Study On Quality Control Method Of Traditional Marine Chinese Medicine Ipomoea Pes-caprae

Objective: To study on quality control method of traditional marine Chinese medicine Ipomoea pes-caprae for establishing quality of medicinal materials standard of Ipomoea pes-caprae.Methods: Regarding caffeic acid as reference substance,studied the influences of extracting solvent,chromogenic reagent,developer,temperature,humidity on TLC chromatograms,and established the TLC identification method.According to the fourth part of Chinese pharmacopoeia(2015),the content of moisture,total ash and extracts of Ipomoea pes-caprae was determined.Regarding the total content of six active components as examining index,optimized extraction technology of Ipomoea pes-caprae by response surface method.The content of six indexes in Ipomoea pes-caprae from different areas or collection time was determined by HPLC.According to traditional Chinese medicine chromatographic fingerprint similarity evaluation system(V2.0)and SPSS 19.0,HPLC and UPLC fingerprint of Ipomoea pes-caprae were established respectively.Results:(1)The TLC identification method for Ipomoea Pes-Caprae(L.)Sweet was highly specific with caffeic acid as the reference substance,cyclohexane-acetone-ethyl acetate-formic acid(6:2:1:0.15)as the developing system,1% potassium ferricyanide-2% ferric chloride as chromogenic reagent,25? as temperature,as well as 70% as humidity.(2)The critical content of moisture,total ash and extracts in Ipomeapes-caprae was temporarily confirmed as follows: the moisture content was no more than 12%,the total ash content was no more than 16%,the content of water soluble extract by hot dipping method was no less than 19%,the content of water soluble extract by cold-maceration method was no less than14%,the content of alcohol soluble extract from hot dipping method was no less than 20%,the content of alcohol soluble extract from cold-maceration method was no less than 15%.(3)The best extraction process was as follows: ethanol concentration is69%,ethanol volume is 63 m L,extracting time is 1.5 h,extracting times is 3,water bath reflux temperature is 90?.(4)The content range of caffeic acid,chlorogenic acid,isoquercitrin,isochlorogenic acid B,isochlorogenic acid A,and isochlorogenic acid C in Ipomea pes-caprae were as follows: 0.7604?12.7981 m?/?,0.1117?0.7826m?/?,0.1906 ?1.2785 m?/?,0.9892?7.6556 m?/?,2.6974?17.2133 m?/?,1.9225?9.5335 m?/?.(5)The HPLC and UPLC fingerprint were established.There are six major peaks were identified.There are 13 common peaks of HPLC fingerprint,the similarity range between ten bach samples and common pattern was 0.956 ? 1.000,Ipomea pes-caprae from different places were divided into two categories.There are 16 common peaks of UPLC fingerprint,the similarity range between ten bach samples and common pattern was 0.945? 1.000,Ipomea pes-caprae from different places were divided into three categories.Conclusion: In this thesis,the key techniques of quality control of Ipomea pes-caprae,including extraction process,qualitative and quantitative analysis,HPLC and UPLC fingerprint,routine examination items,were studied by response surface optimization techniques andchromatographic technique,as well as the chemometrics methods such as similarity analysis,cluster analysis and principal component analysis.In conclusion,all the achievement herein provided scientific methods and experimental basis for the quality control of Ipomea pes-caprae.