Study of the inhibitory activity of phenolic compounds found in olive products and their degradation by Lactobacillus plantarum strains

Lactobacillus plantarum is the main species responsible for the spontaneous fermentation of Spanish-style green olives. Olives and virgin oil provide a rich source of phenolic compounds. This study was designed to evaluate inhibitory growth activities of nine olive phenolic compounds against four L. plantarum strains isolated from different sources, and to explore the L. plantarum metabolic activities against these phenolic compounds. None of the nine compounds assayed (oleuropein, hydroxytyrosol, tyrosol, as well as vanillic, p-hydroxybenzoic, sinapic, syringic, protocatechuic and cinnamic acids) inhibited L. plantarum growth at the concentration found in olive products. Oleuropein and tyrosol concentrations higher than 100 mM were needed to inhibit L. plantarum growth. On the other hand, sinapic and syringic acid showed the highest inhibitory activity since concentrations ranging from 12.5 to 50 mM inhibited L. plantarum growth in all the strains analyzed. Among the nine compounds assayed, only oleuropein and protocatechuic acid were metabolized by L. plantarum strains grown in the presence of these compounds. Oleuropein was metabolized mainly to hydroxytyrosol, while protocatechuic acid was decarboxylated to catechol. Metabolism of oleuropein was carried out by inducible enzymes since a cell-free extract from a culture grown in the absence of oleuropein was unable to metabolize it. Independent of their isolation source, the four L. plantarum strains analysed showed similar behaviour in relation to the inhibitory activity of phenolic compounds, as well as their ability to metabolize these compounds.     

Phenolic compounds in virgin olive oils: influence of the degree of olive ripeness on organoleptic characteristics and shelf-life

 An experimental investigation was performed on virgin olive oils from Coratina and Ogliarola salentina cultivars extracted from green, partially blackened and totally blackened olives in order to evaluate changes in the organoleptic properties and in the shelf-life of the oil. The aim of the investigation was to identify the optimal stage of olive maturation for each cultivar to produce higher quality oils. Routine analyses in accordance with EC regulation no. 2568/91, the determination of phenols by HPLC and induction times were carried out. On the whole, the simple phenolic compounds increased as darker olives were used whereas the hydrolysable phenolic compounds decreased. In both the cultivars, total phenols and induction times were significantly higher in the oils obtained from green olives than in oils from totally blackened olives. Hence, it seems advisable to delay olive harvesting for cultivars usually yielding bitter to pungent oils (e.g. Coratina), but to anticipate harvesting for olives producing sweet tasting oils (e.g. O. Salentina). This would produce less bitter Coratina oils, with organoleptic features which may better fulfil consumers' expectations, and O. Salentina oils more resistant to oxidation. Received: 5 June 2000     

Determination of phenolic compounds of ‘Sikitita’ olive leaves by HPLC-DAD-TOF-MS. Comparison with its parents ‘Arbequina’ and ‘Picual’ olive leaves

‘Sikitita’ is a new olive cultivar developed in Spain by crossing the cultivars ‘Picual’ and ‘Arbequina’. The phenolic fraction of olive leaves (Olea europaea L.), is purported to have good anti-oxidative properties which help to prevent several health problems. To our knowledge, no studies are available on the phenolic fraction of ‘Sikitita’ olive leaves or any other new cultivar developed from breeding. Thus, the identification and quantification of the phenolic fraction of ‘Sikitita’ olive leaves by HPLC-DAD-MS were studied and compared with those of its parent cultivars. The three cultivars were grown under the same agronomic and environmental conditions in the same orchard. The quantification was performed using HPLC-DAD, whereas qualitative data were acquired using HPLC-MS. It was thus possible to identify 30 different compounds, two of which have been tentatively characterized for the first time in olive leaves of Spanish cultivars. Significant differences between cultivars were observed for almost all the compounds. Results for ‘Sikitita’ olive leaves presented a higher degree of similarity with respect to ‘Picual’ than to ‘Arbequina’. Further work will monitor the time course of phenolic compounds over the growth period.     

