Impact of different drying trajectories on degradation of nutritional compounds in broccoli (Brassica oleracea var. italica)

This work concerns the degradation of the nutritional compounds glucoraphanin (GR) and vitamin C (Vc), and the inactivation of the enzyme myrosinase (MYR) in broccoli (Brassica oleracea var. italica) during drying with air temperatures in the range of 30–60 °C. Dynamic optimization is applied to find the optimized temperature trajectories that minimize degradation and inactivation. Simulation and experimental results for optimized temperature trajectories are compared to constant inlet air temperature drying at 40 and 50 °C. The results show that with the optimized temperature trajectories the retention of GR, MYR and Vc improved significantly. Moreover, the experiments show that degradation and inactivation during drying is slower than expected from kinetic studies. The deviation is explained from the difference in the physical state of the samples used in the drying experiments, i.e. original plant tissue versus the grounded state of the plant tissue used in the experiments for the kinetic studies. The results indicate that besides temperature and moisture content the physical state is also an important aspect in the degradation of nutritional compounds and enzymes.     

Genotypic variation of glucosinolates in broccoli (Brassica oleracea var. italica) florets from China

Five main cultivars from China and 143 parent materials grown in a greenhouse were used to investigate the glucosinolates in broccoli florets. Eight aliphatic glucosinolates, four indole glucosinolates, and one aromatic glucosinolate were identified and quantified using high-performance liquid chromatography (HPLC). The results showed that glucoraphanin, glucobrassicin, 4-methoxyglucobrassicin and neoglucobrassicin were present in all samples. However, the predominant type of glucosinolate was different among pure lines. The anti-cancer glucoraphanin concentration ranged from 0.06 to 24.17 μmol/g in pure lines and from 1.57 to 5.95 μmol/g in commercial cultivars. The progoitrin concentration in commercial cultivars varied from 1.77 to 6.07 μmol/g with a mean value of 3.20 μmol/g. Significant variations were observed in the concentration of individual glucosinolates and in each class of glucosinolates among broccoli populations. Dozens of specific lines with altered glucosinolate profiles, as well as ten good candidates for breeding high-chemoprotective glucosinolate cultivars, were obtained according to the putative glucosinolate pathway in broccoli.     

Electron-beam irradiation of fresh broccoli heads (Brassica oleracea L. italica)

Broccoli is a popular item in the diet of the US population, commonly found in salads ready-to-eat. The recent recalls of fresh produce due to contamination with Escherichia coli and other pathogens emphasize the need to find effective means to treat minimally processed fresh foods. Our study assessed the effect of ionizing radiation using electron beams on the shelf-life, physicochemical properties, and consumer acceptability of broccoli florets. One-hundred broccoli heads were irradiated at 1, 2, and 3 kGy with a 10 MeV linear accelerator at 22 °C. We monitored pH, color, texture, respiration rate, weight loss, chlorophyll, total carotenoids, and vitamin C of irradiated and non-irradiated samples at 5-day intervals up to 14 days at 4 °C and 95% RH. Fifty consumer panelists scored the samples using a nine-point hedonic scale.Irradiation did not affect color, firmness, pH, and weight loss of the samples. Both irradiated samples and controls showed a slight change in color during storage, though the effect was not dose-dependent. Irradiation affected (P < 0.05) the respiration rates on the first 5 days of storage (higher CO₂ levels) but all samples, including the controls, attained the same equilibrium value. Vitamin C content of all samples decreased (P < 0.05) with storage time. Chlorophyll and total carotenoids content followed the same trend. In terms of overall acceptability, color, odor, and texture, all irradiated samples were highly accepted by the panelists with scores of 5 and above. By the end of shelf-life, only the controls showed significant quality decline (yellow color, off-odor) due to microbial spoilage. In summary, electron-beam treatment up to 3 kGy maintains the overall quality of fresh broccoli.     

