Enrichment of table olives with polyphenols extracted from olive leaves

The possibility to increase the nutritional value of table olives using polyphenols extracted from olive leaves was studied. Leaves were subjected to extraction using water with proportions of 1%, 5% and 10% of leaves and various temperatures and times (room temperature/24 h, 40 °C/10 min and 70 °C/5 min). The antioxidant activity of extracts was determined using the Rancimat method and their content in oleuropein and hydroxytyrosol was determined by HPLC. The extract with the highest antioxidant activity and polyphenol content (10% ratio of olive leaves extracted at room temperature for 24 h) was used for the treatment of debittered table olives. The treated and untreated olives were subjected to determination of their α-tocopherol, oleuropein and hydroxytyrosol content. A 457% increase was observed for oleuropein and 109% for hydroxytyrosol content after treatment. Sensory evaluation of treated table olives showed an increase in bitterness. However, treated and untreated table olives showed equal overall acceptability and overall preference.     

Phenolic and antioxidant potential of olive oil mill wastes

Olive oil mill waste was subjected to conventional liquid solvent extraction and supercritical fluid extraction using different solvents and carbon dioxide, respectively. The optimum solvent extraction conditions of phenols were 180 min using ethanol, at a solvent to sample ratio 5:1 v/w, and at pH 2. Solvent and SFE extracts were tested for their antioxidant activity by the DPPH radical scavenging method and by determination of peroxide value on virgin olive oil and sunflower oil. The ethanol extract exhibited the highest antiradical activity, and no correlation was found between antiradical activity and phenol content. The SFE extract exerted good antioxidant capacity although its phenolic yield was not quite high. Moreover, the ethanol extract appeared to be a stronger antioxidant than BHT, ascorbyl palmitate and vitamin E by the Rancimat method on sunflower oil. HPLC analysis of the extracts showed that the predominant phenolic compound was hydroxytyrosol. Various phenolic acids and flavonoids were also identified.     

High production of Aspergillus niger β‐glucosidase at pilot‐scale and application for hydroxytyrosol release from olive by‐product

This study reports a very simple and fast method to collect extract with high amounts of hydroxytyrosol, biotransforming Olea europaea L. leaf extract and olive mill wastewater by a β‐glycosidase produced locally by Aspergillus niger. These extracts are considered as natural substances from a vegetal source; possessing a higher concentration of simple phenols, characterised by a stronger antioxidant capacity which could be commercially found as dietetic products and food integrators. The proposed process may prove useful for a further application for recycling O. europaea by‐product. In this study, a β‐glucosidase for A. niger was optimised in submerged culture conditions and produced at pilot‐scale. The optimal culture conditions for extracellular β‐glucosidase production by the submerged fermentation of A. niger were investigated using various parameters, including different substrate, nitrogen source, C/N ratio.     

Fatty acid profile,sugar composition,and antioxidant compounds of table olives as affected by different treatments

The influence of cultivar and processing on olive antioxidants, fatty acids, and sugars profiles were studied for the autochthonous Meski and two introduced table olives (Picholine and Manzanella). Olive fruits were treated with two traditional Tunisian processes. Fatty acid analysis by capillary gas chromatography, total phenols and o-diphenols, and sugars profiles by GC and GC–MS, together with oxidative capacity, were evaluated. Independently of the processing method, the olive fruit showed significant cultivar dependant differences. Concerning the effect of processing, ANOVA tests showed no significant difference in the fatty acid profiles, whereas sugars and phenols underwent a sharp decrease during the fruit storage.     

Stability and antioxidant potential of purified olive mill wastewater extracts

A purified olive extract (POE) rich in phenolic and oleosidic compounds was prepared from olive mill wastewaters by adsorption onto an amphoteric polymer resin with the yield of 2.2% (w/v). In addition to complex and simple phenolic compounds found in the POE, the highly hydrophilic elenolic acid and DEDA (dialdehydic form of decarboxymethyl elenolic acid, a non-phenolic secoiridoids) were also present in high concentration (45%, w/w). Among the phenolic constituents, the major compounds are hydroxytyrosol and its glucoside, DEDA linked to hydroxytyrosol (Hy-DEDA), p-coumaric acid and caffeic acid, verbascoside and some flavonoids. The high content (23%, w/w in gallic acid equivalent) and structural diversity of these phenolic compounds confer superior antioxidant potential of POE with an IC₅₀ less than 8.5 μg/ml determined by DPPH assay. The stability of bioactive components during storage of the extract and changes in antioxidant properties were studied under different conditions. Determination of kinetics of degradation revealed that air/oxygen is the determinant factor which influences the stability of POE under low temperature storage conditions, while exposure to sunlight did not have significant effect on their stability. Whilst an increase in storage temperature decreased the total content of phenolic compounds (20-24% reduction) and non-phenolic secoiridoid aglycons, a high antioxidant capacity was still observed particularly in the nitrogen-protected POE samples (93-95% of the initial value). This was believed to be due to the transformation of hydroxytyrosol-containing complex phenols, especially the secoiridoid ester Hy-DEDA, into free phenolic monomer. This contributed significantly to the maintenance of a high antioxidant potency of the whole extract during storage. The high antioxidant, radical scavenging activity and stability of the extract suggest potential applications in the food and pharmaceutical industries.     

