Optimization of an Extraction Process to Obtain a Food-Grade Sulforaphane-Rich Extract from Broccoli (Brassica oleracea var. italica).

Sulforaphane (SFN) is a powerful health-promoting compound found in broccoli in the form of its inactive precursor, glucoraphanin (GFN). SFN formation occurs through the enzymatic hydrolysis of glucoraphanin by myrosinase under specific chemical conditions. Its incorporation in food formulations has been hindered by the thermal instability of SFN and low concentration in Brassicaceae. Then, extracting SFN from broccoli at a temperature below 40 °C appears as an option to recover and stabilize SFN, aiming at delivering it as a nutraceutical. We studied an eco-friendly extraction process to obtain an SFN-rich extract from broccoli. The effect of the broccoli mass/solvent ratio, ethanol concentration in the extractant solution, and extraction time on the recovery of SFN, GFN, phenolic compounds, and antioxidant activity were studied through a Box-Behnken design. The regression models explained more than 70% of the variability in the responses, adequately representing the system. The experimental factors differently affected the bioactive compound recovery and antioxidant activity of the extracts. The extraction conditions that allowed the highest recovery of bioactive compounds and antioxidant activity were identified and experimentally validated. The results may provide the basis for the design of a process to produce a sulforaphane-rich food supplement or nutraceutical by using a GRAS extractant.

Identification of Phytochemical-Based ß-Catenin Nuclear Localization Inhibitor in NSCLC: Differential Targeting Population from Member of Isothiocyanates.

Decades of research has convinced us that phytochemical compounds contained within the plant products are the real deal, and they provide benefits such as health maintenance an d cure to illnesses. One of the deadliest noncommunicable diseases today is lung cancer, hence its disease management still deserves attention. Wnt/ß-catenin pathway activation conferring cancer stem cell (CSC) activities to non-small cell lung carcinomas (NSCLCs) may explain why the disease is still difficult to cure. In the present study, we assessed several representatives of phytochemical categories consisting of alkaloids, chalcones and isothiocyanates for their inhibitory activity to nuclear localization of ß-catenin-an important event for Wnt/ß-catenin pathway activation, in lung cancer cell lines. Real-time cell analyzer confirmed that evodiamine (EVO), chelidonine (CHE), isoliquiritigenin (ISO), licochalcone-A (LICO), benzyl isothiocyanate (BI) and phenethylisothiocyanate (PI) exhibited anti-proliferative activities and cytotoxicities to adenocarcinoma cell line SK-LU-1 and human lung CSC primary cell line (HLCSC). Immunofluorescence assay identified that CHE, ISO, LICO, BI and PI were capable of reducing the number of cells harboring ß-catenin within the nuclei of these cells. We extended the characterizations of BI and PI in Wnt-dependent squamous cell carcinoma cell line NCI-H1703 on several CSC functions and found that BI was better at inhibiting soft agar colony formation as an output of self-renewal ability, whereas PI was more effective in inhibiting the growth of multicellular tumor spheroid model mimicking micrometastases. Both however were not able to inhibit migration and invasion of NCI-H1703. In conclusion, BI could potentially be used as a safer alternative to target undifferentiated CSCs as adjuvant therapy, whereas PI could be used as chemotherapy to remove bulk tumor.

Consumers acceptance and volatile profile of wall rocket (Diplotaxis erucoides).

Wall rocket (Diplotaxis erucoides) is a wild edible herb traditionally consumed in the Mediterranean regions with a characteristic, pungent flavour. However, little is known about its acceptance as a potential new crop. In the present study, an hedonic test with 98 volunteers was performed in order to evaluate the potential of wall rocket as a new crop. Three products were tested corresponding to microgreens, seedlings and baby-leaves. The volatile constituents were also studied due to their probable influence on acceptance, and compared to Dijon's mustard and wasabi. The degree of acceptance was mainly related to taste and pungency. Microgreens were well accepted, whereas seedlings and baby-leaves were mainly appreciated by individuals that enjoy pungent tastes. The purchase intent was also highly related to the acceptance of taste and pungency. The volatiles profile revealed that wall rocket was rich in allyl isothiocyanate, like mustard and wasabi. This compound may be greatly responsible of the relationship between the acceptance of mustard, wasabi and wall rocket. Microgreens displayed the highest levels of isothiocyanates, although the quantity of product tested by panellists did not probably allow the appreciation of such compounds. In baby-leaves, a significant decrease in isothiocyanates GC area and relative abundances was observed. These results suggest that wall rocket microgreens would be accepted by a significant proportion of the general public since pungency is lowly perceived in the product, despite its high levels of isothiocyanates. By contrast, baby-leaves may become a crop for a cohort of consumers that enjoy pungent flavours.