Evaluation of the antioxidant potential of cauliflower (Brassica oleracea) from a glucosinolate content perspective

Glucosinolates (GLSs) are of great interest for their potential as antioxidant and anticancer agents. Our aim was to assess in vitro antioxidant activity of cauliflower GLSs and their derivatives, as well as to evaluate their contribution to the overall antioxidant capacity of cauliflower extracts using four different in vitro assays, 2,2′-azino-bis-(3-ethyl-benzothiazoline-6-sulphonic acid) radical cation (ABTS⁺), 2,2-diphenyl-1-picrylhydrazyl assay (DPPH), oxygen radical absorbance capacity assay (ORAC) and superoxide radical scavenging activity assay (SRSA). The antioxidant activities of GLSs determined using ABTS⁺ and DPPH assays were weak. In contrast, ORAC and SRSA assays showed significant antioxidant activity for GLSs. This activity was particularly important for glucobrassicin, which was the most effective GLS, and for glucoiberin and gluconapin. Mixtures of GLSs did not show either synergy or antagonism in general. Interestingly, the breakdown products, i.e. enzymatic hydrolysis-derived products (EHDP) of GLSs, were far more active than the native GLSs. The highest increases in antioxidant activity were obtained for EHDPs from progoitrin, sinigrin and glucoraphanin; progoitrin EHDP being the second most active derivative behind glucobrassicin EHDP. Antioxidant activities and GLS composition of different cauliflower by-product extracts were also determined. The contribution of different GLSs on measured antioxidant activity of cauliflower extracts was also discussed.     

Influence of pressure/temperature treatments on glucosinolate conversion in broccoli (Brassica oleraceae L. cv Italica) heads

Glucosinolates are a group of secondary plant metabolites that are found in the Brassicaceae family. Upon hydrolysis by the endogenous enzyme myrosinase, a large number of compounds can be formed of which some are potentially anticarcinogenic, while others are largely inactive. Furthermore, some bioactive compounds are unstable. Therefore, it is not only important to determine the type and amount of glucosinolates present or hydrolysed in a given plant, but it is also relevant to investigate the type of hydrolysis products. In this research, the effect of combined pressure–temperature treatments (100–500 MPa, 20–40 °C) on the glucosinolate conversion and the kind of hydrolysis products was studied in broccoli, both during treatment and after autolysis. The results, showed that high pressure can induce glucosinolate hydrolysis during treatment, promote the formation of isothiocyanates after treatment and that relatively more indole oligomers are formed during treatment than during autolysis. These results indicate that pressure treatment limits the loss of glucosinolates and its health beneficial products.     

Effect of hydrothermal treatment on the antioxidant properties of broccoli (Brassica oleracea var. botrytis italica) florets

In the extracts of fresh raw and frozen broccoli, caffeic, ferulic, sinapinic acids and kaempferol were identified. Boiling reduced the amounts of caffeic acid and kaempferol in both samples. The concentration of polyphenols was 2.69 mg/g fresh mass and 0.96 mg/fresh mass in fresh raw broccoli and frozen raw broccoli, respectively. Boiling significantly decreased the amounts of phenolic compounds in fresh broccoli (1.58 mg/g of fresh mass). In the case of frozen broccoli, boiling increased the concentration of polyphenols by 38%. Fresh broccoli extract neutralized free radicals by 19.87%. Boiling significantly reduced its antiradical activity (to 15.06%). Samples of frozen broccoli had a 27.06% antiradical ability. Boiling did not change the antiradical activity in frozen broccoli case. Hydrothermal processing significantly influenced on the ability of the extracts to inhibit the decolorization of β-carotene emulsion. The extract of fresh broccoli had a higher activity when uncooked. Boiling seemed to increase this activity in the case of frozen broccoli samples. The results of the studies on the ability of broccoli extracts to inhibit linoleic acid autooxidation were ambiguous and depended on the method applied. No correlation was found between the content of phenolic compounds and the antioxidant activity of the extracts, regardless of the experimental variant and technique used.     

