Processing (blanching, boiling, steaming) effects on the content of glucosinolates and antioxidant-related parameters in cauliflower (Brassica oleracea L. ssp. botrytis)

Five different varieties of cauliflower (Brassica oleracea L. ssp. botrytis); two white (cv. ‘Aviso’, ‘Dania’), one purple (cv. ‘Grafitti’), one green (cv. ‘Emeraude’) and one romanesco/green pyramidal (cv. ‘Celio’) cultivar have been studied. All samples were thermally processed and the effects on the levels of glucosinolates (GLS), total phenols (TP), total monomeric anthocyanins (TMA), l-ascorbic acid (l-AA) and antioxidant capacities (FRAP and ORAC) were investigated. Processing methods applied were: blanching (3 min), boiling (10 min) and steaming (10 min). Total GLS were significantly (p < 0.05) affected by processing with the highest losses, 55 and 42% on average, occurring for boiled and blanched samples, respectively. Significant effects were also noted for steaming, but to a lesser extent, i.e. 19% average reduction. Antioxidant-related parameters were similarly affected with average losses of 27, 33, 36 and 46% in boiled cauliflower and 16, 21, 22 and 28% in blanched for TP, FRAP, l-AA and ORAC, respectively. Blanching and boiling reduced TMA in purple cauliflower by 38 and 53%, respectively. Steaming affected the antioxidant-related parameters the least for all cultivars. l-AA was significantly reduced by 14% in all cultivars by steaming. Some differences in behaviour between cultivars were noted, especially between white and coloured cultivars for TP, FRAP and l-AA, but also for some GLS. The main losses were caused by leaching into the processing water.     

Glucosinolates, l-ascorbic acid,total phenols,anthocyanins, antioxidant capacities and colour in cauliflower (Brassica oleracea L. ssp. botrytis); effects of long-term freezer storage

Blanching and one year freezer storage were performed on one purple, two white and two green varieties of cauliflower (Brassica oleracea L. ssp. botrytis) to assess the effects on several health-related phytochemicals. Blanching, prior to freezing, reduced total aliphatic and indole glucosinolates (GLS) by 31% and 37%, respectively. l-ascorbic acid (L-AA), total phenols (TP), anthocyanins, FRAP and ORAC were on average reduced by 19, 15, 38, 16 and 28%, respectively. The colour measured as CIELAB parameters was partly affected by blanching: lightness was significantly reduced in the white cultivars and in a romanesco type cultivar. Chromaticity was reduced in the coloured cultivars with a twice-as-large reduction in the purple cultivars. Hue increased for all samples, except for the purple cultivar where a decrease was found. Long-term freezer storage did not affect total aliphatic and indole GLS in cauliflower in a major way. Freezer storage did result in an average L-AA decrease of 24% for all but the purple cultivar. Some reductions in the TP levels were also found but not to the extent found for L-AA. When an effect was found in the FRAP and ORAC values, the reductions occurred towards the end of the storage period and were 15% and 37%, on average, respectively. Long-term freezer storage did not affect the anthocyanin content and only minor effects were found for the colour parameters.     

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.     

Variation of glucosinolates in 62 varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) and their antioxidant activity

Glucosinolate (GSL) and antioxidant activity in 62 varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) were determined by HPLC and DPPH, HRSA, and FRAP assays. Five aliphatic GSLs: progoitrin, sinigrin, glucoalyssin, gluconapin, and glucobrassicanapin; four indolyl GSLs: 4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin; one aromatic GSL: gluconasturtiin were identified. Glucobrassicanapin and gluconapin documented the most abundant (average 4.52 and 3.72 μmol/g DW, respectively). The contents of total GSLs varied extensively among 62 varieties (range from 2.83 to 48.53 μmol/g DW). Comprehensive differences in total and individual GSL contents have also been observed among different varieties. Indolyl and aromatic GSL together accounted 26% of the total GSLs; but there are few differences among varieties. FC7 and FI17 could be good candidates for future breeding programs since they had a high quantity of glucobrassicin (2.10 and 1.66 μmol/g DW, respectively). Most of the Chinese cabbage varieties showed significant antioxidant activities when compare with positive control. However, three antioxidant assays were not significantly correlated with total GSLs. The presence of significant quantities of glucobrassicin in some varieties should be studied more extensively, since GSL is the precursor of indole-3-carbinol, a potent anticancer isothiocyanate.     

