| Objective To preliminarily study the substance of Rehmannia glutinosa Libosch on the treatment of hemostatic effect. To establish HPLC fingerprints of the hemostatic Fraction of Rehmannia glutinosa Libosch. To optimize the extraction process of Rehmannia glutinosa Libosch. to filter out the best process parameters. To select the best preparation technology of Rehmannia glutinosa Libosch Foam-rubber. To establish the quality standard of Rehmannia glutinosa Libosch Foam-rubber.Method The experimental drugs were systematically extracted by solutions (petroleum ether,ethyl acetate,n-Butanol,ethanol) from Rehmannia glutinosa Libosch according to solutions , polarity. Breaking tail hemostasia and capillary coagulation in mice were used to evaluate the hemostatic effect. The effective composition was carried out by elution method. Elutropic materials were used to do the same pharmacody-namic experiment again. Akasil C18(250 mm×4.6 mm,5μm) column was used and the acetonitrile-water was chosen as the mobile phase in a gradient mode. The column temperature was 30℃and the detection wavelength was 210 nm. The detection time was 65 min, and the flow rate was 1.0 mL /min. Taking the content of polysaccharides and the summation of the fingerprints peaks relative to the content of catalpol as the indexes.Soaking times,taking the water quantity, and times as the investigate factors to arrang the experience ,and optimize the extraction process. Using the single factor test method,take the appearance quality,water-absorbing value as an indicator to study several excipients commonly,the proportion of drug and gelatin of the two factors on the stability of the Foam-rubber Impact. TLC were used to distinguish Rehmannia glutinosa Libosch.HPLC were used for the deterimination of the fingerprints peaks relative to the content of catalpol. UV were used for the deterimination of the content of polysaccharides.Results The ethanol separating composition can remarkably decrease both the bleeding time and the clotting time of the mice.The separated composition by aqueous solution,ethanol 30%,ethanol 80% can also decrease the bleeding time and the clotting time.Dosages of aqueous solution were positively related to hemostatic effect respectively.Dose-effect relationship were unapparent in ethanol 30%.Dosages of ethanol 80% were negtively re-lated to hemostatic effect. Fifteen characteristic peaks were indicated in HPLC fingerprints. The relative retention time is as of the common peaks were also determined. The best extraction process is as follow:boiling the herbs with 12 times water and three times and every time for 1.5 hours,soaking for 15 minutes. The thin-layer chromatography spots were clear;and glucose was linear in the range of 0.0067～0.067mg/ml;and catalpol was linear in the range of 0.13～2.6μg.Conclusion The main substance treating hemostatic effect in Rehmannia glutinosa Libosch exists in the separated composition which eluted by Ethanol .The polysaccharides and total secoiridoid glycoside were the effective hemostatic components.This method is simple and accurate with a good reproducibility and provides a reference standard for the quality control of Rehmannia glutinosa Libosch. This extraction process is stable,reasonable and practical,providing a theoretical basis for industrial production. Through the inspection targets,the preparation of this is reasonable and feasible,the quality of products is basically stable. This method is simple,accurate,reliable and can be used as a quality control method for Rehmannia glutinosa Libosch Foam-rubber. |
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