Changes in carbohydrate and glucosinolate composition in white cabbage (Brassica oleracea var. capitata) during blanching and treatment with acetic acid

The effect of blanching and treatment with white vinegar containing acetic acid on dietary fibre, low-molecular-weight carbohydrates and glucosinolates was studied in two cultivars of white cabbage (Heckla and Predikant). The total content of dietary fibre and low-molecular-weight carbohydrates was similar in both cultivars (24 and 60 g/100 g DM, respectively), while the distribution between soluble and insoluble fibre differed (19% was soluble in Heckla versus 26% in Predikant, P < 0.01). Further, Heckla contained higher amounts of glucose and sucrose, while the content of fructose and total glucosinolates was lower than in Predikant. The content of individual glucosinolates differed between the two cultivars. During blanching there was a loss of dry substance (30–34 g/100 g DM), where low-molecular-weight carbohydrates primarily explained the loss (82–90%), but some of the loss was also dietary fibre (about 8%), both soluble fibre containing uronic acids (mainly Predikant) and insoluble ones containing glucose (mainly Heckla). The glucosinolate levels decreased substantially in both cultivars, although the total loss was higher in Predikant (74%) than in Heckla (50%). The individual glucosinolates were affected to different degrees (15–91%). During souring with acetic acid, the content of dietary fibre (primarily insoluble ones) decreased further, while the content of low-molecular-weight carbohydrates was less affected. The total content of glucosinolates was not affected in Heckla but was further reduced in Predikant. There was, however, a substantial increase in 4-methoxyglucobrassicin in both cultivars. It is concluded that blanching and souring decrease the content of carbohydrates and glucosinolates to a great extent and both cultivars behaved similarly. However, individual components were affected differently in the two cultivars.     

Characterization of polyphenol oxidase from broccoli (Brassica oleracea var. botrytis italica) florets

Polyphenol oxidase (PPO) from broccoli florets was extracted and purified through (NH₄)₂SO₄ precipitation, ion-exchange and gel filtration chromatography. The molecular weight was estimated to lie between 51.3 and 57 kDa by sodium dodecyl sulphate-polyacrylamide gel electophoresis (SDS-PAGE) and gel filtration. The effects of substrate specificity, pH, and sensitivity to various inhibitors: citric acid, ascorbic acid, sodium sulphate and EDTA (sodium salt of ethylenediaminetetraacetic acid) of partially purified PPO were investigated. Polyphenol oxidase showed the best activity toward catechol (K_M = 12.34 ± 0.057 mM, V_max = 2000 ± 8736 U/ml/min) and 4-methyl catechol (K_M = 21 ± 0.087 mM, V_max = 28.20 ± 0.525 U/ml/min). The optimum pH for broccoli PPO was 5.7 with catechol and 4-methylcatechol as substrates. The most effective inhibitor was sodium sulphate.     

The effect of temperature and radiation on flavonol aglycones and flavonol glycosides of kale (Brassica oleracea var. sabellica)

The winter crop kale has a complex profile of different glycosylated and acylated flavonol glycosides which may be affected by global warming. To the best of our knowledge, compound–climate relationships for flavonol aglycones and flavonol glycosides were established for the first time. The investigated 10 major flavonol glycosides responded structure-dependent in the investigated temperature range between 0 and 12 °C and the photosynthetic active radiation range between 4 and 20 mol m⁻² d⁻¹, e.g. the decrease in temperature led to an increase in sinapic acid monoacylated and diacylated quercetin glycosides, while the sinapic acid monoacylated kaempferol glycosides showed a maximum at 4.5 °C. Furthermore, the hydroxycinnamic acid residues and the different number of glucose moieties in the 7-O position affected the response of kaempferol triglucosides. Consequently, global warming would result in lower concentrations of antioxidant-relevant quercetin glycosides in winter crops, suggesting a production at e.g. higher altitudes due to lower temperature.     

Influence of combined hot air and UV-C treatment on the antioxidant system of minimally processed broccoli (Brassica oleracea L. var. Italica)

