Monomeric phenols of cashew apple (Anacardium occidentale L.)

Monomeric phenols were extracted by acetone/water (60:40) from the skin and flesh of four cashew apple genotypes from Brazil and Bénin (West Africa), purified by absorption chromatography and subjected to HPLC-DAD/ESI-MS analysis. Skins were found much richer than fleshes in simple phenolics. Flavonol glycosides were dominant with myricetin and quercetin hexosides (2 of each), pentosides (3 of each), and rhamnosides as major compounds. Anthocyanidin glycosides were detected in skins from the two scarlet and orange pigmented genotypes as peonidin, petunidin and cyanidin 3-O-hexosides, and were absent from fleshes.     

Cashew apple (Anacardium occidentale L.) extract from by-product of juice processing: A focus on carotenoids

Cashew apple fibrous residue is a by-product of the cashew juice industry. After pressing using a helical type continuous press followed by crossflow microfiltration, an aqueous extract was obtained from these cashew apple fibres. It was characterised by an intense yellow colour due to carotenoid pigments. Carotenoids were identified and quantified in the cashew apple before extraction, in its aqueous extract and in the concentrate obtained by microfiltration. Cashew apple aqueous extract and its concentrate presented a carotenoid profile with 11 carotenoids, most of them were tentatively identified by HPLC-DAD–MS and are xanthophylls present under an esterified form. Auroxanthin and β-cryptoxanthin represented around 50% of total carotenoids. Concentration of the extract by microfiltration led to epoxy-furanoxy rearrangement of violaxanthin and antheraxanthin. The process allowed an increase of 10 times total carotenoid content compared with initial cashew apple. Total carotenoid content of the final concentrated extract reached 54 mg/kg.     

Characterisation of highly polymerised prodelphinidins from skin and flesh of four cashew apple (Anacardium occidentale L.) genotypes

Tannins were extracted by acetone/water 60:40 from skin and flesh of four cashew apple genotypes from Brazil and Bénin (West Africa), and separated from monomeric phenols. Tannins were submitted to acid-catalysed degradation in the presence of phloroglucinol and the products were analysed by HPLC–DAD/ESI–MS. Both skin and flesh tannins contained high percentages of (-)-epigallocatechin and (-)-epigallocatechin-O-gallate, followed by minor quantities of (-)-epicatechin and (-)-epicatechin-3-O-gallate; 100% of the compounds were the 2,3-cis configuration. Skin tannins were half as galloylated (∼20%) than flesh tannins (∼40%). Their weight-average molecular weight (M_w) was high.     

Functional properties of protein concentrates and isolates produced from cashew (Anacardium occidentale L.) nut

Protein isolates and concentrates were obtained from defatted cashew nut powder by two methods: alkaline extraction-isoelectric precipitation (IP) and alkaline extraction-methanol precipitation (MP). The functional properties of cashew nut protein isolates, concentrates and powder were significantly different (p < 0.05). Cashew nut protein isolate (CNPI) had higher water and oil absorption capacities (2.20 ml/g and 4.42 ml/g, respectively), emulsifying stability index (447%), foam capacity and stability (45% and 55%, respectively), and least gelation capacity (13.5%) than cashew nut protein concentrate (CNPC), which was also higher than that of defatted cashew nut powder (DCNP). However, emulsifying activity index (12.45%) and bulk density (0.31) of CNPI were lower than that of CNPC, which were also lower than that of DCNP. The water solubility of CNPI (95%) and CNPC (95%) was not significantly different (p > 0.05) among the samples, but was significantly different (p < 0.05) from that of DCNP (75%). The CNPI, CNPC and DCNP showed decreasing solubility with decreasing pH, with the minimum solubility being observed at a pH range of 4.0–4.5, confirming the isoelectric point of cashew proteins. However, higher water solubility, emulsifying activity, and foaming property were observed at an alkaline pH than at an acidic pH in all samples.     

