Sulforaphene and sulforaphane in commonly consumed cruciferous plants contributed to antiproliferation in HCT116 colon cancer cells

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.

Sulforaphene in Raphanus sativus L. var. caudatus Alef increased in late-bolting stage as well as anticancer activity

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.

Development and Application of an Extraction and Quantitative HPLC Method for Sulforaphene in Radish

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.

8种萝卜籽油的组成与抗氧化活性

用二氯甲烷作提取剂提取了富含异硫氰酸盐的萝卜籽油,并分析了其脂肪酸组成、莱菔素(sulforaphene)含量及抗氧化性能。结果表明:8种萝卜籽油得率为34.65%～37.91%;8种萝卜籽油中含有多种脂肪酸,其中芥酸含量最高,为29.45%～35.01%,其次是油酸(21.99%～29.59%)、亚油酸(9.53%～13.63%)和二十碳一烯酸(8.88%～10.52%),再次是α-亚麻酸、棕榈酸,含量分别为5.30%～8.35%、5.37%～5.92%,不饱和脂肪酸含量达88.08%～89.73%;8种萝卜籽油含有28.39～114.73 mg/kg的sulforaphene,其中白玉春的含量最高,春萝卜籽油比秋萝卜籽油含量相对高。FRAP法抗氧化测定中,盛丰白萝卜籽油的活性最高。

萝卜苗中异硫氰酸盐的提取工艺

以正己烷和二氯甲烷为溶剂,4-methylthio-3-butenyl isothiocyanate(MTBITC)和4-methylsulfinyl-3-butenyl isothiocyanate(Sulforaphene)的总量为提取效率研究萝卜苗中异硫氰酸盐的提取工艺。结果表明:提取萝卜苗中异硫氰酸盐的最佳工艺条件为自身酶解时间3～5min、液固比2:1(mL/g)、按正己烷-二氯甲烷-二氯甲烷的溶剂顺序各提取1次、提取时间3min/次。