MS/MS method for simultaneous determination of 13 flavonoids and 3 phenolic acids in Wikstroemia Chamaedaphne Meisn and Daphne Genkwa and cluster analysis is applied to classify them.Methods: The multiple-reaction monitoring scanning was employed for the quantification with electrospray ion source in negative mode.The separation was performed on Agilent C18 column(150 mm×4.6 mm,5?m).The mobile phase consists of methanol-water with 0.1% formic acid with gradient elution,and the flow rate is 0.8 mL/min,the injection volume is 10 ?L.16 Wikstroemia Chamaedaphne Meisn and 11 Daphne Genkwa were analyzed and the cluster analysis is applied to classify and compare different sources of Wikstroemia Chamaedaphne Meisn and Daphne Genkwa.Results: 16 chemical compounds have good linear relationships,and precision,accuracy and stability all meet the requirements.The recovery rate is 94.2%-103.8% on average.The results show that the contents of luteolin and kaempherol in Wikstroemia Chamaedaphne Meisn and Daphne Genkwa are similar,differences of contents of the rest components are large.Conclusion: The method is simple,of high sensitivity and good specificity,which could be used for determination of 13 flavonoids and 3 phenolic acids in Wikstroemia Chamaedaphne Meisn and Daphne Genkwa.It is applicable to control quality of Wikstroemia Chamaedaphne Meisn and Daphne Genkwa.