Prebiotic effect of fermented cashew apple (Anacardium occidentale L) juice

Leuconostc mesenteroides B-512F and L. mesenteroides B-742 were cultivated in clarified cashew apple juice to produce prebiotic oligosaccharides. Yeast extract (20 g/L); K₂HPO₄ (g/L) and sucrose (50 g/L) were added to the juice to promote the microbial growth and dextransucrase production. Initial pH was adjusted to 6.5 with H₃PO₄. Fermentations were carried out at 30 °C and 150 rpm for 24 h. The prebiotic effect of the fermented cashew apple juice, containing oligosaccharides, was evaluated through the Lactobacillus johnsonii B-2178 growth. L. johnsonii was incubated for 48 h using fermented cashew apple juice as substrate. Lactobacillus growth was compared to the microbial growth in non-fermented juice and in MRS broth. L. johnsonii growth in the fermented cashew apple juice was threefolds the observed growth in the non-fermented juice.     

Determination of the flavonoid components of cashew apple (Anacardium occidentale) by LC-DAD-ESI/MS

Liquid chromatography, with diode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI/MS), was used to identify and quantify flavonoids in cashew apple. One anthocyanin and thirteen glycosylated flavonols were detected in a methanol–water extract. Among them, the 3-O-galactoside, 3-O-glucoside, 3-O-rhamnoside, 3-O-xylopyranoside, 3-O-arabinopyranoside and 3-O-arabinofuranoside of quercetin and myricetin, as well as kaempferol 3-O-glucoside were identified by direct comparison with standards or positively identified flavonoids in cranberry. The anthocyanin was the 3-O-hexoside of methyl-cyanidin. Trace amounts of delphinidin and rhamnetin were detected in the hydrolyzed extract, suggesting their glycosides were present, but undetectable, in the original extract. The concentrations of the 14 flavonoids in the tested sample were determined. This is the first report of these flavonoids in cashew apple.     

The efficacy of cashew nut (Anacardium occidentale L.) skin extract as a free radical scavenger

The free radical scavenging activity of ethanolic extracts of cashew nut (Anacardium occidentale, L.) skin powder (CSP) was evaluated by employing various in vitro antioxidant assay systems. The yield of the extract as well as the total phenolic content was also determined. The yield of ethanolic extract of the skin powder was quite high (0.45 g/g powder) with a total phenolic content of 243 mg/g extract. The cashew nut skin extract (CSE) demonstrated promising antioxidant activity with EC₅₀ of 1.30 ± 0.02 μg/ml in 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assay, 10.69 ± 1.13 μg/ml in superoxide scavenging assay, 17.70 ± 0.05 μg/ml in deoxyribose oxidation assay, 24.66 ± 0.32 μg/ml in lipid peroxidation (LPO) assay and 6.00 mg/ml in iron chelation assay. To identify the compounds in the CSE responsible for the antioxidant activity, thin layer chromatography (TLC) was performed with the extract. The spot showing protection towards β-carotene bleaching was extracted and analyzed by high performance liquid chromatography (HPLC); epicatechin was found to be the major polyphenol present. The results of the present study suggest that cashew nut skin, a byproduct of cashew processing industry, can be used as an economical source of natural antioxidants.     

Catechin and epicatechin in testa and their association with bioactive compounds in kernels of cashew nut (Anacardium occidentale L.)

Catechins in testa and bioactive compounds in testa-free and testa-containing kernels of cashew nuts were analysed. The cashew nut testa contained (+)-catechin and (−)-epicatechin with concentrations of 5.70 and 4.46 g per kg DM, respectively. Testa-containing kernels revealed significantly higher levels of β-carotene (218 vs. 89.6 μg/kg DM), lutein (525 vs. 292 μg/kg DM), and α-tocopherol (10.1 vs. 2.4 mg/kg DM), similar amounts of zeaxanthin (7.0 vs. 7.1 μg/kg DM), γ-tocopherol (10.6 vs. 10.1 mg/kg DM), stearic acid (41 vs. 43 g/kg DM), oleic acid (214 vs. 219 g/kg DM) and linoleic acid (69 vs. 62 g/kg DM), but a lower concentration of thiamine (3.0 vs. 10.7 mg/kg DM) in comparison to testa-free samples. The testa-containing kernels provide high amounts of catechins and higher concentrations of β-carotene, lutein and α-tocopherol than do testa-free cashew nut kernels. This could have potential health benefits for consumers.     

Purification and kinetic characterization of polyphenol oxidase from Barbados cherry (Malpighia glabra L.)