Phytochemical and Health-Beneficial Progress of Turnip (Brassica rapa).

Despite the advancement of medical science, diseases are part-and-parcel of human life. Plants have provided humans with medicines since time immemorial, and are still one of the primary sources for drug discovery. Brassica rapa L., commonly known as turnip, is one of the world's oldest cultivated vegetables. Besides being an important vegetable and source of oil, turnip is also used as a traditional medicine for the treatment of headaches, chest complaints, rheumatisms, oedemas, gonorrhoea, syphilis, and rabies. Glucosinolates and isothiocyanates (mainly 2-phenylethyl, 4-pentenyl, and 3-butenyl derivatives) are the main constituents of turnip with diverse bioactivities, especially for the protective effect against cancers. Besides, flavonoids, phenolics, indoles and volatiles are also concomitant in this plant. Pharmacological investigation on turnip revealed the antitumor, antihypertensive, antidiabetic, antioxidant, antiinflammatory, hepatoprotective, and nephroprotective effects. The anticancer property was found to be the most promising biological activity of turnip with 2-phenylethyl isothiocyanate, phenylpropionitrile, brassicaphenanthrene A, 6-paradol, and trans-6-shogaol as the major active constituents. Flavonoids and phenolics with high free radical scavenging activity should be corresponding to the antioxidant effects. Arvelexin, an indole derivative in turnip, was reported with various effects involving antiinflamatory, antihypertensive and hypolipidemic potency. In spite of many studies concerning either the chemical constituents or the biological activities of turnip, only a few cases disclosed the active ingredients responsible for diverse bioactivities. This review summarizes the research progress on the chemistry and health-benefits of turnip over the past 20 years to provide a reference for the further investigation.

Applying the hydrodistillation process to Pentadiplandra brazzeana Baill. root: a chemical assessment.

A chemical study of the volatile components obtained by applying the hydrodistillation and reflux processes to Pentadiplandra brazzeana roots was performed by GC-FID and GC-MS. The hydrodistillation process showed a total yield of 0.97% with 0.11% of essential oil and 0.86% of volatile compounds from the aqueous reaction medium; in the reflux process, the volatile extract yield was 1.03%. Benzylic-type isothiocyanates were the major degradation products of glucosinolates in the essential oil (95.0%); the CH2Cl2 extracts obtained from the aqueous solutions were characterised by alcohols and amines in both processes. This study has shown that during hydrodistillation, only 10% of the glucosinolate degradation products are recovered in the essential oil whereas 90% remain in the aqueous medium, being converted into alcohols and amines. The relative percentages of the different chemical classes recovered in our experimental conditions are discussed in relation with the glucosinolate composition in the raw material.

The primary active components, antioxidant properties, and differential metabolite profiles of radish sprouts (Raphanus sativus L.) upon domestic storage: analysis of nutritional quality.