Broccoli develops a fast senescence that deteriorates the nutritional quality of the product. In this work, a combined treatment with hot air and UV-C were applied to minimally processed broccoli florets to investigate their effects on the antioxidant system during storage at 20 °C. Application of UV-C and heat enhanced by approximately 13% the levels of total antioxidants immediately after treatment. These levels were lower than initials in both control and treated samples. Nevertheless, higher values (ranged from 12 to 50%) were also detected in treated samples during storage. In general, higher values of antioxidants were correlated with elevated levels of phenols (from 10 to 17%) and ascorbic acid (from 11 to 17%) in treated florets. The increment in the content of phenolics in treated broccolis was correlated to an increased phenylalanine ammonia lyase (PAL) activity. The combined treatment also affects the activity of enzymes involved in removal of reactive oxygen species. An increase of about 50% in superoxide dismutase activity was detected after treatment. During the first days of storage this higher activity was about 40%. Enzymes that remove H₂O₂ like catalase and ascorbate peroxidase showed an enhanced activity toward the end of storage. The combined treatment diminished the peroxidase (POX) activity approximately 60% after the treatment and near 50% after two days of storage, suggesting a minor role of this enzyme in detoxification of H₂O₂. In conclusion, combined treatment may contribute to enhance the protection against oxidative molecules not only by increasing levels of phenolics and ascorbic acid but also by enhancing the activity of enzymes involved in removing reactive oxygen species.     

Genotypic and climatic influences on the concentration and composition of flavonoids in kale (Brassica oleracea var. sabellica)

The aim of this study was to determine the composition and concentration of flavonoid aglycones in kale, the dependence on genotype and their interaction with decreasing temperature and global radiation. Eight kale cultivars, comprising hybrid and traditional, old cultivars, were grown in a field experiment and harvested four times at 4-week intervals. The traditional, old cultivars in particular contained high concentrations of flavonoids. In all of the investigated cultivars, kaempferol was the main flavonoid aglycone, followed by quercetin and isorhamnetin, which was quantified in six of the eight cultivars. Furthermore, in six of the eight cultivars, the total concentration of flavonoids remained unchanged with decreasing temperature and global radiation. The quercetin concentration increased in five of these six cultivars, whereas the kaempferol concentration decreased. Interestingly, the quercetin-to-kaempferol ratio increased in all of the investigated cultivars, despite the fact that the radiation level decreased, suggesting that the impact of the decline in temperature could be responsible for this effect.     

Simultaneous evaluation of intact glucosinolates and phenolic compounds by UPLC-DAD-MS/MS in Brassica oleracea L. var. botrytis

A method for the simultaneous determination of intact glucosinolates and main phenolic compounds (flavonoids and sinapic acid derivatives) in Brassica oleracea L. var. botrytis was proposed. A simplified sample extraction procedure and a UPLC separation were carried out to reduce the total time of analysis. B. oleracea samples were added with internal standards (glucotropaeolin and rutin), and extracted with boiling methanol. Crude extracts were evaporated under nitrogen, redissolved in mobile phase and analysed by UPLC with double detection (ESI⁻-MRM for glucosinolates and flavonoids, and DAD for main sinapic acid derivatives). The proposed method allowed a satisfactory quantification of main native sinapic acid derivatives, flavonoids and glucosinolates with a reduced time of analysis.     

Partial characterization of white cabbages (Brassica oleracea var. capitata f. alba) from different regions by glucosinolates, bioactive compounds, total antioxidant activities and proteins

Glucosinolates (GLS), antioxidative compounds, total radical scavenging activities (TRSAs) and proteins of white cabbage samples derived from different regions of Europe, collected in the spring and autumn, were studied. Glucobrassicin and sinigrin were the dominating GLS in all analyzed cabbage samples. Depending on origin, these two GLS accounted for ∼30% to ∼70% of the total. The total GLS content ranged from 3.3 to 7.7 μmol/g dw in lyophilized vegetables. Assays based on electron transfer [total phenols by Folin-Ciocalteu reagent (FCR), 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] radical cation (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH)] were used to compare the TRSAs and the main bioactive compounds in cabbage. Total polyphenols varied from 2.4 to 4.9 GAE/g dw. The TRSAs ranged from 2.7 to 8.2 μmol TE/g dw in ABTS test and from 2.4 to 5.4 μmol TE/g dw in DPPH assay. The maximum amount of polyphenol compounds, antioxidant activity, as well as total GLS content, were recorded in Belgian cabbage harvested in the autumn and the lowest ones were found for Poland 2 cabbage harvested in the spring. In extracted and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) proteins from cabbage leaves only in samples from England and Belgium some differences in patterns were found in the regions of 60 and 97 kDa. The calculated correlations between antioxidative potency and the abundance of bioactive compounds were highly statistically significant. This suggests that TRSA could serve as a means of standardization of natural mixtures, at least in the case of cabbage, necessary to compare results of biological studies carried out for vegetable derived samples.     

