Extraction And Purification Of The Polyphenols In Mallotus Oblongifolius (Miq.) Muell.-arg And Studied The Effect For Osteoporosis Prevention And Treatment In Ovariectomized Rats

Objective: This paper aimed to study the extraction and purification and qualitycontrol method of the polyphenols extracted from Mallotus oblongifolius (miq.)muell.-arg. To study the effect and mechanism of the polyphenols for osteoporosisprevention and treatment in ovariectomized rats.To establish the fingerprintchromatogram standard of Mallotus oblongifolius (miq.) muell.-arg by HPLC, and tostudy the pattern recognition of the fingerprint chromatogram.Method:To establish the best extraction condition of the polyphenols through singlefactor and orthogonal experiments with the the extraction rate as it’s evaluation index.To select the optimal macroporous adsorptive resins by static experiment fromAB-8,NKA-9,DM-130,HPD-600and HPD-826, and to confirm the optimaladsorption-desorption condition by dynamic experiment.Quality control of the polyphenols. To establish a TLC identification method ofpolyphenols.To analyse the content of gallic acid and EC by multi-componentsquantitation by one marker.We explored the effect and mechanism of the polyphenols for osteows.Crush themedical material into raw powder and put the powder though a24mesh sieve.Add15fold of40%ethanol,extract3times,45min per time. After filtrating,weconcentrated the filtrate to a relative density of1.1(60℃).The optimum spray dryingconditions were the inlet temperature was105℃, and outlet temperature was95℃,the pump rotatioporosis prevention and treatment in ovariectomized rats. The rats were divided randomly into6groups: the sham operation group, model group, thepolyphenols of low,medium and high dose,and positive control group. All the ratswere ovariectomized except the the sham operation group.All rats wereadministrated drugs four weeks later.After eight weeks’ administration,the rats wereanaethetized by10%chloral hydrate. The blood from the abdominal aorta was testedby E2,SOD,MDA,ALP,and we also tested index of uterus, thymus index, spleen andthe BMD and so on.We established a sensitive HPLC fingerprint method,and study the patternrecognition of the fingerprint chromatogram. HPLC analysis was performed oncolumn Dionex Acclaim120C18（250mm×4.6mm，5μm）,with a mixture ofacetonitrile and0.03%trifluoroacetic acid as mobile phase in gradient elution. Thewave-length was280nm,the floe rate was1.0ml/min, the temperature was30℃,withthe running time of70min.Results: We determined the extraction and purification condition as follon speed was7r/min, fan speed was1900r/min.We selected AB-8as the optimal macroporous adsorption resin though staticexperiment. The purification condition were as follows.The sample loading was12mg/ml,sample flow rate was1BV/h,the PH was3.Then we eluted it with waterfirst.Then we eluted it with4BV’s80%ethanol. The elution flow rate was2BV/h.Weconcentrated the ethanol elution to a relative density of1.0(60℃).We get theproduction though frozen drying.We established quality control standards of the polyphenols:We identified6components by TLC. They were catechin,EGCG,ECG,EGC,D.L-EC and the gallicacid.The TLC figure was distinct and reproducible.To analyse the content of gallicacid and EC by multi-components quantitation by one marker. This method wasaccurate and reproducible. The polyphenols can be used for osteoporosis prevention and treatment inovariectomized rats.They can increase SOD activity and the content of BMD and E2,and decrease the content of MDAandALP. They can also increase the index ofuterus.The HPLC fingerprint of Mallotus oblongifolius (miq.) muell.-arg showed15common peaks.And six components were identified,they were gallic acid, EGC,catechin, EC, EGCG and ECG. Mallotus oblongifolius (miq.) muell.-arg can bedivided into three groups according to cluster analysis and principal componentanalysis.Conclusion:The results show that the methods of extraction and purification wereSimple and feasible,and the quality was stable and reliable.And the polyphenols canbe used for osteoporosis prevention and treatment in ovariectomize rats;Weestablished the HPLC fingerprints and the pattern recognition.This paper providedscientific evidence for the development of this folk medicinal resources.