BACKGROUND: This study aimed to analyze the nutritional quality of radish sprouts (Raphanus sativus L.) after domestic short-term storage. We stored fresh radish sprouts at 25 ± 1 °C and at 4 ± 1 °C for 12 h, and detected phenolic substances, glucosinolates, isothiocyanates, vitamin C, and various antioxidant and abiotic stress-related factors. We investigated nutrient-related metabolic differences and associated pathways and postharvest treatment effects on nutritional quality using metabolomic analysis. RESULTS: Most active substances and antioxidant properties, but not phenolic acids and vitamin C, decreased significantly (P < 0.05) upon domestic storage; this reduction decelerated at low temperatures. Short-term storage disrupted redox balance; low temperature enhanced stress resistance. Differences were observed in amino acid and vitamin derivatives, phospholipid accumulation, and organic acids. Short-term storage at ambient temperature promoted lysine, threonine, cysteine, vitamin H, phospholipid, and lauric (dodecanoic) acid accumulation, inhibiting proline, phosphatidic acid (PA) (14:1(9Z)/12:0), and phosphatidylcholine (PC) (O-18:0/O-18:0) accumulation; low-temperature, short-term storage promoted myristic acid and phospholipid accumulation and reduced methionine synthesis and vitamin H and K accumulation. CONCLUSION: Overall, the nutritional quality of radish sprout decreased upon short-term storage, with differences in certain active substances. © 2018 Society of Chemical Industry.

Development of validated high-temperature reverse-phase UHPLC-PDA analytical method for simultaneous analysis of five natural isothiocyanates in cruciferous vegetables.

In the present study reverse-phase UHPLC-PDA technique was developed at 60°C for simultaneous quantification of allyl, 3-butenyl, 4-(methylthio)butyl, benzyl and phenethyl isothiocyanates. The validation parameter showed a very good linearity, with a correlation coefficient of 1.00 for all detected standard analytes. Also, high precision and accuracy were observed with lowest obtained values of 1.39% and 99.1%, respectively. Different varieties of three plants, viz. Brassica rapa var. rapa L., Raphanus sativus L. var. oleiformis Pers. and Eruca sativa Mill., were analyzed with this method. After analysis, 4-(methylthio)butyl isothiocyanate was observed to be the major component in the varieties of arugula. Allyl, benzyl and phenethyl isothiocyanates were detected in turnip varieties and, in addition, 3-butenyl isothiocyanate was detected in radish varieties.

Production of phytotherapeutics from broccoli juice by integrated membrane processes.

An integrated membrane process for the recovery and concentration of phytotherapeutics in broccoli juice was investigated to develop a natural product that could be of interest for food and/or pharmaceutical industry. Following a pretreatment step to remove suspended solids, the juice was ultrafiltered through a 50kDa MWCO polyethersulfone membrane. The permeate stream was further concentrated by osmotic distillation up to 42.6°Brix. During ultrafiltration, a complete recovery of sulforaphane, malic acid and citric acid was achieved, where total phenolic content and ascorbic acid were recovered at a ratio of 94.5 and 92.4%, respectively. All the bioactive compounds identified in broccoli juice was well preserved during subsequent 6-fold concentration by osmotic distillation. No significant difference was observed in total antioxidant activity of the juice throughout the operation. The proposed integrated membrane process has shown promising potential to produce a natural concentrate enriched in phytotherapeutics of the broccoli juice.

Targeting Colorectal Cancer Proliferation, Stemness and Metastatic Potential Using Brassicaceae Extracts Enriched in Isothiocyanates: A 3D Cell Model-Based Study.

Colorectal cancer (CRC) recurrence is often attributable to circulating tumor cells and/or cancer stem cells (CSCs) that resist to conventional therapies and foster tumor progression. Isothiocyanates (ITCs) derived from Brassicaceae vegetables have demonstrated anticancer effects in CRC, however little is known about their effect in CSCs and tumor initiation properties. Here we examined the effect of ITCs-enriched Brassicaceae extracts derived from watercress and broccoli in cell proliferation, CSC phenotype and metastasis using a previously developed three-dimensional HT29 cell model with CSC-like traits. Both extracts were phytochemically characterized and their antiproliferative effect in HT29 monolayers was explored. Next, we performed cell proliferation assays and flow cytometry analysis in HT29 spheroids treated with watercress and broccoli extracts and respective main ITCs, phenethyl isothiocyanate (PEITC) and sulforaphane (SFN). Soft agar assays and relative quantitative expression analysis of stemness markers and Wnt/ß-catenin signaling players were performed to evaluate the effect of these phytochemicals in stemness and metastasis. Our results showed that both Brassicaceae extracts and ITCs exert antiproliferative effects in HT29 spheroids, arresting cell cycle at G2/M, possibly due to ITC-induced DNA damage. Colony formation and expression of LGR5 and CD133 cancer stemness markers were significantly reduced. Only watercress extract and PEITC decreased ALDH1 activity in a dose-dependent manner, as well as ß-catenin expression. Our research provides new insights on CRC therapy using ITC-enriched Brassicaceae extracts, specially watercress extract, to target CSCs and circulating tumor cells by impairing cell proliferation, ALDH1-mediated chemo-resistance, anoikis evasion, self-renewal and metastatic potential.