Influence of air temperature on drying kinetics and antioxidant potential of olive pomace

This work aims to evaluate the influence of olive pomace drying (a solid by-product of the olive oil industry) on both antioxidant potential and drying kinetics. The two main fractions of olive pomace (pits, PI and pulps + peels, P + P) were characterized by image analysis and density measurement. The drying process was analyzed in experiments carried out at different temperatures (from 50 to 150 °C) and mathematically described from the diffusion and Weibull models. The antioxidant potential of the extracts (ethanol–water 80:20 v/v, 22 ± 1 °C, 170 rpm for 24 h) obtained from the dry product was analyzed by measuring the total phenolic content and antioxidant capacity and the main polyphenols were quantified by HPLC–DAD/MS–MS.     

Comparison of methods extracting phenolic compounds from lyophilised and fresh olive pulp

This report’s general objective is to propose a method for extracting the polyphenols in lyophilised olive pulp. We studied the qualitative profiles of the corresponding phenolic extracts, using samples of Picual, Hojiblanca and Arbequina olive varieties, in order to compare the proposed method with traditional fresh pulp methods. The proposed method, based on the extraction of lyophilised olive pulp with a mixture of ethanol-water (80:20 v/v), resulted in less hydrolysis of oleuropein and ligstroside, and the extract was obtained in a 33% shorter time using 30% less solvent.     

Impact of oregano and virgin olive oil phenolic compounds on acrylamide content in a model system and fresh potatoes

In this study the effect of olive oil and oregano phenolic compounds on antioxidant activity (AOA) and acrylamide formation in a model system and fresh potatoes was evaluated. The addition of oregano phenolic extract in the model system resulted in an increase of AOA and in a reduction of acrylamide content, up to 49%, after heating. On the contrary, addition of olive oil extract resulted in AOA reduction and a concentration-dependent effect on acrylamide formation was observed with a reduction up to 15% (p < 0.05) followed by an increase up to 48%. The same trend, in acrylamide formation, was observed in fresh potatoes but to a lower extent. Finally, addition of tyrosol, oleuropein and p-hydroxyphenylacetic acid in the model system resulted in acrylamide reduction up to 50%. The discrepancy in acrylamide formation, caused by phenolic compounds, is ascribed to their structure and related to their terminal functional hydroxyl and aldehydic groups.     

Effect of salinity stress on phenolic compounds and carotenoids in buckwheat (Fagopyrum esculentum M.) sprout

The effect of salinity stress on the nutritional quality of buckwheat sprouts cultivated for 1, 3, 5, and 7d was investigated by analysis of the antioxidant activity and levels of phenolic compounds and carotenoids. Treatment with various concentrations of NaCl (10, 50, 100, and 200mM) resulted in an increase in the amount of phenolic compounds and carotenoids in the sprouts compared with the control (0mM). The phenolic contents of sprouts treated with 10, 50, and 100mM after 7d of cultivation were 57%, 121%, and 153%, respectively, higher than that of the control (0mM NaCl). Moreover, the accumulation of phenolic compounds was primarily caused by an increase in the levels of 4 compounds: isoorientin, orientin, rutin, and vitexin. The carotenoid content of sprouts treated with 50 and 100mM NaCl was twice higher than that of the control. In addition, the antioxidant activity of ethanol extracts of the sprouts was increased by NaCl treatment. Although the growth rate of sprouts decreased with >50mM NaCl, these results suggest that treatment of an appropriate concentration of NaCl improves the nutritional quality of sprouts, including the level of phenolic compounds, carotenoids, and antioxidant activity.     

Effect of phenolic compounds on the properties of porcine plasma protein-based film

Properties of porcine plasma protein-based film incorporated with tannic acid, caffeic acid and ferulic acid at different concentrations (1–3% (w/w) of protein content) were studied. Film-forming solution (FFS) containing 3% protein (w/v) and 70% glycerol (w/w of protein content) was preheated at 70 °C for 30 min and adjusted pH to 10 followed by the addition of phenols and film casting. Tensile strength (TS) of resulting film increased by 123.3, 194.3 and 19.5% and elongation at break (EAB) increased by 71.1, 86.3 and 10.2%, respectively, compared with the control film, when tannic acid, caffeic acid and ferulic acid at a level of 3% was added. The use of all phenolic compounds slightly increased water vapor permeability (WVP) of resulting films (p < 0.05). The increases in a^∗- and b^∗-values of films were observed as the higher concentrations of tannic acid and caffeic acid were used. This was associated with the lowered transparency of resulting films. FFS containing 3% caffeic acid with prior oxygenation, especially with pH 10, yielded the film with increased TS but lowered EAB (p < 0.05). Oxygenation of FFS was associated with the lower L^∗-value and higher a^∗-value of resulting films. Therefore, phenolic compounds could be used as natural cross-linkers which affected the properties of porcine plasma protein-based film differently.     

