| Soybean, the seed of Glycine max(L.)Merr of the Pea family, is one of the farmcrops widly planted in China. Recently the pharmacology action of isoflavones,especially the fade estrogen action ,were attrative. Soybean isoflavones is widly usedin medicine, cosmetics and food.In this paper, isoflavones was extracted by security solvent from soybeanresidue.Iosflavones with high purity and high additional value was get with noinfluence on the nutrient value of the soybean residue. The economic benefits of thesoybean residue was greatly improved .High purity isoflavones was greatlydemanded.The preparation of high purity isoflavones was studied in this paper as follows :1) A new method of high performance thin layer chromatographic (HPTLC) andthin layer chromatographic (TLC) to qualitatively analyze isoflavones were developed.The optimal conditions of thin layer chromatography were as follows: silica gelGF254 as stationary phase, chloroform-methanol-acetic acid(93:7:0.5,v/v/v)as the mobile phase, the Rfvalues of daidzein and genistein were 0.47 and0.58.2)The method which used thin layer chromatographic scanner to quantitativeanalyze isoflavones was developed.Use reflectometry single wavelenth to scan, λ=260nm. The calibration equation of daidzein is Y=120840.5X+1859.1, The relatedcoefficient is 0.998. The RSD is 1.72%. The average recovery is 96.3 %. Thecalibration equation of genistein is Y=108098X+17464.1, The related coefficient is0.999. The RSD is 1.55%. The linear range is 0.3μg to 3μg. The average recovery is98.4 %. The LOD of daidzein is 6.56×10-7μg,the LOD of genistein is 1.98×10-7μg.3)A method of the reverse phase high performance liquid chromatographic(HPLC) determination of isoflavones was developed. The optimal HPLC conditionswere as follows: Apollo C18 column as the stationary phase; gradient elution, Asolvent=1%acetic acid,B solvent=80%methanol+1%acetic acid;0min～40min ,80%B ～ 100 % B,1mL/min of flow rate;25 ℃ of column temperature; UVdetector(260nm). Under these conditions, a baseline separation of daidzein and igenistein was obtained, and the retention time of daidzein is 37.5 minutes. Thecalibration equation is Y=15762638X+185870. The related coefficient is 0.999. Thelinear range is 2μg to 10μg. The RSD is 1.12%. The average recovery is 98.1 %. Theretention time of genistein is 40 minutes.The calibration equation is Y =56746926X-87078. The related coefficient is 0.999. The linear range is 2μg to 10μg.The RSD is 1.41%. The average recovery is 98.5 %.4)The extraction and hydrolysis of isoflavones were studied. The extraction ofisoflavones was performed by ethanol aqueous by refluxing. The optimal of extractionwas as follows:The volume rate of raw stuff and 65% ethanol was 1:4;extraction times was 3;extraction time was 3h, extraction temperature was 60℃.The optimal conditions ofhydrolysis which were determined by orthogonal test was as follow: the volume rateof raw stuff and 1mol/LHCl was 1:2; the volume rate of raw stuff and acetic esterwas 1:2.5; hydrolysis time was 1h, hydrolysis temperature was 65℃.The absorptionof isoflavones by eight kinds of resins and polyamide resin were studied. Theabsorption capacity, adsorption rate and the static desorption curve were determined.6) High-purity daidzein and genistein sample was obtained by preparative thinlayer chromatography (PTLC) and preparative high performance liquidchromatography (PHPLC). The samples were identified as daidzein and genistein bymass spectrum .The components of isoflavones were analysied by HPLC.5) The paper preliminary study on the absorption from isoflavones by resin andstatic absorb research by eight kinds of resins and polyamide, confirm the absorbvolume, adsorption rate and static adsorption curve of different resins and polyamide7) Large-scale experiment. 50g high purity isoflavones, which purity is 80%, wasobtained from 100Kg soybean residue. The high purity isoflavones was added tocosmetics to generate...
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