Microwave-assisted extraction of betalains from red beet (Beta vulgaris)

The use of microwave assisted extraction (MAE) was investigated in this work for the extraction of betalains from diced red beets. Several treatments with different combinations of time, power and duty cycle applied to the samples were studied. The combination of 400 W and 100% duty cycle for 90–120 s resulted in the highest amount of recovered betanines; whereas at 140–150 s the highest amount of betaxanthins was obtained. The addition of ascorbic acid (0.040 mol/L) to the extracting solvent and the development of a two-step MAE process with a cooling period in-between and after processing steps led to an enhancement in the amount of pigments obtained. The effect of extraction time at each extraction step on betalains yield was determined by applying a factorial design and surface plots were constructed. The duration of the second step significantly affected the yield of betanines and betaxanthins obtained (p < 0.05). A prediction model was proposed and validated to meet the optimal extraction times. Betalain yields obtained by MAE were twice as high as those obtained during conventional extraction and conventional extraction at 80 °C.     

Stability of yellow-orange cactus pear (<Emphasis Type="Italic">Opuntia ficus-indica</Emphasis> [L.] Mill. cv. ‘Gialla’) betalains as affected by the juice matrix and selected food additives

The impact of added ascorbic, isoascorbic, and citric acid on heat stability of cactus pear betalains was investigated at pH 4 and 6, respectively. To assess the influence of cactus pear juice matrix compounds such as sugars, organic acids, amino acids, and pectic substances on pigment stability, juice and purified pigment preparations derived therefrom were assayed separately. Immediately after heating at 85 °C for 1 h and a 24 h cool storage period, betalain retention, betanin and proline-betaxanthin isomerisation as well as overall colour changes were determined. Pigment stability and colour characteristics depended on type and concentration of the respective additive as well as on pH conditions. Interestingly, the stabilising effects of the additives under investigation were less pronounced in matrix-free pigment preparations compared to juice samples. Maximum pigment retention amounting to 79% was obtained when 0.1% citric acid was added to the juice at pH 6. For the first time, formation of 2-decarboxy-betanin upon thermal treatment of yellow-orange cactus pear juice being more distinct at pH 4 as compared to pH 6 is reported.     

Effects of maturity stage and storage on cactus berry (Myrtillocactus geometrizans) phenolics,vitamin C,betalains and their antioxidant properties

The CIE-LAB coordinates, total phenols, betalains and vitamin C were determined in berry cactus fruit at different maturity stages, before and after storage. The contribution of those antioxidant compounds to the Trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) were assessed as well. The simple phenolics profile was determined by high performance liquid chromatography with diode array detection (HPLC-DAD). After storage, the fruit develops similar colour compared to ripe and overripe fresh fruit. Total phenols and vitamin C were reduced by up to 82.4% and 68.5%, respectively, as fruit matures and after storage. On the contrary, beta-cyanins increase up to 51.8% as fruit matures and after storage. Betalains contribute with 58.4% to TEAC and up to 43.4 to ORAC values. Gallic acid and quercetin were the major simple phenolics present. Due to the contents of antioxidant compounds and the outstanding antioxidant activities, cactus berries could contribute to health.     

Characterisation and determination of in vitro antioxidant potential of betalains from Talinum triangulare (Jacq.) Willd.

Talinum triangulare (Jacq.) Willd is an erect, succulent, perennial herb belonging to the family Portulacaceae. Under extreme environmental conditions, the plant produces betalain pigments which get accumulated in the stem region. Hence, in the present study, the betaxanthin and betacyanin patterns from different samples of T. triangulare have been investigated by applying high-performance liquid chromatography photo-diode array detection (HPLC-PDA) coupled with positive ion electro-spray mass spectrometry. Two betacyanins and two betaxanthins were identified in aqueous methanolic extract of flower, stem and leaf. Betanin, isobetanin, immonium conjugates of betalamic acid with dopamine and tyrosine were elucidated. The total betalain content was estimated by photometric analysis. In vitro antioxidant activity for the betalain extract determined by various methods revealed potent scavenging ability. The current work may possibly be considered beneficial in utilisation of the plant T. triangulare as a natural colourant in food and beverage industries.     

Juice matrix composition and ascorbic acid fortification effects on the phytochemical,antioxidant and pigment stability of açai (Euterpe oleracea Mart.)