Solvent effects on phenolic contents and biological activities of the halophyte Limoniastrum monopetalum leaves

Limoniastrum monopetalum is a traditional medicinal species which leaf infusion exhibits antidysenteric properties against infectious diseases. In this study, ten kinds of leaf extracts were used to examine the effect of extraction solvent system with varying polarities on polyphenol contents and DPPH scavenging activity. Then the superoxide scavenging activity and the reducing power of the most promising solvent extracts were evaluated too. Moreover, the efficiency of the best leaf extract has been investigated against pathogenic bacteria and yeast. Eventually leaf extract was hydrolyzed by acid and the phenolics identified by RP-HPLC. Results showed that phenolic contents and antioxidant activities varied considerably as function of solvent polarity. Leaf extract using pure methanol showed the highest polyphenol content (15.85 mg GAE/g DW). Moreover, antiradical capacities against DPPH and superoxide, and reducing power were maxima in acetone/water (8:2) of leaf extract. However, the latter showed a slight antimicrobial activity against human pathogen strains such as Staphylococcus aureus, Micrococcus luteus and Candida holmii. The HPLC analysis revealed several phenolic compounds in L. monopetalum leaf including vanillic and gallic acids as major phenolics. Our findings identified the appropriate solvent for extracting halophyte phenolics which might provide a rich and novel source of natural antioxidants as food additives replacing synthetic ones in food industry.     

Antioxidant phenolic compounds loss during the fermentation of Chétoui olives

Evolution of phenolic compounds was studied during spontaneous and controlled fermentations of “Chétoui” cultivar olives at three degree of ripeness. Both spontaneous and controlled fermentations led to an important loss of total phenolic compounds with a reduction rate of 32–58%. Consequently, the antioxidant activity decreased by 50–72%. After fermentations, hydroxytyrosol and caffeic acid concentrations increased, whilst protocatechuic acid, ferulic acid and oleuropein concentrations decreased. The hydroxytyrosol concentration in black olives increased from 165 mg/100 g dry weight to 312 and 380 mg/100 g dry weight, respectively, after spontaneous and controlled fermentation. The oleuropein concentration in green olives decreased from 266 mg/100 g dry weight to 30.7 and 16.1 mg/100 g dry weight, respectively, after spontaneous and controlled fermentation. During olive fermentation, phenolic loss is essentially due to the diffusion of these compounds into the brine; the main phenolic compound identified and quantified in the different brines was hydroxytyrosol. To preserve antioxidant quality of table olives it is necessary to use a controlled process to minimise phenolic compound loss.     

Determination of free phenolic acids and antioxidant activity of methanolic extracts obtained from fruits and leaves of Chenopodium album

In this study, determination of phenolic acids as well as investigation of antioxidant activity of methanolic extracts from the fruits and leaves of Chenopodium album is described. Extracts were subjected to acidic hydrolysis in order to obtain total free phenolic acids. However, some of phenolic acids were identified and quantified by HPLC-DAD. The results were confirmed by LC-MS equipped with MS-ESI. In addition, Folin–Ciocalteu method was applied to determine the total phenolic contents. The antioxidant activity of C. album extracts was examined by using DPPH and hydroxyl radical-scavenging activity assays. Results revealed that the leaves extract exhibits better performance in antioxidant assays and in the higher total phenolic contents (3066 mg of GAE/100 g) when compared to fruits extract (1385 mg of GAE/100 g). From these results it has been revealed that the methanolic extracts of C. album from fruits and leaves have great potential as a source for natural health products.     

Antioxidant activity and phenolic compounds of fractions from Portulaca oleracea L.