Polyphenol oxidase (PPO) of Barbados cherry was extracted and purified through ammonium sulfate precipitation, gel filtration, and affinity chromatography. The purification factor for PPO was 60% with 8.3% yield. The enzyme was characterized for thermal stability, pH and kinetic parameters. The molecular mass of PPO was approximately the sum of 52 and 38 kDa estimated by SDS–PAGE. The purity was checked by native PAGE, showing a single prominent band. The optimum pH was 7.2. The enzyme had a temperature optimum at 40 °C and was relatively stable at 60 °C, with 55% loss of activity. Sodium diethyl dithiocarbamate (SDDC), l-cysteine and ascorbate significantly inhibited PPO activity. 4-Methyl catechol and catechol were found to be efficient diphenolic substrates for cherry PPO, considering the _Vmax/_Km$V_{\text{max}}/K_{\mathrm{m}}$ ratio. The data obtained in this study may help to understand cherry fruit browning.     

Change of polyphenol oxidase activity, color, and browning degree during storage of cloudy apple juice treated by supercritical carbon dioxide

The effects of supercritical carbon dioxide (SCCO₂) treatment of 8, 15, 22, and 30 MPa for 60 min at 55 °C on polyphenol oxidase (PPO) activity, color, and browning degree in cloudy apple juice during storage at 4 °C for 4-weeks were investigated. The SCCO₂ treatment had significant effects on inactivation of PPO and the least residual activity of PPO was 38.50% at 30 MPa. The restoration of PPO residual activity after SCCO₂ treatment was also observed, which was dependent on the pressure level. A greater reduction of lightness L and a minor increase of redness a of cloudy apple juices after SCCO₂ treatment occurred. Moreover, the total color difference (ΔE), which was significantly less than that of untreated sample, was decreased by enhancing the pressure level. The changes of lightness L and browning degree A during storage were well fitted to a first-order kinetic model. The rate constants of k _L and k _A of samples subjected to SCCO₂ treatment reduced from 4.75×10⁻² to 0.42×10⁻² and 37.19×10⁻² to 8.02×10⁻², respectively, when pressure increased from 0 MPa (untreated sample) to 30 MPa.     

The purification and characterisation of polyphenol oxidase from green bean (Phaseolus vulgaris L.)

Four isoforms of polyphenol oxidases (PPOs) were characterised in purified extracts of coats (PPOIa and PPOIb) and pods (PPOIIa and PPOIIb) of green bean (Phaseolus vulgaris L.). The molecular weights of four isoforms have been estimated to be from 57.5 to 39.0 kDa by SDS–PAGE. The PPOII (mixture of PPOIIa and PPOIIb) was used to characterise the PPO of green bean pods. All isoforms activities were stable between pH 6.8 and pH 7.2. PPOIa and PPOII have similar thermal inactivation profiles, and PPOIb has higher thermal stability than that of PPOIa and PPOII. PPOs showed the highest affinity to pyrogallol in all selected substrates. Although activities of PPOs were markedly inhibited by l-ascorbic acid, the activity of PPOI (mixture of PPOIa and PPOIb) was significantly activated by MnSO₄ and CaCl₂.     

Effect of whey protein concentrate on phenolic profile and browning of fresh-cut lettuce (Lactuca Sativa)

The in vitro kinetics of lettuce PPO with respect to dissolved oxygen using catechin, chlorogenic acid, caffeic acid and gallic acid has been examined. In-vitro lettuce polyphenol oxidase (PPO) activity was determined by measuring the consumption of oxygen during the oxidation reaction. The effect of whey protein concentrate (WPC) was tested on the inhibition of lettuce PPO comparing with ascorbic acid (AA) and cysteine. A competitive model that considered inhibitors was the most appropriate model to explain reaction kinetics. Browning of lettuce was also monitored during storage for 24 h. Addition of WPC prevented loss of lightness in lettuce. Loss of identified phenolic compounds in lettuce was measured during the enzymatic browning process by high-performance liquid chromatography. Degradation of identified phenolic compounds followed first order kinetics during storage. Combination of WPC with cysteine was proposed for the protection of phenolics compounds against PPO-catalysed oxidation.     

Purification and Characterization of Polyphenol Oxidase from Hemşin Apple (Malus communis L.)