Analysis and metabolite profiling of glucosinolates,anthocyanins and free amino acids in inbred lines of green and red cabbage (Brassica oleracea L.)

We profiled and quantified glucosinolates (GSLs), anthocyanins and free amino acids in thirty-seven inbred lines green and red cabbage. Analysis of these distinct cabbages revealed the presence of 8 GSLs, 13 anthocyanins and 12 free amino acids. GSL contents were varied among the different lines of cabbage. The maximum levels of glucoraphanin (14.91 μmol/g DW) and glucobrassicin (12.37) were found in FX 1-28 and FX 1-32 lines, respectively. Total GSLs in red cabbage lines were 50% higher than those of green cabbage. Anthocyanin contents in red cabbage were ranged from 4.11 to 6.81 mg/g DW in FX 2-3 and FX 1-34 lines, respectively. Among the 13 anthocyanins, both cyanidin 3-(feruloyl) (sinapoyl)diglucoside-5-glucoside and cyanidin 3-(sinapoyl) (sinapoyl)diglucoside-5-glucoside levels were the highest amounts. The amounts of total free amino acids were ranged from 523.5 to 1308 mg/100 g fresh weight (FW) in green cabbage and 484.8 to 1271 in red cabbage, respectively. In red cabbage lines, 9.4% of the total free amino acids accounted essential amino acids such as valine, threonine, isoleucine, leucine and lysine. Thus, the amounts of GSLs, anthocyanins, and amino acids varied widely, and the variations in these compounds between the lines of cabbage were significant.     

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.     

Characterisation of the pigment components in red cabbage (Brassica oleracea L. var.) juice and their anti-inflammatory effects on LPS-stimulated murine splenocytes

To determine anti-inflammatory effects of pigments from red cabbage, red cabbage (Brassica oleracea L. var.) juice was prepared, characterized by UV–vis absorption spectra, partially purified by Sephadex LH-20 column, analyzed by HPLC, and administered to lipopolysaccharide (LPS)-stimulated murine splenocyte cultures. The study showed that red cabbage juice (RC) exhibited anti-inflammatory effects against LPS-induced inflammation of splenocytes via increasing anti-inflammatory cytokine interleukin (IL)-10 and decreasing pro-inflammatory cytokine IL-6 secretions. The maximum absorption peaks of RC and its heated products, but not activated charcoal-adsorbed products, appeared at 280 nm with a small shoulder around 310–330 nm while there existed a minor peak at 560 nm (range from 480 to 630 nm), reflecting red cabbage juice included phenolics, flavonoids, and anthocyanins. The lyophilized powder of chromatographic fractions F2, F3, and F4 through Sephadex LH-20 column were rich in phenolics (5.9 ± 0.2%, 4.4 ± 0.0%, and 3.9 ± 0.0%, respectively) and flavonoids (1.8 ± 0.3%, 1.8 ± 0.3%, and 1.1 ± 0.3%, respectively). The results suggest that anti-inflammatory pigment compounds in red cabbage juice were heat stable. Further analysis of chromatograms from HPLC suggests malvidin glycosides including malvidin 3-glucoside (oenin), malvidin 5-glucoside and malvidin 3,5-diglucoside in red cabbage juice could inhibit IL-6 secretion of LPS-stimulated splenocytes.     

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.     

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.     

Effect of water content and temperature on glucosinolate degradation kinetics in broccoli (Brassica oleracea var. italica)

The effect of water content and temperature on glucosinolate thermal degradation in broccoli (Brassica oleracea var. italica) was investigated. Broccoli was freeze dried obtaining batches with water content between 13% and 82% (a_w from 0.32 to 0.99). These samples were heated at different temperatures (from 60 to 120 °C) and glucosinolate levels were monitored. To rule out enzymatic breakdown, myrosinase was inactivated prior to heating. Degradation could be described by first-order kinetics for all glucosinolates and all water contents. In the temperature range 60–100 °C the sample with 13% showed the lowest degradation rate, whereas at 120 °C the degradation rate increased with the water content. This particular behavior was reflected by the high activation energy value of the driest sample. Several hypotheses to explain the observed behavior are discussed.     