The effects of juice matrix composition on the phytochemical stability of açai (Euterpe oleracea Mart.) were evaluated by contrasting natural clarified juice systems with isolated polyphenolic and anthocyanin fractions, in the presence or absence of ascorbic acid (500 mg/l) under accelerated storage conditions (37 °C). Polyphenolic (anthocyanin and non-anthocyanin polyphenolics) and anthocyanin fractions were isolated using C18 Sep-Pak columns and then re-dissolved in the original aqueous juice matrix (unbound fraction) or in a citric acid buffer (pH 3.5). The isolation of anthocyanins from the açai matrix improved their colour stability but a greater retention of non-anthocyanin polyphenolics and antioxidant properties was favoured by the initial juice composition. The presence of non-anthocyanin polyphenolics exerted a protective effect against ascorbic acid oxidation and enhanced polyphenolic and antioxidant stability in isolates fortified with ascorbic acid. However, all isolates obtained from açai experienced significant colour, polyphenolic, and antioxidant losses during storage and this indicated that optimization of early stages of industrial processing, storage and distribution are necessary to retain the functional properties of açai-containing products.     

Acid tolerant mutants of Bifidobacterium animalis subsp. lactis with improved stability in fruit juice

The potential of UV mutagenesis combined with a specific selection step to generate more acid-resistant Bifidobacterium animalis subsp. lactis Bb-12 strains with improved viability in low pH food matrices was investigated in this study. A total of 144 Bb-12 UV-mutants were initially characterised. After prolonged storage in apple juice (pH 3.5) and various pheno- and genotypic tests (acid and bile tolerance, substrate utilisation, antibiotic susceptibility, aerotolerance, RAPD) two mutants (2.20 and 2.56) were chosen for further studies including cell surface morphology, stability of various traits after repeated inoculations, and performance in a fermenter and during down-stream processing. 2.20 and 2.56 showed over two Log-values better viability in pH 3.5 juice compared to Bb-12. Alterations in cell surface structures of 2.20 and 2.56 were detected with AFM, whereas other studied traits remained unchanged.In conclusion, UV mutagenesis and subsequent incubation in acidic medium enabled improving the stability of B. animalis subsp. lactis in low pH juice. Acid tolerance testing (HCl, pH 2.5, 2 h) results did not predict long-term stability of the strains in acidic food matrix (apple juice pH 3.5).     

Screening of antioxidative properties of the methanolic extracts of Pelargonium endlicherianum Fenzl., Verbascum wiedemannianum Fisch. & Mey., Sideritis libanotica Labill. subsp. linearis (Bentham) Borm., Centaurea mucronifera DC. and Hieracium cappadocicum Freyn from Turkish flora

Five extracts from five plants, of which four are endemic to Turkish flora, were screened for their possible in vitro antioxidant activities by two complementary test systems, namely DPPH free radical-scavenging and β-carotene/linoleic acid. In the first case, Pelargonium endlicherianum extract exerted greater antioxidant activity than those of other plant extracts studied with an IC₅₀ value of 7.43 ± 0.47 μg/ml, followed by Hieracium cappadocicum (30.0 ± 0.14 μg/ml). When compared to the synthetic antioxidant BHT (18.0 ± 0.40 μg/ml), the methanolic extract of P. endlicherianum exhibited more than two fold greater antioxidant activity. In the β-carotene/linoleic acid test system, the most active plant was P. endlicherianum with 72.6% ± 2.96 inhibition rate, followed by H. cappadocicum (55.1% ± 2.33) and Verbascum wiedemannianum (52.5% ± 3.11). Antioxidant activities of curcumin and ascorbic acid were also determined as positive controls in parallel experiments.     

Comparative chemical composition,antioxidant and hypoglycaemic activities of Juniperus oxycedrus ssp. oxycedrus L. berry and wood oils from Lebanon

Juniper (Juniperus oxycedrus) is used in European cuisine for its distinguishing flavour. J. oxycedrus ssp. oxycedrus berry and wood essential oils were tentatively identified by GC and GC/MS. Fifty compounds were identified in the berry oil and 23 compounds were identified in the wood oil. The J. oxycedrus ssp. oxycedrus berry oil was characterised by high contents of α-pinene (27.4%) and β-myrcene (18.9%). Other important compounds were α-phellandrene (7.1%), limonene (6.7%), epi-bicyclosesquiphellandrene (2.3%) and δ-cadinene (2.2%) while, in the wood oil, δ-cadinene (14.5%) is a major main component, together with cis-thujopsene (9.2%) and α-muurolene (4.9%). In vitro evaluation of antioxidant activity by the DPPH method showed a significant activity for both oils with IC₅₀ values of 1.45 μl/ml for wood and 7.42 μl/ml for berries. Hypoglycaemic activity was investigated through the inhibition of α-amylase. The results revealed that oil obtained by hydrodistillation from J. oxycedrus ssp. oxycedrus wood exhibits an interesting activity with IC₅₀ of 3.49 μl/ml.     

