Sulforaphene, an isothiocyanate generated by the conversion of glucoraphenin in radish(Raphanus sativus L.), plays an important role in plants as a defensive compound and has been shown to have anti-cancer properties....Sulforaphene, an isothiocyanate generated by the conversion of glucoraphenin in radish(Raphanus sativus L.), plays an important role in plants as a defensive compound and has been shown to have anti-cancer properties. However, a reliable method for the determination of sulforaphene in radish has not yet been established. In the present study, the enzymolysis conditions, extraction solvent and extraction conditions employed to obtain sulforaphene from radish were all optimized using single factor experiments and orthogonal testing. During these trials, sulforaphene was separated and quantified using reverse phase C18 high performance liquid chromatography(HPLC), employing methanol︰water(3︰7 ν/ν) as the mobile phase at a flow rate of 0.3 mL · min-1 together with a detection wavelength of 245 nm. Evaluation of this newly established method showed good linearity between the sulforaphene concentration and the characteristic peak area over the range of 5 to 200 μg· m L-1(R2 = 0.9996). This method was thus found to be precise. It was established that sulforaphene was extracted from radish with 95.48% efficiency(RSD = 0.94%) using the optimized conditions. This same technique was successfully applied to the quantification of sulforaphene in the fleshy roots of different radish germplasms as a means of selecting the germplasms with the highest sulforaphene levels.展开更多
Objectives: To evaluate the concentration differences of sulforaphene and sulforaphane at various ages and in different parts of Raphanus sativus L. var. caudatus with respect to their potential cancer preventive effe...Objectives: To evaluate the concentration differences of sulforaphene and sulforaphane at various ages and in different parts of Raphanus sativus L. var. caudatus with respect to their potential cancer preventive effect on HCT116 colon cancer cells.Methods: FTIR–ATR and GC–MS were used to characterize the isothiocyanates in the plant extracts followed by HPLC for quantification. Antiproliferation and apoptosis induction were determined by using MTT assay and flow cytometry, respectively.Results: The respective rank of anticancer activity of Raphanus sativus were as follows:vegetative(3 week) < older rosette(4 week) < early-bolting(5 week) < senescence(7 week) < late-bolting(6 week). The low to high concentration of sulforaphene and sulforaphane occurred in the same stage order.Conclusions: The reproductive parts(flower, pod, and dry seed) of Raphanus sativus have the greatest isothiocyanate concentration, evidenced by a sulforaphene concentration higher than the sulforaphane. This result should inform the selection of the most appropriate harvesting stage and plant part for use as a potential chemopreventive agent.展开更多
Objective: To analyze two isothiocyanates(sulforaphene and sulforaphane) and their antiproliferative effect of 11 indigenous cruciferous vegetables.Methods: Phytoconstituents identification was conducted by high perfo...Objective: To analyze two isothiocyanates(sulforaphene and sulforaphane) and their antiproliferative effect of 11 indigenous cruciferous vegetables.Methods: Phytoconstituents identification was conducted by high performance liquid chromatography and gas chromatography-mass spectrometer techniques. The antiproliferation was evaluated in colon cancer cell line HCT116 by MTT assay.Results: Isothiocyanate identification by high performance liquid chromatography showed that broccoli, cabbage, "Khi-Hood"(Raphanus sativus L. var. caudatus Alef) and Chinese radish contained isothiocyanates sulforaphane. Sulforaphene and sulforaphane in broccoli, cabbage and "Khi-Hood" were characterized by the gas chromatography-mass spectrometer analysis. Antiproliferation screening by MTT assay found that the potent plants which possessed IC_(50) below 50 mg/m L were cabbage and "Khi-Hood", while the others had low antiproliferation with IC_(50) higher than 50 mg/m L. Difference in antiproliferation was probably due to difference existed phytochemical constituents in each plant. "Khi-Hood" possessed the highest antiproliferation against HCT116 with the lowest IC_(50)at(9.42 ± 0.46) mg/m L. The IC_(50) of chemotherapeutic drug(mitomycin C)was(19.12 ± 1.00) mg/m L, while both melphalan and 5-fluorouracil possessed the IC_(50) value higher than 50 mg/m L.Conclusions: Commonly consumed cruciferous vegetables exerted varied antiproliferation and isothiocyanate contents. High isothiocyanate content in "Khi-Hood" was contributed to high antiproliferation. Among 11 plants studied, "Khi-Hood" could be an alternative chemopreventive diet.展开更多
基金supported by the National Key Technology R&D Program of the Ministry of Science and Technology of China(2013BAD01B04-12012BAD02B01)the National High-tech R&D Program of China(2012AA021801-4)
文摘Sulforaphene, an isothiocyanate generated by the conversion of glucoraphenin in radish(Raphanus sativus L.), plays an important role in plants as a defensive compound and has been shown to have anti-cancer properties. However, a reliable method for the determination of sulforaphene in radish has not yet been established. In the present study, the enzymolysis conditions, extraction solvent and extraction conditions employed to obtain sulforaphene from radish were all optimized using single factor experiments and orthogonal testing. During these trials, sulforaphene was separated and quantified using reverse phase C18 high performance liquid chromatography(HPLC), employing methanol︰water(3︰7 ν/ν) as the mobile phase at a flow rate of 0.3 mL · min-1 together with a detection wavelength of 245 nm. Evaluation of this newly established method showed good linearity between the sulforaphene concentration and the characteristic peak area over the range of 5 to 200 μg· m L-1(R2 = 0.9996). This method was thus found to be precise. It was established that sulforaphene was extracted from radish with 95.48% efficiency(RSD = 0.94%) using the optimized conditions. This same technique was successfully applied to the quantification of sulforaphene in the fleshy roots of different radish germplasms as a means of selecting the germplasms with the highest sulforaphene levels.