Portulaca oleracea fractions obtained from the crude extract of the plant by reversed-phase separation were investigated for their antioxidant activities and phenolic compounds content. Five fractions were classified based on optical absorption between the wavelength range of 200–400 nm. Fraction 3 displayed higher absorption than the other fractions. The total amount of quantified phenolics in this fraction was found to be quite high compared to that of crude extract. Chlorogenic, caffeic, p-coumaric, ferulic and rosmarinic acids were the free phenolic acids, and quercetin and kaempferol were the free flavonoids detected in Fraction 3. IC₅₀ value of crude extract was found to be 511.8 μg/ml whereas Fraction 3 exhibited an IC₅₀ value of 154.1 μg/ml. TEAC of Fraction 3 was found to be almost four times higher than that of the crude extract. Fraction 3 was also found to show the highest lipid peroxidation inhibiting capacity, determined by TBARS assay.     

Antioxidant activity and phenolic contents of Olea europaea L. leaves sprayed with different copper formulations

Olive trees (Olea europaea L. Cv. Cobrançosa) from the northeast of Portugal were sprayed with three different copper formulations [bordeaux mixture (copper sulphate + calcium hydroxide − 20% Cu), copper hydroxide (40% Cu) and copper oxychloride (50% Cu)] to control olive fungal diseases. The residues of copper in olive leaves, harvested at different times, were evaluated by atomic absorption spectrometry. At all the collection times, treated olive leaves had significantly higher copper contents, compared to the control. The different copper amounts in pesticide formulations lowered the leaves contents in total phenols and hence their antioxidant properties. Olive leaves sprayed with copper oxychloride possessed the highest copper levels and the lowest content in phenols, which influenced its antioxidant activity (higher EC₅₀ values for reducing power, scavenging effect on DPPH radicals and inhibition of erythrocyte hemolysis). Leaves without copper residues proved to be a good natural source of antioxidants, giving values comparable to the reference compounds.     

Antioxidant activity and phenolic compounds of Holotrichia parallela Motschulsky extracts

Insects have been relatively unexplored as potential sources of natural antioxidants. We report the antioxidant activity of extracts of the adult large black chafer beetle Holotrichia parallela Motschulsky, a common crop pest in China. The antioxidant activity of the ethanolic extract (EE) and the water extract (WE) of adult H. parallela were evaluated by four different in vitro assays. EE showed potent metal-chelating activity and inhibition of lipid peroxidation. WE proved to be an excellent antioxidant in the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and metal-chelating activity. Catechin was identified in the ethanolic extract and proteins were the main components in the water extracts. Both compounds could contribute to the antioxidant activity of the species. These results suggest that adult H. parallela might be used as a nutraceutical to alleviate oxidate-induced diseases and as a natural antioxidant additive in the food industry.     

Sacha inchi (Plukenetia volubilis): A seed source of polyunsaturated fatty acids,tocopherols, phytosterols,phenolic compounds and antioxidant capacity

Fatty acids (FA), phytosterols, tocopherols, phenolic compounds, total carotenoids and hydrophilic and lipophilic ORAC antioxidant capacities were evaluated in 16 cultivars of Sacha inchi (SI) seeds with the aim to valorise them and offer more information on the functional properties of SI seeds. A high α linolenic (α-Ln) fatty acid content was found in all cultivars (ω3, 12.8–16.0 g/100 g seed), followed by linoleic (L) fatty acid (ω6, 12.4–14.1 g/100 g seed). The ratio ω6/ω3 was within the 0.83–1.09 range. γ- and δ-tocopherols were the most important tocopherols, whereas the most representative phytosterols were β-sitosterol and stigmasterol. Contents of total phenolics, total carotenoids and hydrophilic and lipophilic antioxidant capacities ranged from 64.6 to 80 mg of gallic acid equivalent/100 g seed; from 0.07 to 0.09 mg of β-carotene equivalent/100 g of seed; from 4.3 to 7.3 and, from 1.0 to 2.8 μmol of Trolox equivalent/g of seed, respectively, among the evaluated SI cultivars. Results showed significant differences (p < 0.05) among the evaluated SI cultivars in the contents of ω3, ω6, antioxidant capacities and other evaluated phytochemicals. SI seeds should be considered as an important dietary source of health promoting phytochemicals.     

