The Preparation And Industrialization Of The High Purity Sulforaphane

Sulforaphane is considered as one of natural products which has wonderful effects on the prevention and treatment to cancer at present. It has been identified that sulforaphane could delay preneoplastic lesions and had a promising effect on many kinds of cancers involving in hepatic, breast, colon, gastric, etc. The preparation studies of sulforaphane are limited, while the most of studies are concerned to pharmacology and the mechanism. The preparation and indudtrialization of the high purity sulforaphane was investigated in this paper.Effect of grinding on the formation of sulforaphane in broccoli seeds was investigated. With decreasing the particle size of the broccoli seed powders, the water absorption capacity (from 0.335g/g to 0.544g/g), protein content (from 54.91mg/g to 231.83mg/g), glucoraphanin content (from 31.33mg/g to 40.36mg/g), the amount of glucose formed (78.00nmol to 200.53nmol) and the sulforaphane content (from 5.85mg/g to 14.04mg/g) were increased significantly. However, the myrosinase activity was not affected by grinding. The influence of the particle size and pH on the broccoli seed hydrolyzate filtration features was studied. The filtration characteristics of the hydrozate were measured; the specific cake resistance and the cake-layer resistance were calculated. The results indicated that 285.51μm had the best filtration characteristic and the specific cake resistance and cake-layer resistance were 2.15×1012m/kg and 15.97×1012 m-1, respectively. The next is 64.65μm; the specific cake resistance and cake-layer resistance were 3.45×10 m/kg and 26.76×1012 m-1, respectively. The optimum pH is 2.5-3.The purification of sulforaphane using preparation HPLC was scaled up.The separative conditions of three preparation columns (19×300mm, C18,15μm; 32×300mm, C18,50μm; 44×300mm, C18, 50μm) were groped. The purity of sulforaphane were 96.5%,87.7%, 85.5% and the recovery were 95.7%,92.1%,80.5%, respectively. The separative efficiency of the columns was also compared in this paper.The results indicated that the separative efficiency was decreased with the increase of the packing particle sizes and column diameters.The sulforaphane was also separation and purified by polystyrene-based resin reversed phase chromatography (10×300mm, 30μm). The separation conditions were groped and the optimum parameters were 30% acetonitrile in water as the mobile phase,2 ml/min of the flow-rate in isocratic elution mode. The highest purity of sulforaphane was over 90% at the condition of 20mg loading-amount. The separative efficiency was decreased with the increase of the loading amount. To raise production ability, preparative LC can be operated at the over-loading condition, and heart-cutting should be used for improving the purity of sulforaphane.