The polyphenol oxidase (PPO) enzyme was purified and characterized from Hem?in Apple (Malus communis L.), which was organically grown in Hem?in, in the Rize province of Turkey. Enzyme (PPO) activation was determined with catechol substrate. Apples were homogenized with homogenate buffer (pH 8.5). This process was followed by precipitation with (20–80%) saturated solid (NH₄)₂SO₄ and dialysis. Finally, purification with DE52-Cellulose ion-exchange and Sephadex G-25 columns was performed. Experiments were performed at an optimum pH (5.5) and optimum temperature (30–40°C). The kinetic and thermal parameters Km (3.40 mM), Vmax (333.3 EU/mL.min), Ea (3.57 kcal), ?H (2.968 kcal/mol), Q₁₀ (1.33), k_cat (24.57 min^?1) and V₀ (7.2x10³ mM^?1.min^?1) were assessed. Additionally, the effects of Mg²⁺, Pb ²⁺, Fe²⁺, Fe³⁺, Cd²⁺, Cu²⁺, Zn²⁺, Co²⁺, Al³⁺, Mn²⁺ and Na⁺ on enzyme activity was recorded, and the IC₅₀ values, K_? values and inhibition types were determined.     

Anthocyanins and polyphenol oxidase from dried arils of pomegranate (Punica granatum L.)

Anthocyanins are natural pigments responsible for red, purple, and blue colouration in plants. Human consumption of anthocyanins is increasing because of the rising awareness and interest in their potential health benefits. Pomegranate is one of the major sources of polyphenolic phytochemicals, such as anthocyanins. Dried pomegranate raisins (anardana) are consumed in large quantities in Asian countries, and contain substantial amounts of anthocyanins. Five anthocyanins were found to be present in pomegranate raisins. Drying adversely affected the amount of anthocyanins, with polyphenol oxidase (PPO) playing a possible role in oxidative degradation of anthocyanins. Anthocyanins are heat-stable compounds, and inactivation of PPO by processing at high temperature for short periods may prevent PPO-catalysed anthocyanin oxidation in pomegranate arils. Pomegranate PPO kinetics were partially characterised by investigating the effect of substrate (catechol) concentration; optimum pH for PPO activity was found to be 6.0.     

Inhibitory effect of onion extract on polyphenol oxidase and enzymatic browning of taro (Colocasia antiquorum var. esculenta)

The inhibitory effect of onion extract on polyphenol oxidase and enzymatic browning of taro was investigated. The polyphenol oxidase from taro was strongly inhibited by various reducing agents, such as l-ascorbic acid, l-cysteine, dithiothreitol, glutathione and sodium pyrosulfite. The enzyme was also inhibited by addition of onion extract. Regardless of substrates used, the addition of heated onion extract at 100 °C for 10 min, gave a stronger inhibitory effect on taro polyphenol oxidase activity than did fresh unheated extrtact. The inhibitory effect of onion extract was dependent on heating temperature and time. The addition of glucose, glycine, or both to the onion extract, during heating, stimulated the inhibitory effect of the onion extract, suggesting that non-enzymatic browning products, produced during heating, might be responsible for the stronger inhibitory action of the heated onion extract.     

Sugars,organic acids,phenolic composition and antioxidant activity of sweet cherry (Prunus avium L.)

Sugars, organic acids, phenolics and anthocyanins in fruits of 13 sweet cherry cultivars: Badascony, Burlat, Early Van Compact, Fercer, Fernier, Ferprime, Lala Star, Lapins, Noire de Meched, Sylvia, Vesseaux, Vigred (red-coloured) and Ferrador (bi-coloured) were quantified by HPLC. Sweet cherry cultivars of different pomological characteristics and different time of ripening were evaluated sensorily. Cultivars were evaluated for their total phenolic content and antioxidant activity. The sum of sugars (glucose, fructose, sucrose and sorbitol) ranged from 125 to 265 g/kg fresh weight (FW) and the sum of organic acids (malic, citric, shikimic and fumaric) ranged from 3.67 to 8.66 g/kg FW. Total phenolic content ranged from 44.3 to 87.9 mg gallic acid equivalents/100 g FW and antioxidant activity ranged from 8.0 to 17.2 mg ascorbic acid equivalent antioxidant capacity mg/100 g FW. The correlation of antioxidant activity with total phenolics content and content of anthocyanins was cultivar dependent.     