Identification of Glucosinolates in Seeds of Three Brassicaceae Species Known to Hyperaccumulate Heavy Metals.

Plants from the Brassicaceae family are known to contain secondary metabolites called glucosinolates. Our goal was to establish by LC/MS the glucosinolate profile of seeds of three Brassicaceae species known to hyperaccumulate heavy metals. We investigated Alyssum fallacinum auct. non Hausskn., Iberis intermedia Guers., and Noccaea caerulescens (J. Presl & C. Presl) F. K. Mey. Our results indicate that A. fallacinum seeds contain glucoiberin and glucoibervirin, which had not been previously identified in this plant. Furthermore, we report for the first time the presence of glucoiberin, glucoibervirin, glucotropaeolin, and sinigrin in I. intermedia. We have detected for the first time glucoconringiin in N. caerulescens. In addition, glucosinalbin, 4-hydroxyglucobrassicin, and glucomoringin were also detected.

Influence of Sulforaphane Metabolites on Activities of Human Drug-Metabolizing Cytochrome P450 and Determination of Sulforaphane in Human Liver Cells.

The influence of metabolites of sulforaphane, natural compounds present in broccoli (Brassica oleracea var. botrytis italica) and in other cruciferous vegetables, on drug-metabolizing cytochrome P450 (CYP) enzymes in human liver microsomes and possible entry of sulforaphane into human hepatic cells were investigated. Metabolites studied are compounds derived from sulforaphane by the mercapturic acid pathway (conjugation with glutathione and by following reactions), namely sulforaphane glutathione and sulforaphane cysteine conjugates and sulforaphane-N-acetylcysteine. Their possible effect on four drug-metabolizing CYP enzymes, CYP3A4 (midazolam 1'-hydroxylation), CYP2D6 (bufuralol 1'-hydroxylation), CYP1A2 (7-ethoxyresorufin O-deethylation), and CYP2B6 (7-ethoxy-4-(trifluoromethyl)coumarin O-deethylation), was tested. Inhibition of four prototypical CYP activities by sulforaphane metabolites was studied in pooled human liver microsomes. Sulforaphane metabolites did not considerably affect biological function of drug-metabolizing CYPs in human liver microsomes except for CYP2D6, which was found to be inhibited down to 73-78% of the original activity. Analysis of the entry of sulforaphane into human hepatocytes was done by cell disruption by sonication, methylene chloride extraction, and modified high-performance liquid chromatography method. The results have shown penetration of sulforaphane into the human hepatic cells.

Myrosinase Compatible Simultaneous Determination of Glucosinolates and Allyl Isothiocyanate by Capillary Electrophoresis Micellar Electrokinetic Chromatography (CE-MEKC).