Nutritional components and anti-oxidant capacity of ten cultivars and lines of cactus pear fruit (Opuntia spp.)

Cactus pear fruit (Opuntia) are harvested from various species of the genus Opuntia of the cactus family (Cactaceae), and are produced and consumed in several countries. We have characterized the nutritional content and antioxidant capacity (AC) of the fruit of ten cultivars/lines of distinct pulp colors. ‘Camuesa’ had the highest betalains, total carotenoids, ß-carotene, ascorbic acid, and was one of the highest in total phenolic compounds, but its AC did not demonstrate outstanding differences with some other cultivars/lines that were not as rich in these compounds. ‘Roja Pelota’ had high AC when measured with the DPPH assay, but had low total carotenoids, ß-carotene and total phenolic content. ‘Reyna’ had slightly low AC as measured by the FRAP assay, practically no betalains, and low vitamin C, tocopherols, ß-carotene, and total phenolic content.     

Effect of endogenous ascorbic acid oxidase activity and stability on vitamin C in carrots (Daucus carota subsp. sativus) during thermal treatment

The purpose of this research was to study the effect of endogenous ascorbic acid oxidase (AAO) on vitamin C in carrots (Daucus carota subsp. sativus), namely Nantes, Egmont Gold and baby carrots during thermal treatment. Enzyme-substrate reaction kinetics of AAO were described using Michaelis–Menten equation. The estimated K_m and V_max values of AAO ranged from 50.34 to 63.54 μM and 23.70 to 26.82 μmol/min, respectively. Nantes carrots had the lowest AAO activity. On the other hand, Egmont Gold had the highest V_max. AAO activity in all carrot cultivars was stable up to 50 °C and inactivated above 50 °C. Irreversible thermal inactivation of AAO followed first order kinetics (55–70 °C) and the estimated activation energy of the three carrot cultivars situated between 114.33 and 191.45 kJ/mol. Regarding vitamin C stability, thermal treatment at 60–70 °C has resulted in total conversion of l-AA to DHAA due to residual AAO activity; a complete AAO inactivation was found in 80 °C-treated carrots with high vitamin C retention predominantly in l-AA form, up to 90%. On average, the carrots had a total vitamin C content amounting from 368.24 to 379.87 μg/g dry matter and the Nantes carrots had the highest vitamin C content. The effectiveness of rapid inactivation of endogenous AAO via heating (>80 °C, 10 min) prior to matrix disruption gave protection to l-AA towards enzymatic oxidation, thus resulted in a higher vitamin C content and stability in carrots.     

In vitro and in vivo antioxidant properties of ferulic acid: A comparative study with other natural oxidation inhibitors

Ferulic acid exhibits a wide range of therapeutic effects that are attributed to its potent antioxidant capacity. However, in vitro antioxidant properties of ferulic acid have not been elucidated in detail. Evidence that polyphenols, including ferulic acid, act as antioxidants in vivo is also limited. In order to elucidate in more detail the scientific background of antioxidant activities of ferulic acid, we carried out in vitro and in vivo experiments. We focused on superoxide anion scavenging activity, xanthine oxidase inhibition activity, and chain-breaking activity. The combined antioxidant activity from radical scavenging and xanthine oxidase inhibition of ferulic acid was much weaker than that of (−)-epigallocatechin gallate (EGCG) and ascorbic acid. On the other hand, EGCG, ascorbic acid and ferulic acid exhibited chain-breaking activity and prevented ischaemia-reperfusion-associated intestinal injury. Chain-breaking activity may play a contributory role in the protective effect of ferulic acid on oxidative injury in humans and in in vivo studies.     

Steam blanching effect on polyphenoloxidase,peroxidase and colour of mango (Mangifera indica L.) slices

The heat stability of peroxidase (POD) and polyphenoloxidase (PPO) was investigated in mango (Mangifera indica L.) slices, and the relative colour was studied after different steam blanching times. There was complete inactivation after 5 min for POD and 7 min for PPO. Steam blanching of 3 min gave residual activity of 2.85% and 8.33% for PPO and POD, respectively, and when compared with samples blanched for 5 min had no effect on colour over 20 days of storage. Correlation was found between activities of PPO, POD and colour change over 20 days. After 7 min steam blanching the browning index was stable but less than at 3 and 5 min because non-enzymic browning had occurred. This research suggests that yellowness (b) and lightness (L) values contribute positively to the browning index (BI), compared to redness (a).     