Chemical composition, nutritional value, and antioxidant constituents of Kalopanax pictus leaves

This study was carried out to analyze the nutritional composition, mineral content and antioxidant activity of Kalopanax pictus leaves. K. pictus leaves are high in carbohydrates (60.37%) and crude protein content (27.05%) on a dry weight basis with low content of fatty acids (1.45%). Hyperoside (1), 3,5-di-O-caffeoyl quinic acid (2), methyl 3,5-dicaffeoyl quinate (3), and 3-O-feruloylquinic acid (4) were isolated and purified from this plant as well. In all the chemical assays used, compound 3 exerted better 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, reducing power ability, and was a potent inhibitor of hydrogen peroxide (H₂O₂)-induced human embryonic kidney 293 (HEK 293) cell death and lipid peroxidation, whereas compound 4 presented as a powerful radical scavenger against superoxide radicals. Our results suggest that the chemical components of K. pictus leaves possess good antioxidant capacities and this species might have potential applications in the food and medical industries.     

Infusion and decoction of wild German chamomile: Bioactivity and characterization of organic acids and phenolic compounds

Natural products represent a rich source of biologically active compounds and are an example of molecular diversity, with recognised potential in drug discovery. Herein, the methanol extract of Matricaria recutita L. (German chamomile) and its decoction and infusion (the most consumed preparations of this herb) were submitted to an analysis of phytochemicals and bioactivity evaluation. The antioxidant activity was determined by free radicals scavenging activity, reducing power and inhibition of lipid peroxidation; the antitumour potential was tested in human tumour cell lines (breast, lung, colon, cervical and hepatocellular carcinomas), and the hepatotoxicity was evaluated using a porcine liver primary cell culture (non-tumour cells). All the samples revealed antioxidant properties. The decoction exhibited no antitumour activity (GI₅₀ > 400 μg/mL) which could indicate that this bioactivity might be related to compounds (including phenolic compounds) that were not extracted or that were affected by the decoction procedure. Both plant methanol extract and infusion showed inhibitory activity to the growth of HCT-15 (GI₅₀ 250.24 and 298.23 μg/mL, respectively) and HeLa (GI₅₀ 259.36 and 277.67 μg/mL, respectively) cell lines, without hepatotoxicity (GI₅₀ > 400 μg/mL). Infusion and decoction gave higher contents of organic acids (24.42 and 23.35 g/100 g dw). Otherwise, the plant methanol extract contained the highest amounts of both phenolic acids (3.99 g/100 g dw) and flavonoids (2.59 g/100 g dw). The major compound found in all the preparations was luteolin O-acylhexoside. Overall, German chamomile contains important phytochemicals with bioactive properties (mainly antitumour potential selective to colon and cervical carcinoma cell lines) to be explored in the pharmaceutical, food and cosmetics industries.     

电子束辐照降解玉米中黄曲霉毒素B1及对玉米品质的影响

为考察电子束辐照对玉米中黄曲霉毒素B_1(aflatoxin B_1,AFB_1)的降解效果及对玉米品质的影响,试验选取AFB_1污染的玉米为原料,研究0~50kGy剂量辐照下玉米中AFB_1的降解效果,并考察电子束辐照对玉米理化性质,包括脂肪酸值、黏度值和色度的影响。结果表明,随着电子束辐照剂量的升高(5~50kGy),玉米中AFB_1含量显著降低(P0.05);当电子束辐照剂量大于15kGy时,玉米理化性质变化较明显,其中脂肪酸值显著升高(P0.05),黏度值显著下降(P0.05),L*值无显著变化(P0.05),a*,b*值显著下降(P0.05)。     

Ultraviolet irradiation: The generator of Vitamin D2 in edible mushrooms

Fresh Shiitake mushrooms (Lentinula edodes), Oyster mushrooms (Pleurotus ostreatus), Button mushrooms (Agaricus bisporus), and Abalone mushrooms (Pleurotus cystidus) were irradiated with Ultraviolet-A (UV-A; wavelength 315–400 nm), Ultraviolet-B (UV-B; wavelength 290–315 nm), and Ultraviolet-C (UV-C; wavelength 190–290 nm). Irradiation of each side of the mushrooms for 1 h, was found to be the optimum period of irradiation in this conversion. The conversions of ergosterol to vitamin D₂ under UV-A, UV-B, and UV-C were shown to be significantly different (p < 0.01). The highest vitamin D₂ content (184 ± 5.71 μg/g DM) was observed in Oyster mushrooms irradiated with UV-B at 35 °C and around 80% moisture. On the other hand, under the same conditions of irradiation, the lowest vitamin D₂ content (22.9 ± 2.68 μg/g DM) was observed in Button mushrooms.