INTRODUCTION: The functional food Cruciferous vegetables contain glucosinolates which are decomposed by the myrosinase enzyme upon tissue damage. The isothiocyanates are the most frequent decomposition products. Because of their various bioactivities, these compounds and the myrosinase is of high interest to many scientific fields. OBJECTIVE: Development of a capillary electrophoresis method capable of myrosinase-compatible, simultaneous quantification of glucosinolates and isothiocyanates. METHODS: Capillary electrochromatography parameters were optimised, followed by optimisation of a myrosinase-compatible derivatisation procedure for isothiocyanates. Vegetable extracts (Brussels sprouts, horseradish, radish and watercress) were tested for myrosinase activity, glucosinolate content and isothiocyanate conversion rate. Allyl isothiocyanate was quantified in some food products. RESULTS: The method allows quantification of sinigrin, gluonasturtiin and allyl isothiocyanate after myrosinase compatible derivatisation in-vial by mercaptoacetic acid. The chromatograhpic separation takes 2.5 min (short-end injection) or 15 min (long-end injection). For the tested vegetables, measured myrosinase activity was between 0.960-27.694 and 0.461-26.322 µmol/min/mg protein, glucosinolate content was between 0-2291.8 and 0-248.5 µg/g fresh weight for sinigrin and gluconastrutiin, respectively. The possible specificity of plants to different glucosinolates was also shown. Allyl isothiocyanate release rate was different in different vegetables (73.13 - 102.13%). The method could also be used for quantification of allyl isothiocyanate from food products. CONCLUSIONS: The presented capillary electrophoresis method requires a minimal amount of sample and contains only a few sample preparation steps, and can be used in several applications (glucosinolate determination, myrosinase activity measurement, isothiocyanate release estimation). Copyright © 2016 John Wiley & Sons, Ltd.

Integrated Process for Production of Galangal Acetate, the "Wasabi-Like" Spicy Compound, and Analysis of Essential Oils of Rhizoma Alpinia officinarum (Hance) Farw.

Rhizoma Alpinia officinarum (Hance) Farw, Zingiberaceae (AO), a ginger family herb exhibiting stimulant and a carminative bioactivity, is widely used in European and Asian countries as spicy condiment and medicinal uses. Allyl isothiocyanate (AITC) is the main pungent taste of native Wasabi (Wasabia japonica). The cytotoxicity of AITC has been implicated in thymus, adrenals, and white blood cells. Considering food safety, apparently a safer substitute for wasabi is worthy commercialized. Previously, we found AO crude paste to be rather feasible for use as a "Wasabi-substitute" in fresh meat and cold salads. A process linking cold ethyl acetate (EtAc) extraction with silica gel adsorption and reversed phase high-performance liquid chromatography (RP-HPLC) (mobile phase, 75% methanol) was used to isolate galangal acetate, the Wasabi-like taste constituent. AO contained abundant galangal acetate (3.84 ± 0.07%) compared to A. galangal (0.57 ± 0.16%), and as already confirmed by thin layer chromatography (TLC), gas chromatography (GC)/mass spectrometry (MS)/MS and Nuclear Magnetic Resonance Spectroscopy (NMR), galangal acetate was particularly thermally labile. The steam distilled essential oil (SDEO) of AO (0.14% on wet basis) contained 80 compounds (number of component, %): monoterpene hydrocarbon (21, 13.83%); oxygenated monoterpene (17, 27.08%); sesquiterpene hydrocarbon (20, 31.03%), and oxygenated sesquiterpene (20, 21.85%), respectively. However, no spicy wasabi-like constituent remained in SDEO. Alternatively, n-hexane, EtAc, and methanol extracts of AO all showed potent DPPH- and superoxide anion-scavenging activity. Conclusively, SDEO although contains 80 volatiles, galangal acetate is absent due to thermal instability. Galangal acetate exhibits pleasant "Wasabi-like taste" for which we have successively developed an integrated process for mass production.

Trace analysis of sulforaphane in bee pollen and royal jelly by liquid chromatography-tandem mass spectrometry.

In this study, we investigate for the first time the presence of sulforaphane (SFN) residues in two of the most currently consumed food/dietary supplements, royal jelly and bee pollen. Chromatography-tandem mass spectrometry (LC-MS/MS) was the method employed, the mass spectrometer consisting of an ion-trap mass analyzer used with electrospray ionization (ESI) in positive ion mode. An efficient sample treatment involving a solvent extraction with methanol, centrifugation, and concentration in a rotary evaporator was proposed. In all cases average analyte recoveries were between 92 and 106%. Chromatographic analysis (16min) was performed on a core-shell technology based column (Kinetex C18, 150×4.6mm, 2.6µm, 100Å). The mobile phase consisted of 0.02M ammonium formate in water and acetonitrile, with a flow rate of 0.5mL/min in gradient elution mode. The fully validated method was selective, linear from 8 to 1000µg/kg (bee pollen), or from 10 to 1250µg/kg (royal jelly), precise and accurate; relative standard deviation (% RSD) and relative error (% RE) values were below 10%. Low limits of detection (LOD) and quantification (LOQ) were obtained, namely, 3µg/kg (LOD) and 8 (bee pollen) and 10 (royal jelly) µg/kg (LOQ). The method was applied for SFN analysis in several royal jelly and bee pollen samples. SFN was detected at trace levels in some bee pollen samples (<23µg/kg) examined, whilst SFN went undetected in the royal jelly samples analyzed.