Effect of thermal treatment on glucosinolates and antioxidant-related parameters in red cabbage (Brassica oleracea L. ssp. capitata f. rubra)

The effects of various thermal processing treatments (blanching, boiling and steaming) of red cabbage, Brassica oleracea L. ssp. capitata f. rubra cv. ‘Autoro’, were assessed for the levels of glucosinolates (GLS), total phenols (TP), total monomeric anthocyanins (TMA), l-ascorbic acid (L-AA) and soluble sugars, as well as for the antioxidant potential by the ferric reducing ability power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Individual native GLS were determined by ion-pair HPLC-MS/DAD. There were significant (p < 0.05) losses in blanched red cabbage: TP, 43%, TMA 59%, FRAP 42%, ORAC 51%, L-AA 48% and soluble sugars 45%. Boiling gave less extensive reductions: TP 16%, TMA 41%, FRAP 17%, ORAC 19%, L-AA 24% and soluble sugars 19%. Steaming caused no losses for TP, ORAC, FRAP or soluble sugars. However, significant reductions were found for TMA and L-AA, with 29% and 11%, respectively. In general losses were accounted for in the processing waters; however, TMA was not fully recovered, indicating degradation. Total GLS were severely affected by processing, with reductions of 64%, 38% and 19% in blanched, boiled and steamed red cabbage, respectively. Total aliphatic and indole GLS were similarly affected. Lost GLS were partially recovered in the processing water.     

Impact of UV-B irradiation on chlorophyll degradation and chlorophyll-degrading enzyme activities in stored broccoli (Brassica oleracea L. Italica Group) florets

UV-B irradiation was applied to broccoli florets to investigate its effect on chlorophyll degradation and chlorophyll-degrading enzyme activities in stored broccoli. Broccoli florets were irradiated with UV-B doses at 4.4, 8.8, and 13.1 kJ m⁻² and then kept at 15 °C in darkness. We found that a UV-B dose of at least 8.8 kJ m⁻² efficiently delayed the decrease of the hue angle value and the contents of chlorophylls a and b. Chlorophyllide a and 13²-hydroxychlorohyll a gradually decreased with senescence. Pheophorbide a and pyropheophorbide a levels were significantly higher in broccoli without UV-B treatment. Chlorophyllase and chlorophyll-degrading peroxidase activities with UV-B treatment were suppressed, as well as the activity of Mg-dechelatase. Mg-dechelating substance activity was also suppressed with this treatment. We concluded that UV-B treatment effectively suppressed chlorophyll degradation in broccoli florets during storage, suggesting that the effect could be due to the suppression of chlorophyll-degrading enzyme activities.     

Effect of preliminary processing and method of preservation on the content of selected antioxidative compounds in kale (Brassica oleracea L. var. acephala) leaves

Vitamin C and polyphenol content as well as total antioxidative activity were investigated in fresh leaves of kale; in leaves after blanching or cooking; and in frozen and canned leaves. In 100 g fresh matter, kale leaves contained 384.9 mg polyphenols and 112.1 mg vitamin C, with a Trolox equivalent antioxidant capacity (TEAC) level of 1175 μM Trolox. Of the polyphenols identified in kale leaves, ferulic acid occurred in the highest amount (240.44 mg/100 g, constituting 62% of total polyphenols). Freezing was a better method of preserving kale leaves since the loss of nutritive constituents was lower than in the case of canning. Depending on preliminary processing and storage temperature, after one-year storage frozen leaves contained 82.9–171.3 mg polyphenols and 39.3–65.4 mg vitamin C, with TEAC at the level of 501–681 μM Trolox in 100 g. In canned leaves these values were: 91.3–94.1 mg polyphenols, 16.1–19.3 mg vitamin C and 268–293 μM Trolox.     

