Studies On Tablet Of Semen Sojae Preparatrm Isoflavone Preparation

Objective:Semen Sojae Preparatrm is a general all purpose traditional chinese drug. It has the effects of inducing diaphoresis, relieving restlessness,and dispelling emissionhcat heat. It is used to treat fiu, heedaehe, fidgeting due to deficiency and insomnia, etc. Isoflavones is major active component in fermented soybean chemical composition,which has function like phytoestrogens. Our earlier research shows that isoflavones has a therapeutic effect on menopausal women's cardiovascular diseases. These findings are experimental and theoretical bases to develop resources of semen sojae preparatrm and prepare a new drugs for menopausal women's cardiovascular diseases. This research optimize the extraction and emundation technology of isoflavones in fermented soybean. Furthermore, in conjunction with the powder characteristics, we rudimently approached shaping technology of soybean isoflavones.Methods:1 Studies on determination methodswe performed ultraviolet spectrophotometry to establish the determination methods of total isoflavones and HPLC method to establish the determination methods of genistein, as well as the methodology. 2 Studies on the process of extraction, separation, purificationwe used the content of total isoflavones and genistein as indicators to select the best extraction solvency, through single factor extraction solvency inspection; we adopted four factors, three levels orthogonal design to optimize the extraction process of total isoflavones, furthermore, in the combination of the actual extraction process for the production with the verification test, through the verification test, we identified the extraction process of total isoflavones.Macroporous resin was used for the separation and purification of total isoflavones. With static and dynamic absorption method, respectively, we inspected different models and different manufacturers macroporous resin, and selected the optimum macroporous resin for total isoflavones isolation and purification. We then inspected the impect factors of macroporous resin absorption and elution rate: the concentration of liquid , sample flow rate, the largest sample volume, eluting water amount, elution ethanol concentration, eluting ethanol amount and elution velocity, from which we got the process conditions by macroporous resin to separate and purify total isoflavones.3 Studies on preparation process of taballa of total isoflavones The prescription of the taballa total isoflavones was determined by comparison of different adhesives, disintegrants and lubricants. Ethanol concentration(75%,80%,85%,95%), PVP concentration (1%,3%,5%,10%), sodium carboxymethyl cellulose amount and magnesium stearate amount were investigated to determine the process of total isoflavones taballa. Some factors such as slick, disintegration and friability affecting the quality of the taballa were considered as well.Results:1 The ultraviolet spectrophotometry for the determination of total isoflavones and HPLC methods for the determination of genistein were established.1.1 Ultraviolet spectrophotometry for the determination of total isoflavonesReference standard solution preparation: precision take appropriate amounts of genistein reference substance and put it into methanol to make it containing 3.864μg genistein per ml solution.Test solution preparation: precisely draw semen sojae preparatrm extracting solution 5 ml, put it into a pan, and make it dry, add up 15 ml water to solve the residue, extract isoflavones with acetidin from water solution 6 times, and toties quoties 25 ml.then combine acetidin layer, and make it dry, add up methanol to solve the residue into 50 ml volumetric flask and dilute to scale, and then shake up evenly, standby.Selection of detection wavelength: we scan standard solution and test solution with diode array detector between 200nm and 700nm. Both have maximum absorption wavelengh at 261 nm. So 261nm was determined as the detection wavelength. Determination: determine the content of the test solution and reference substance solution with ultraviolet spectrophotom- eter.1.2 HPLC method for the determination of genisteinChromatographic conditions and system suitability test: 18 alkyl silane bonded silica was used as filler and methanol-0.1% glacial acetic acid (53:47) was used as the mobile phase, the detection wavelength is 261nm, and the flow rate is 1ml/min. Theoretical plate number calculated by genistein should not be lower than 3000.Reference standard solution preparation: precision take appropriate amounts of genistein reference substance and put it into MeOH to make it containing 9.92μg genistein per ml solution, standby.Test solution preparation: draw semen sojae preparatrm extracting solution, and filter it with 0.45μm filtration membrane, standby.Determination: draw the test solution and reference substance solution 10μl respectively, inject into HPLC to determine the content.2 The studies findings of the process of extraction, separation and purification2.1 the sudies finding of the process of extractionTo optimize semen sojae preparatrm extraction process by adopting the single factor inspection and orthogonal test: add 70% ethanol(8 times), extract three times, 2 hours pre time. condense liquid to 1ml liquid containing 0.1g herbs.2.2 The studies findings of the process of separation and purificationTo screen out the optimum macroporous resin(HPD500) by adopting static and dynamic adsorption methods to separate and purify total isoflavones in semen sojae preparatrm. its absorption capacity and washout rate is higher than other resins. The process to purify total isoflavones by HPD500 macroporous resin was defined as: add water to dilute to per ml medicine liquid containing 0.15g herbs, adopte 4BV/h through HPD500 macroporous resin column (maximum adsorption quantity was per ml macroporous resin adsorpting 1.17g herbs), use twelve times volume of water to wash firstly, following by fifteen times volume of 70% ethanol to wash and the elution velocity is 4BV/h, collect the elution liquid and dry it completely under lowered pressure, and then pulverize it into 80 oculus.3 the studies findings of the forming process of taballa total isoflavonesTo set up modeling technique of through research on taballa total isoflavones: the fine powder purified by HPD500 macroporous resin, starch, carboxymethylcellulose sodium, 5% PVP prepared by 80% ethanol were used to granulate. The bead was dried and completed with 20 oculus screen. Vitamin c and magnesium stearate were added to make the final tablla. Taballa were light brown and smooth indicating that the process is stable and feasible.Conclusions: The ultraviolet spectrophotometry method for determination of the content of total SSP isoflavones and HPLC method for genistein were established through investigation of the methodology. The results show that the method is stable and reliable and can be applied to assay total isoflavones and genistein. The optimal processes of extraction, separation and purification were as follows: add 70% ethanol(8 times), extract three times, 2 hours pre time, condense liquid to 1ml liquid containing 0.15g herbs, adopt 4BV/h through HPD500 macroporous resin column (maximum adsorption quantity was 1.17g herbs per ml macroporous resin), use twelve times volume of water to wash firstly, following fifteen times volume of 70% ethanol to wash and the elution velocity is 4BV/h, collect the elution liquid and concentrate it. Dry it completely and then pulverize it into 80 oculus. The optimal process of shaping tablla was as follows: the fine powder purified by HPD500 macroporous resin, starch, carboxymethylcellulose sodium, 5% PVP prepared by 80% ethanol were used to granulate. The bead was dried and completed with 20 oculus screen. Vitamin c and magnesium stearate were added to make the final tablla, which was qualified.