Polyphenol oxidase from bayberry (Myrica rubra Sieb. et Zucc.) and its role in anthocyanin degradation

Polyphenol oxidase (PPO) was extracted from bayberry (Myrica rubra Sieb. et Zucc. cv. Biqi), and its partial biochemical characteristics were studied. Stable and highly active PPO extracts were obtained using insoluble polyvinylpolypyrrolidone (PVPP) in sodium phosphate, pH 7.0, buffer. The highest PPO activity was observed in the ripe fruits. Optimum pH and temperature for bayberry PPO activity were pH 6.0 and T = 30 °C with 0.1 M catechol as substrate. PPO showed activity using the substrates of catechol, gallic acid and protocatechuic acid, but no activity with the substrates (+)-catechin, p-coumaric acid or caffeic acid. K_m and V_max values were 313 mM and 3.26ΔOD/min/g FW, respectively, with catechol as the substrate. Bayberry PPO did not act directly on cyanidin 3-glucoside but the rate of anthocyanin degradation was stimulated by the addition of gallic acid.     

Enzymatic browning and after-cooking darkening of Jerusalem artichoke tubers (Helianthus tuberosus L.)

Jerusalem artichoke tubers (Helianthus tuberosus L.) undergo enzymatic browning when peeled or cut, and turn grey after boiling, due to after-cooking darkening reactions between iron and phenolic acids. In an attempt to reveal the components responsible for these discolouration reactions, sensory evaluation and instrumental colour measurements were related to contents of total phenolics, phenolic acids, organic acids and iron in three varieties of raw and boiled Jerusalem artichoke tubers harvested in the autumn and the spring. No differences were found between varieties in sensory evaluated enzymatic browning, but Rema and Draga had higher scores than Mari in after-cooking darkening. Jerusalem artichoke tubers had higher contents of total phenolics, phenolic acids and citric acid in the autumn and low contents in the spring, while it was the opposite for malic acid. None of the chemical parameters investigated could explain the discolouration of the Jerusalem artichoke tubers.     

Improvement of frozen banana (Musa cavendishii, cv. Enana) colour by blanching: relationship between browning, phenols and polyphenol oxidase and peroxidase activities

 Browning in banana (Musa cavendishii, cv. Enana) processed products is a result of phenol oxidation catalysed by polyphenol oxidase (PPO) and peroxidase (POD) or of other non-enzymatic reactions (Maillard and Strecker mechanisms). Microwave and steam blanching significantly reduced PPO and POD activities and phenol levels in banana flesh, steam blanching being the most effective method for enzyme inactivation. Freezing/thawing processes produced a significant increase in phenol levels in all samples, due to cellular breakdown. After microwave heating browning processes occurred while steam-treated samples did not exhibit a significant colour change. Extractable PPO and POD activities in all banana samples increased as a consequence of freezing/thawing: steam-blanched slices exhibited lower residual activities. High correlations occurred between phenols and browning (r=0.86) in control samples. Blanched samples (microwave or steam) only exhibited correlations between PPO (r=0.80) and POD (r=0.80) activities and browning. Received: 22 February 1996     

Improvement of frozen banana (Musa cavendishii, cv. Enana) colour by blanching: relationship between browning, phenols and polyphenol oxidase and peroxidase activities

 Browning in banana (Musa cavendishii, cv. Enana) processed products is a result of phenol oxidation catalysed by polyphenol oxidase (PPO) and peroxidase (POD) or of other non-enzymatic reactions (Maillard and Strecker mechanisms). Microwave and steam blanching significantly reduced PPO and POD activities and phenol levels in banana flesh, steam blanching being the most effective method for enzyme inactivation. Freezing/thawing processes produced a significant increase in phenol levels in all samples, due to cellular breakdown. After microwave heating browning processes occurred while steam-treated samples did not exhibit a significant colour change. Extractable PPO and POD activities in all banana samples increased as a consequence of freezing/thawing: steam-blanched slices exhibited lower residual activities. High correlations occurred between phenols and browning (r=0.86) in control samples. Blanched samples (microwave or steam) only exhibited correlations between PPO (r=0.80) and POD (r=0.80) activities and browning. Received: 22 February 1996