Influence of germination techniques on sprout yield,biosynthesis of ascorbic acid and cooking ability,in chickpea (Cicer arietinum L.)

Effects of germination time and illuminations on sprout yield, biosynthesis of ascorbic acid, cooking ability and moisture accumulation in chickpeas were significant (p ? 0.01). Green light had the highest promoting effect on the ascorbic acid level (40.59 mg/100 g) as compared to other illuminations but significantly reduced the sprout yield (188.6 g) as compared to dark, fluorescence and γ-rays illuminations with significantly high sprout yield (196 g) and imbibing moisture (51%). Cooking time was reduced by 43% due to γ-rays in un-soaked seed. Cooking time increased in all treated chickpea samples after 24 h germination and thereafter decreased significantly. Red light significantly increased the cooking time (68.44 min) followed by fluorescent (64.5 min), yellow (61.8 min) and green light (60.9 min). The results indicated that germination of chickpea under green light was an effective process in enhancing ascorbic acid content while dark, fluorescence and γ-rays were effective in promoting sprout growth and to some extent biosynthesis of ascorbic acid.     

Antioxidants in spring leaves of Oxalis acetosella L.

Oxalis acetosella L. is a common, edible wild plant native to the northern hemisphere. The contents of selected antioxidants, and the antioxidant capacity of young and old spring leaves of O. acetosella, were evaluated. The present study reports foliar contents of ascorbic acid, tocopherols, carotenoids, chlorophyll, flavonoids, phenolic acids and total phenolics, and compares the nutritional value of O. acetosella with other cultivated and wild plants. The composition of foliar antioxidants was found to depend on leaf age. On the other hand, the antioxidant capacity of old leaves were in the same range as young leaves. A comparison between O. acetosela with lettuce analysed in our study, and with numerous cultivated and wild edible plants from other studies, showed that O. acetosella is very rich in β-carotene, ascorbic acid, tocopherols and xanthophylls, and that it is one of the best sources of flavonoids (flavonol glycosides and flavan-3-ols), especially rutin. Therefore, O. acetosella is a potentially important dietary source of antioxidants.     

Total phenolics,ascorbic acid,and antioxidant capacity of noni (Morinda citrifolia L.) juice and powder as affected by illumination during storage

Total phenolics, ascorbic acid, and antioxidant capacity of noni (Morinda citrifolia L.) juice and powder were determined during storage at 24 °C. After 2 weeks of storage, illuminated noni juice lost 32% of total phenolics, 89% of ascorbic acid, and 46–65% of antioxidant capacity—about 8%, 22%, and 9–15% more than unilluminated juice. Both illuminated and unilluminated juice lost 97% of ascorbic acid by 4 weeks. The difference in antioxidant characteristics between illuminated and unilluminated juice became insignificant at 12 weeks. After 12 weeks, illuminated noni powder lost 21% of total phenolics, 17% of ascorbic acid, and 23–36% of antioxidant capacity—about 13%, 4%, and 7–19% more than the unilluminated powder. Noni powder in brown bottles retained antioxidant characteristics significantly greater than that in clear bottles. Protection from light effectively reduced degradation of antioxidant characteristics of noni juice for only 2 weeks but those of noni powder for at least 3 months.     

Temperature and pressure stability of mustard seed (Sinapis alba L.) myrosinase

Myrosinase, an enzyme found in all glucosinolate containing plants, is responsible for the conversion of glucosinolates into products that can be beneficial to our health. In this study, the temperature and pressure stability of partially purified myrosinase from mustard seeds was studied in a model system. Temperature inactivation started at 60 °C and the inactivation kinetics were studied in detail between 65 and 75 °C. Inactivation could be described by the consecutive step or the biphasic model. Mustard seed myrosinase was quite pressure stable, as its activity was retained after pressure treatments up to 600 MPa combined with temperatures up to 60 °C. At low pressures there was an antagonistic effect between pressure and thermal treatment, since myrosinase activity was retained after treatments at 70 °C up to 300 MPa. This pressure stability indicates that pressure treatment may be a valuable alternative for thermal treatment if one wants to retain myrosinase activity.     