Effect of Se treatment on glucosinolate metabolism and health-promoting compounds in the broccoli sprouts of three cultivars.

Broccoli sprouts are natural functional foods for cancer prevention because of their high glucosinolate (GSL) content and high selenium (Se) accumulation capacity. The regulation mechanism of Se on GSL metabolism in broccoli sprouts was explored. In particular, the effects of Se treatment (100 µmol/L selenite and selenate) on the Se, sulfur (S), glucosinolate and sulforaphane contents; myrosinase activity and health-promoting compounds (ascorbic acid, anthocyanin, total phenolics and flavonoids) of three, 5 day old, cultivars were investigated. The treatment did not influence the total GSL and ascorbic acid contents; significantly increased the myrosinase activity and sulforaphane, anthocyanin and flavonoids contents; and decreased the total phenolics content. The increase in sulforaphane during early growth can be primarily attributed to the increased myrosinase activity caused by Se treatment. Broccoli sprouts with suitable selenite and selenate concentrations, in the early growth days, could be desirable for improved human health.

Inhibition of growth and induction of apoptosis in A549 cells by compounds from oxheart cabbage extract.

BACKGROUND: Oxheart cabbage (Brassica oleracea var. capitata) is a member of the Brassica genus. Although some studies on the anticancer effects of extracts from oxheart cabbage have been reported, comprehensive information on the bioactive fractions and components from oxheart cabbage extracts is still lacking. The aim of this study was to isolate and identify the bioactive fractions and components from oxheart cabbage seeds using activity-guided isolation methods. RESULTS: The cytotoxicity and apoptotic effects of fraction II, fraction III, iberverin, sulforaphane and iberin from oxheart cabbage seed extract were investigated. The results showed that all five components had inhibitory effects on the in vitro growth of A549 cells which were dose-dependent. These compounds also changed the morphology of A549 cells, and their inhibitory activity on A549 cells was as follows: sulforaphane > iberin > iberverin > fraction III > fraction II. The IC50 values were 3.53 ± 0.63, 4.93 ± 1.02, 7.07 ± 0.51, 15.56 ± 0.24 and 27.32 ± 0.63 µg mL(-1) respectively. Fraction II, fraction III, iberverin, sulforaphane and iberin induced cell apoptosis by increasing early apoptosis and late apoptosis/necrosis, and activation of caspase-3, -8 and -9. CONCLUSION: These results indicated that the decrease in A549 cell viability by active compounds from oxheart cabbage seed extract was due to the induction of apoptosis. © 2015 Society of Chemical Industry.

Effect of Essential Oils on Germination and Growth of Some Pathogenic and Spoilage Spore-Forming Bacteria.

