Studies On Technology Of Isolation And Purification Of Gulcoraphanin

Gulcoraphanin is hydrolysed to sulforaphane. Sulforaphane is the activecomponent which is thought to result in certain cancer protection. Most of theresearches of sulforaphane are directly from the gulcoraphanin. There is not highpurity gulcoraphanin in the market。Along with the deep research of glucosinolates,the demand of high purity gulcoraphanin is urgent.In this paper, gulcoraphanin is the object of the research. The raw extractionof gulcoraphanin was made from broccoli seeds, and then purified by an anionexchange column of DEAE-Sephadex A25. Pure glucoraphanin was obtained fromthe purified sample followed by preparative HPLC. The purity was identified byMS, NMR, IR and UV spectroscopy. The results were shown as follows:1. The optimal condition above glucosinolates extracted from in broccoliseeds: Using the single factor test and orthogonal design by the extraction volumeof glucosinolates optimize the effect of methanol, solvent concentration, extractiontemperature and extraction time. The result is70%methanol solution as extractant,1:15of the ratio of the solid to liquid,15min of extraction time,80℃ofextraction temperature, twice of extraction time. Under the optimal condition,glucosinolates content was74.483μmol/g in extract.2. The condition of glucoraphanin analysis by HPLC: The analyses wereperformed with UV detector using a Global ChromatographySP-120-5-C18-AP(5μm，4.6mm×200mm) column. The flow rate was1.0mL per minand the chromatograms were monitored at229nm. The mobile phase was0.1%formic acid dissolved in2%v/v acetonitrile/water. Glucoraphanin was identifiedby MS. The retention time of the number3chromatogram peak was approximately4min.3. The condition of glucoraphanin purification by preparative HPLC:Preparative HPLC were performed with UV detector using a GlobalChromatography SP-120-5-C18-AP(10μm，10mm×250mm) column. The flow ratewas4.0mL per min and the chromatograms were monitored at229nm. The mobilephase was0.1%formic acid dissolved in2%v/v acetonitrile/water. The injectionwas200μL.3. Identification about the composition: The purified composition is glucoraphanin identified by UV、IR、MS and NMR.