Application of thermal inactivation of enzymes during vitamin C analysis to study the influence of acidification,crushing and blanching on vitamin C stability in Broccoli (Brassica oleracea L var. italica)

The effectiveness of heat inactivation of oxidative enzymes e.g., ascorbic acid oxidase (AAO) to stabilise vitamin C during extraction and analysis was evaluated. The influence of different sequences of performing treatments including acidification (pH 4.3 vs. pH 6.5), crushing, high temperature short time (90 °C/4 min–HTST) and low temperature long time (60 °C/40 min–LTLT)) blanching on vitamin C stability in broccoli florets and stalks was also investigated. Heat inactivation of enzymes prior to matrix disruption resulted in higher vitamin C values mainly in L-ascorbic acid (L-AA) form, while lack of enzyme inactivation resulted in high vitamin C losses resulting from conversion of L-AA to dehydroascorbic acid. Various treatments and their sequence of application influenced vitamin C stability as follows: (i) crushing prior to blanching reduced vitamin C stability and (ii) in the absence of heating, acidification increased vitamin C stability (iii) blanching prior to crushing resulted in higher vitamin C retention, with HTST blanching retaining more vitamin C than LTLT blanching.     

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.     

Antioxidant capacity,total phenolic and ascorbate content as a function of the genetic diversity of leek (Allium ampeloprasum var. porrum)

Extracts of the white shaft and green leaves of 30 leek cultivars were investigated for their antioxidant properties, total phenolic (TP) and l-ascorbic acid (AA) content. The measured antioxidant properties included free radical scavenging activities against peroxyl (ORAC) and 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH) and their Fe³⁺ reducing capacity (FRAP). The results from this study suggest that the green leek leaves generally have significantly stronger antioxidant properties than the white shaft. Correlation analysis between the TP and the AA content and the antioxidant activity showed that phenolics and ascorbic acid contribute significantly to the antioxidant activity of leek. The three antioxidant activity assays were all correlated for the extracts of the white shaft of the 30 leek cultivars. Principal component analysis (PCA) elucidated the influence of part and type of cultivar on the antioxidant capacity, TP, and l-ascorbic acid content, whilst the breeding strategy and seed company had no influence.     

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.     

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.     

Influence of steaming treatment on chemical and technological characteristics of einkorn (Triticum monococcum L. ssp. monococcum) wholemeal flour

Einkorn (Triticum monococcum ssp. monococcum) is an underutilised wheat with high protein, lutein and tocols content, particularly suited for baby and specialty foods. To study the influence of kernel steaming treatments on chemical and technological properties, seeds of five einkorn accessions and one bread wheat control (cv. Blasco) underwent different steaming conditions, and their wholemeal flours were compared for ash, protein, lutein, tocols and α-amylase content, SDS sedimentation volume and gelatinisation parameters. Furosine, a heat damage marker, was measured as well. The treatments significantly influenced most of the parameters. In particular, lutein, tocols and α-amylase diminished after steaming; SDS sedimentation volumes and most gelatinisation parameters also decreased, whereas gelatinisation temperatures and furosine contents increased. The changes were stronger under more drastic steaming conditions, although treatment × genotype influence was sometimes detected. Steaming induced migration of lutein and tocopherols from the bran and germ fractions to the kernel endosperm.     

Microwave inactivation of red beet (Beta vulgaris L. var. conditiva) peroxidase and polyphenoloxidase and the effect of radiation on vegetable tissue quality

The inactivation of polyphenoloxidase (PPO) and peroxidase (POX) in red beet by traditional and microwave (MW) blanching was studied. Microwave heating effects on color and texture were also studied.Red beet subjected to MW blanching for 5 min at 100-200 W resulted in large weight losses accompanied by a high degree of shrinking. POX was the more heat resistant enzyme. The 90% destruction (D value) of the activity of both enzymes could be achieved only at 200 W within the 5 min frame employed for the tests.When the samples were immersed in water and both the food sample and the water were submitted to MW exposure at 250-450 W or variable power with a maximum at 935 W, shrinking was avoided. The D value at 90 °C (reference; D_Tref) and z could be determined after time-temperature corrections, and it was observed that, in general, D_Tref values for POX were smaller than for PPO. The microwave treatment (maximum power: 935 W) designed to provide a similar temperature profile to the one observed for traditional blanching (immersion in water at 90 °C), showed the smallest D_Tref value for POX inactivation. All treatments reduced elastic characteristics and changed the color of the tissues showing a shift to blue mainly in the case of microwave processes.