Fate of Escherichia coli O157:H7, Salmonella and Listeria innocua on minimally-processed peaches under different storage conditions

Consumption of fresh-cut produce has sharply increased recently causing an increase of foodborne illnesses associated with these products. As generally, acidic fruits are considered ‘safe’ from a microbiological point of view, the aim of this work was to study the growth and survival of Escherichia coli O157:H7, Salmonella and Listeria innocua on minimally-processed peaches. The three foodborne pathogens population increased more than 2 log₁₀ units on fresh-cut peach when stored at 20 and 25 °C after 48 h. At 10 °C only L. innocua grew more than 1 log₁₀ unit and it was the only pathogen able to grow at 5 °C. Differences in growth occurred between different peach varieties tested, with higher population increases in those varieties with higher pH (‘Royal Glory’ 4.73 ± 0.25 and ‘Diana’ 4.12 ± 0.18). The use of common strategies on extending shelf life of fresh-cut produce, as modified atmosphere packaging and the use of the antioxidant substance, ascorbic acid (2% w/v), did not affect pathogens’ growth at any of the temperatures tested (5 and 25 °C). Minimally-processed peaches have shown to be a good substrate for foodborne pathogens’ growth regardless use of modified atmosphere and ascorbic acid. Therefore, maintaining cold chain and avoiding contamination is highly necessary.     

Effect of sonication on colour,ascorbic acid and yeast inactivation in tomato juice

Power ultrasound is recognised as a potential non-thermal technique to inactivate micro-organisms pertinent to fruit juices. In this study tomato juice was sonicated at different amplitude levels (24.4–61.0 μm) at a constant frequency of 20 kHz for treatment times (2–10 min) and pulse durations of 5 s on and 5 s off. Hunter colour values (L∗, a∗ and b∗), pH, °Brix, titratable acidity, ascorbic acid and yeast inactivation were measured. No significant differences (p < 0.05) in pH, °Brix or titratable acidity were observed. Regression modelling was used to investigate the main effects of amplitude level and treatment time. Prediction models were found to be significant (p < 0.05) with low standard errors and high coefficients of determination (R²). Model predictions for critical quality parameters of Hunter colour values (L∗, a∗ and b∗), ascorbic acid and yeast inactivation were closely correlated with the experimental results obtained.     

Development of an integrated approach for the stability testing of flavonoids and ascorbic acid in powders

An integrated approach for determining the stability of flavonoids and ascorbic acid in powders was developed. A rapid analyte solubility procedure was used to decide whether HPLC, GLC, or NMR would best serve to measure the stability of each flavonoid. Both the flavonoid under study and ascorbic acid were measured in one analysis with any of the determinative methods. Seventeen flavonoids of differing types, as well as ascorbic acid, were evaluated to establish the approach. The effects of storage relative humidity (RH) on the stability of several flavonoid powders with and without ascorbic acid were then determined. Quercetin, luteolin, taxifolin, naringenin, and naringin were stable for up to 8 weeks in RHs up to 98%, and ascorbic acid was significantly destroyed in the presence of quercetin, luteolin, and taxifolin at 98% RH. Both grape anthocyanins and ascorbic acid were unstable in the presence of each other at 98% RH. The integrated approach could be used for shelf-life testing of powdered flavonoids under a variety of storage conditions.     

Nutraceutical properties and polyphenolic profile of berry skin and wine of Vitis vinifera L. (cv. Aglianico)

Red grapes are rich in phenolics, flavonoids, anthocyanins and resveratrol, all substances which have been suggested as having nutraceutical and health benefits. The berry skin and wine of grape cultivar Vitis vinifera L. (cv. Aglianico), grown in Basilicata (Southern Italy) were examined to determinate the presence of the above mentioned compounds as well as to establish the inorganic cation profile. HPLC analysis coupled with LC–ESI/MS/MS detected high contents of total flavonols and anthocyanins in berry skin and wine. The wine made with the same grape used for berry skin assays showed a notable presence of quercetin-3-O-glucoside (39.4% of total flavonols), and malvidin and petunidin derivatives (63.9% and 10.8% of total anthocyanins, respectively). The strong antioxidant ROS-scavenging activity, determined by both DPPH and FRAP assays, and the high resveratrol content confer high sensory characteristics resulted to be associated with positive nutraceutical properties of these grapes and wine. The level of cis-resveratrol was lower than trans-resveratrol in both berry skin and wine reaching 44.1 mg/kg and 0.3 mg/l, respectively. The cation profile presents low levels of Ca, Cu, K, Fe, Zn and Cd compared to numerous, important red wines, such as Monastrell and Tempranillo.