The use of essential oils as a food preservative has increased due to their capacity to inhibit vegetative growth of some bacteria. However, only limited data are available on their effect on bacterial spores. The aim of the present study was to evaluate the effect of some essential oils on the growth and germination of three Bacillus species and Geobacillus stearothermophilus. Essential oils were chemically analyzed using gas chromatography and gas chromatography coupled to mass spectrometry. The minimal inhibitory and bactericidal concentrations of vegetative growth and spore germination were assessed using the macrodilution method. Germination inhibitory effect of treated spores with essential oils was evaluated on solid medium, while kinetic growth was followed using spectrophotometry in the presence of essential oils. Essential oil from Drypetes gossweileri mainly composed of benzyl isothiocyanate (86.7%) was the most potent, with minimal inhibitory concentrations ranging from 0.0048 to 0.0097 mg/mL on vegetative cells and 0.001 to 0.002 mg/mL on spore germination. Furthermore, essential oil from D. gossweileri reduced 50% of spore germination after treatment at 1.25 mg/mL, and its combination with other oils improved both bacteriostatic and bactericidal activities with additive or synergistic effects. Concerning the other essential oils, the minimal inhibitory concentration ranged from 5 to 0.63 mg/mL on vegetative growth and from 0.75 to 0.09 mg/mL on the germination of spores. Spectrophotometric evaluation showed an inhibitory effect of essential oils on both germination and outgrowth. From these results, it is concluded that some of the essential oils tested might be a valuable tool for bacteriological control in food industries. Therefore, further research regarding their use as food preservatives should be carried out.

Investigation of total phenolic, total flavonoid, antioxidant and allyl isothiocyanate content in the different organs of Wasabi japonica grown in an organic system.

BACKGROUND: This study was carried out to investigate the total polyphenol (TP), total flavonoid (TF), antioxidative effect and allyl isothyocyanate (ITC) content in different organs of wasabi plant grown in an organic system. MATERIALS AND METHODS: Invitro study of methanol and boiled water extracts of wasabi were conducted by analyzing the 1-1-diphenyl-2-picryl hydrozyl (DPPH) radial scavenging activity, metal chelating activity and total antioxidant capacity in a comparative manner. RESULT: The result revealed that methanol extract showed higher TP in flower (3644 mg TAE/100 g dw), leaf (3201 mg TAE/100 g dw) and fruit (3025 mg TAE/100 g dw) as compared to the boiled water extract. Similarly, TF content was also higher in methanol extracts of flower (1152 mg QE/100 g dw) and leaf (325 mg QE/100 g dw), however, the other parts showed ignorable value. RESULTS: Results of antioxidant activity were found at different magnitude of potency. The methanol extract of different parts of wasabi exhibited higher activity in total antioxidant capacity and DPPH radical scavenging assay as compared to water extract. In metal chelating assay, the boiled water extracts of leaf showed higher (76.9%) activity, followed by fruit (68.8%) and flower (62.8%). Ally ITC detected by gas chromatography was present in all of the tissues of wasabi plant but the content was found to be varied in different tissues. CONCLUSIONS: Overall, this study will allow consumers and processors to understand the possibility for medical application of wasabi plant by knowing the level of total polyphenol distribution, Ally ITC content and antioxidant property distributed in different parts and tissues.Key words: Allyl ITC, antioxidant, flavonoid, polyphenol, Wasabi japonica.

Concentration- and time-dependent effects of isothiocyanates produced from Brassicaceae shoot tissues on the pea root rot pathogen Aphanomyces euteiches.

Isothiocyanates (ITCs) hydrolyzed from glucosinolates (GSLs) in Brassicaceae tissue are toxic to soil organisms. In this study, the effect of aliphatic and aromatic ITCs from hydrated dry Brassicaceae shoot tissues on the mycelium and oospores of the pea root rot pathogen Aphanomyces euteiches was investigated. The profile and concentrations of GSLs in two test Brassicaceae species, Sinapis alba and Brassica juncea, and the ITCs from the dominant hydrolyzed parent GSLs were monitored. The concentrations of dominant ITCs and pathogen exposure time were evaluated in in vitro experiments. The greatest effect on the pathogen was observed from aliphatic ITCs hydrolyzed from B. juncea tissue, and the effect depended on the ITC concentration and exposure time. ITCs were more effectively hydrolyzed from B. juncea GSLs than from S. alba GSLs; i.e., the ITC/GSL ratio was higher in B. juncea than in S. alba tissue, giving a different release pattern. The release of phenylethyl isothiocyanate, which was common to both species, followed a pattern similar to that of the dominant ITC in each crop species. This suggests that traits other than GSL content, e.g., plant cell structure, may affect the release of ITCs and should therefore influence the choice of species used for biofumigation purposes.