基金partially supported by the Higher Education Research Promotion and National Research University Project of ThailandOffice of the Higher Education Commission,through the Food and Functional Food Research Cluster and Research and Development of Herbal Nutraceutics Subcluster of Khon Kaen University (No. F-2553-M-11 and NRU541051)the Khon Kaen University Research Fund (No. 564301)
文摘Objectives: To evaluate the concentration differences of sulforaphene and sulforaphane at various ages and in different parts of Raphanus sativus L. var. caudatus with respect to their potential cancer preventive effect on HCT116 colon cancer cells.Methods: FTIR–ATR and GC–MS were used to characterize the isothiocyanates in the plant extracts followed by HPLC for quantification. Antiproliferation and apoptosis induction were determined by using MTT assay and flow cytometry, respectively.Results: The respective rank of anticancer activity of Raphanus sativus were as follows:vegetative(3 week) < older rosette(4 week) < early-bolting(5 week) < senescence(7 week) < late-bolting(6 week). The low to high concentration of sulforaphene and sulforaphane occurred in the same stage order.Conclusions: The reproductive parts(flower, pod, and dry seed) of Raphanus sativus have the greatest isothiocyanate concentration, evidenced by a sulforaphene concentration higher than the sulforaphane. This result should inform the selection of the most appropriate harvesting stage and plant part for use as a potential chemopreventive agent.
基金Supported by Higher Education Research Promotion and National Research University Project of Thailand,Office of the Higher Education Commission,through the Food and Functional Food Research Cluster and Research and Development of Herbal Nutraceutics Subcluster of Khon Kaen University(F-2553-M-11 and NRU541051)
文摘Objective: To analyze two isothiocyanates(sulforaphene and sulforaphane) and their antiproliferative effect of 11 indigenous cruciferous vegetables.Methods: Phytoconstituents identification was conducted by high performance liquid chromatography and gas chromatography-mass spectrometer techniques. The antiproliferation was evaluated in colon cancer cell line HCT116 by MTT assay.Results: Isothiocyanate identification by high performance liquid chromatography showed that broccoli, cabbage, "Khi-Hood"(Raphanus sativus L. var. caudatus Alef) and Chinese radish contained isothiocyanates sulforaphane. Sulforaphene and sulforaphane in broccoli, cabbage and "Khi-Hood" were characterized by the gas chromatography-mass spectrometer analysis. Antiproliferation screening by MTT assay found that the potent plants which possessed IC_(50) below 50 mg/m L were cabbage and "Khi-Hood", while the others had low antiproliferation with IC_(50) higher than 50 mg/m L. Difference in antiproliferation was probably due to difference existed phytochemical constituents in each plant. "Khi-Hood" possessed the highest antiproliferation against HCT116 with the lowest IC_(50)at(9.42 ± 0.46) mg/m L. The IC_(50) of chemotherapeutic drug(mitomycin C)was(19.12 ± 1.00) mg/m L, while both melphalan and 5-fluorouracil possessed the IC_(50) value higher than 50 mg/m L.Conclusions: Commonly consumed cruciferous vegetables exerted varied antiproliferation and isothiocyanate contents. High isothiocyanate content in "Khi-Hood" was contributed to high antiproliferation. Among 11 plants studied, "Khi-Hood" could be an alternative chemopreventive diet.