Anthocyanin composition of cauliflower (Brassica oleracea L. var. botrytis) and cabbage (B. oleracea L. var. capitata) and its stability in relation to thermal treatments

Violet cauliflower and red cabbage were analysed for their anthocyanin profiles before and after thermal treatments. Anthocyanins are well-noted as healthy compounds due to their antioxidant properties. Samples were analysed for total anthocyanin content by using a spectrophotometric differential pH method. An MS-based method, combining high-performance liquid chromatography (HPLC) with quadrupole tandem mass spectrometry (HPLC–MS/MS) was developed, aimed to separate, identify and quantify the main anthocyanin forms. The procedure involves a rapid and efficient pre-treatment of the samples by solid-phase extraction, followed by selective determination of all compounds in a single run analysis using HPLC–MS/MS. Structural information for the identification of compounds was obtained from their fragmentation patterns (MS/MS spectra). The compounds were separated by HPLC and detected in the multiple reaction monitoring mode (MRM), which provides a high level of selectivity for targeting the analytes in vegetables. Cauliflower and red cabbage showed differences in their anthocyanin profiles: cyanidin-3,5-diglucoside was absent in cauliflower, while it was well represented in red cabbage, together with the characteristic anthocyanin of Brassica genus, cyanidin-3-sophoroside-5-glucoside. The p-coumaryl and feruloyl esterified forms of cyanidin-3-sophoroside-5-glucoside were predominant in cauliflower, while the sinapyl one was mostly present in red cabbage. Besides, the stability of cauliflower’s anthocyanin profile was evaluated in relation to thermal pre-treatments. All thermal treatments, except microwave heating, drastically reduced total cauliflower anthocyanin content. The amount of individual anthocyanins was expressed as the percentage with respect to total anthocyanin amount, spectrophotometrically measured. Significant individual changes were observed after different thermal treatment with an isomer formation.     

Steam processed broccoli (Brassica oleracea) has higher antioxidant activity in chemical and cellular assay systems

Thermally processed fruits and vegetables have long been considered to have a lower nutritional value compared with the fresh produce. This consideration is based on the fact that vitamin C or other thermolabile compounds may lose their activity due to oxidation or in consequences of leaching into the water during home cooking or industrial processing, such as blanching. In this study, broccoli, one of the major agricultural products was exposed to steam; and total antioxidant activity was measured by oxygen radical capacity (ORAC) analysis and a cellular measure of reactive oxygen species (ROS) assay. The study also evaluated the level of total phenolics and total flavonoid content in fresh and steam processed broccoli samples. The result clearly showed that steam-processing elevated the total ORAC (hydrophilic, lipophilic) value by 2.3 fold. Measuring cellular levels of ROS, we found that the hydrophilic part of a steam processed broccoli had a significant reduction of 2,2′-azobis [2-amidinopropane] dihydrochloride (AAPH) induced intracellular ROS level in comparison to that of fresh counterpart. Total phenolic content and total flavonoid content also increased in steamed-processed broccoli. Chromatographic analysis showed that a non-phenolic fraction which appeared unbound on a cosmosil 140 C18 resin contributed 54%, and the phenolic pool eluted by 50–80% methanol contributed 41% of its total increase in ORAC value. Steam processed broccoli gave significant cytoprotection in PC-12 cell line and this neuroprotective efficacy warrant further investigation. This information may have a significant impact on consumers’ food selection, depending on the processing technology used to process agricultural products.     

Effect of preliminary processing, method of drying and storage temperature on the level of antioxidants in kale (Brassica oleracea L. var. acephala) leaves

The levels of vitamin C, polyphenol constituents and Trolox equivalent antioxidant activity (TEAC) were analyzed in raw kale leaves, blanched leaves and dried leaves obtained using air and freeze-drying methods. 100 g of raw kale leaves contained 683 mg vitamin C and 2236 mg polyphenols (identified using the HPLC method); the level of antioxidative activity was 71 μM Trolox/1 g dry matter. Compared with the raw material, blanching before drying brought about significant decreases of 15% in vitamin C, 32% in polyphenols and13% in TEAC. After 12-month storage, air-dried material retained 30-37% polyphenols; 43-57% vitamin C; and 41-50% of the initial TEAC level; the corresponding values for freeze-dried material were 40-47%; 50-65% and 54-66% depending on the type of sample. Freeze-dried kale leaves contained higher levels of antioxidants than air-dried material: polyphenols, vitamin C and TEAC were respectively 36%, 15%and 33% higher.