Studies On The Extraction, Purification And Quality Standard Of Antihypertensive Effective Parts In Huai Chrysanthemum Morifolium In Henan Province

Objective: Huai Chrysanthemum morifolium (Chrysanthemum morifolium Ramat.),the dry flower heads. As a common traditional Chinese medicine, it can clear heat,calm the liver and improve eyesight. It has been widely applicated in manyprescriptions. This study screened antihypertensive effective parts in HuaiChrysanthemum morifolium in Henan Province by observed the effects onspontaneously hypertensive model rats. The optimum extraction and purificationprocess conditions were optimized. The optimum TLC conditions were optimized.The optimum HPLC conditions were optimized. It studied on the quality standard ofantihypertensive effective parts in Huai Chrysanthemum morifolium in HenanProvince.Methods: In the experiment, the optimum extraction solvent was optimized. Theoptimum extraction conditions were optimized by orthogonal experiment. The bestextraction factors, such as concentration of ethanol, the amount of ethanol, extractiontime and extraction times were compared and determined. The optimum purificationprocess was optimized by thin-layer chromatography (TLC) and HPLC betweenmacroporous resin column and extraction method. In the study of TLC, thedeveloping solvent was optimized. In parts of the content determination, the contentof total flavonoids was determined by UV spectrophotometry and visiblespectrophotometry. The content of4-Dicaffeoylquinic acid, Chlorogenic acid, Neochlorogenic acid, Luteoloside, Isochlorogenic acid B, Isochlorogenic acid A andIsochlorogenic acid C were determined by high performance liquid chromatography(HPLC). In the pharmacology experiment, time-effect relationship was determined bymeasuring tail artery systolic pressure of spontaneously hypertensive model rats at thefirst hour, second hour, third hour, fourth hour of the first day, the third day and theseventh day after intragastric administration. Dose-effect relationship was determinedby measuring tail artery systolic pressure of high, medium and low dose(0.82g/kg.d,0.41g/kg.d,0.205g/kg.d) group spontaneously hypertensive model rats. Intragastricadministration with the optimal dose had been determined lasted for two weeks. Theimpact on spontaneously hypertensive model rats was observed by measuring tailartery systolic pressure.Results:(1) The optimum extraction solvent was ethanol. The optimum extractionprocess was:80%ethanol, twice, each time1h, the first liquid ratio was15:1, thesecond liquid ratio was10:1.(2) The optimum purification process was the extractionmethod: Took the concentrate, added in acetic acid to pH=2, added in ethyl acetate as1:1, extracted twice, combined ethyl acetate layers, added in sodium hydroxide to pH=7, recovered ethyl acetate, concentrated, and dried under reduced pressure.(3) Theresults showed that the best developing solvent was ethyl acetate-formic acid-water(8:1:1). The TLC identification methods were distinct and highly specific and couldbe used for qualitative identification.(4) The content of total flavonoids in the extractswere41.35%,48.98%measured by UV spectrophotometry and visiblespectrophotometry. The HPLC quantity determination method was simple andaccurate, with good reproducibility and recovery rates. The chosen analysis methodscould be used for the quality control of the total flavonoids in Huai Chrysanthemummorifolium in Henan Province.(5) The results showed that the best time-effectrelationship was the fourth hour of the third day after intragastric administration. Thebest dose-effect relationship was0.41g/kg.d. The total flavonoids could reduce bloodpressure of the spontaneously hypertensive model rats significantly. The differencebetween the model group and medication administration group was significant.Conclusion: The reasonable and feasible extraction and purification process were determined. Results showed that the quality control methods were simple,reproducible and accurate, and could control the quality of the extract effectively. Thetotal flavonoids in Huai Chrysanthemum morifolium in Henan Province could reduceblood pressure of the spontaneously hypertensive model rats significantly. Thepharmacological activity was clear. The experimental study provided reference forin-depth study on Huai Chrysanthemum morifolium in Henan Province in the futureand laid a good foundation for the development of safe and effective naturalantihypertensive drugs.