Extraction, Purification, Identification And Activity Study Of Active Compounds From Old Preserved Radish In Southern Fujian

Old preserved radish(Raphanus Sativus),produced by traditional method,is known as a characteristic of preserved food in Southern Fujian and Guangdong Province of China made by high quality early winter radish and bold marinated.Years of natural maturing and hidden in old altar induce its glossy black color and unique flavor.The aim of this study was to find some active compounds from old preserved radish and provide scientific evidence on its high food and therapeutic value.Firstly,the chemical compositions of old preserved radish in different years and fresh radish have been investigated by high performance liquid chromatography-diode array detection(HPLC-DAD),and the HPLC fingerprints of the methanol extracts were made.Traditional Chinese medicine(TCM)chromatographic fingerprint similarity evaluation system version A was applied to analyze and evaluate the similarities of the HPLC fingerprints.It concluded that old preserved radish in five-year presented large difference relatively.Secendly,the methonal and ethyl acetate extracts of five-year preserved radish were separated and purified by using Sephadex LH-20 resin,positive phase and reversed phase silica gel column(RP-C18)chromatography.Compound MA,MD,MQ and MI were purified from methona extract.Compound EU,EW,EX,EY,EZ,Ea and Eb were purified from ethyl acetate extract.The structures of compounds were assigned combined with thin layer chromatography(TLC),UV detection,electrospray ionization mass(ESI-MS),nuclear magnetic resonance(1H-NMR,13C-NMR,1H-1H COSY,DEPT135,HMBC and HSQC).Those compounds were determined as Linoleic acid methyl ester(MA),2,6-Dihydroxyacetophenone(MD),5-Hydroxymethylfurfural(MI),p-Hydroxybenzaldehyde(MQ),?-Sitosterol(EU),?-Sitosterol-D-glucopyranoside(EW),Di-2-ethylhexyl phthalate(EX),?-Linolenic acid(EY),1-Hexadecanoylpropan-2,3-diol(EZ)and chaenomic acid A(Ea).They were first discovery in preserved radish except p-Hydroxybenzaldehyde.2,6-Dihydroxyacetophenone,5-hydroxymethylfurfural,p-Hydroxybenzaldehyde,?-Sitosterol-D-glucopyranosidewere proved as new compoundsduring curing through comparing the HPLC charts of five-year preserved radish and freshradish.Finally,the antimicrobial activity and proliferation inhibition in vitro of compounds weretexted conbined with agar diffusion assay,broth microdilution method and MTT.2,6-Dihydroxyacetophenone,5-hydroxymethylfurfural,p-Hydroxybenzaldehyde have strong inhibition on Saccharomyces cerevisiae.Bacillus subtilis and Escherichia coli were extremely susceptible to 2,6-Dihydroxyacetophenone and p-Hydroxybenzaldehyde.5-hydroxymethylfurfural,p-Hydroxybenzaldehyde and p-hydroxyphenylethyl alcohol can inbihit the proliferation of Hep G2,whose IC50 were 44.72 ?g/m L,42.25 ?g/m L and 43.50?g/m L.Meanwhile,they have no effect on LO2.