Simple phenolic content in olive oil residues as a function of extraction systems

Olive oil residues were tested for their composition in simple phenolic compounds as a function of the extraction system, i.e. the three- and two-phase centrifugation systems. Phenolic compound extraction with ethyl acetate was efficient and allowed recovery of 28.8 and 42.2% of total phenols present in dry olive oil residues originating from three-phase and two-phase systems, respectively. The qualitative and quantitative HPLC analyses of the extracts showed that hydroxytyrosol and p-tyrosol were the most abundant phenolic compounds. p-coumaric, caffeic, ferulic and vanillic acids were also present. The phenolic extract from the two-phase system had the highest concentration in hydroxytyrosol (1.16% (w/w) dry residue) and the strongest antioxidant activity. Olive oil residues were confirmed as a cheap source of large amounts of natural phenolic antioxidants.     

Effect of storage on refined and husk olive oils composition: Stabilization by addition of natural antioxidants from Chemlali olive leaves

Two samples of refined olive and husk oils have been analysed in order to evaluate the influence of storage time on their quality. The following parameters were determined: peroxide values, absorption coefficients K270 and K232, Rancimat induction time, sterols and fatty acid contents. Six months storage at 50 °C in the dark revealed a loss in oil stability. This finding was reflected by the greater increase in peroxide value and a decrease of Rancimat induction time and sterol content. The enrichment of refined olive and husk oils with olive leaves and its hydrolysate extract resulted in an appreciable resistance to oxidative deterioration due to its phenolic antioxidants content. Oleuropein and hydroxytyrosol were the major compounds in Chemlali olive leaves extract and hydrolysate solution, respectively. The antiradical activity of leaves extract as well as its hydrolysate solution was evaluated and compared to that of the BHT. The antioxidant activity of the enriched refined olive and husk oils with leaves and hydrolysate extracts at 400 ppm showed that the latter had the highest protective effect against oil oxidation. Oils with added hydrolysate extract had the lower peroxide value and the higher stability measured with a Rancimat method. After six months of storage the induction time increased from 23.3 to 83.5 h for refined olive oil and from 16.6 to 49 h for husk oil. Furthermore, during oil storage, there was no significant variation in fatty acid composition. However, the total sterol concentration of the oils treated with hydrolysate extract increased. The results suggested that hydrolysate and leaves extracts are excellent antioxidants and can serve as substitutes for synthetic antioxidants.     

Evaluating two kinds of centrifuged virgin oils arising from continuous olive processing

Oils extracted from olive pastes by the direct centrifugation mode were compared with the homologous oils produced by the indirect centrifugation (after percolation) mode. The former were characterised by: (i) higher contents of total phenols, o‐diphenols, hydroxytyrosol, hydroxytyrosol‐aglycons, total volatiles, trans‐2‐hexanal and other pleasant volatiles, total tocopherols, total sterols and waxes; (ii) lower contents of triterpene dialcohols, aliphatic and triterpene alcohols, chlorophylls and pheophytins; (iii) lower values of integral colour index; (iv) higher values of turbidity, campesterol/stigmasterol ratio, 1,2‐diglycerides/1,3‐diglycerides ratio, oxidative stability and overall quality indices; and (v) higher sensory score. Stigmastadienes and trans‐isomer C₁₈ fatty acids were always not detected. The average oil outputs of the two centrifugation extraction procedures were comparable, as confirmed by similar overall oil amounts found in the by‐products. © 2000 Society of Chemical Industry     

Phenylalanine ammonia‐lyase and phenolic compounds in leaves and fruits of Olea europaea L. cv. Picual during ripening

BACKGROUND: The kinetic and molecular properties of phenylalanine ammonia‐lyase (PAL) in leaves and fruit of the olive tree (Picual variety) have been studied during the seasonal process of fruit maturation. The concentrations of total phenolic compounds, oleuropein, hydroxytyrosol and tyrosol, have also been determined. This study has been made in rainfed 30‐year‐old olive trees in Jaén, Spain, cultivated by the traditional method. RESULTS: PAL specific activity was assayed and hyperbolic kinetics were observed in both organs. The K_m value for L ‐Phe was 0.22 mmol L^?1 in leaf and 0.26 mmol L^?1 in fruit. In leaf, the highest PAL specific activity was found in the stage prior to veraison. By immunoblot, a PAL‐immunoreactive 75 kDa polypeptide was detected in leaf and fruit. In leaf, the level of this protein progressively rose until the last stages of ripening at the same time that total phenols increased. In fruit, PAL activity and protein change as in two series coinciding with different fruit‐maturation period. By immunohistochemistry under light microscopy, PAL was located in the epidermis and parenchyma cells of leaf and fruit. CONCLUSION: These results demonstrate the involvement and regulation of PAL during fruit ripening of olive, cv. Picual. Copyright © 2008 Society of Chemical Industry     

Human absorption and metabolism of oleuropein and hydroxytyrosol ingested as olive (Olea europaea L.) leaf extract

Phenolic compounds derived from the olive plant (Olea europaea L.), particularly hydroxytyrosol and oleuropein, have many beneficial effects in vitro. Olive leaves are the richest source of olive phenolic compounds, and olive leaf extract (OLE) is now a popular nutraceutical taken either as liquid or capsules. To quantify the bioavailability and metabolism of oleuropein and hydroxytyrosol when taken as OLE, nine volunteers (five males) aged 42.8 ± 7.4 years were randomized to receive either capsulated or liquid OLE as a single lower (51.1 mg oleuropein, 9.7 mg hydroxytyrosol) or higher (76.6 mg oleuropein, 14.5 mg hydroxytyrosol) dose, and then the opposite strength (but same formulation) a week later. Plasma and urine samples were collected at fixed intervals for 24 h post‐ingestion. Phenolic content was analyzed by LC‐ESI‐MS/MS. Conjugated metabolites of hydroxytyrosol were the primary metabolites recovered in plasma and urine after OLE ingestion. Peak oleuropein concentrations in plasma were greater following ingestion of liquid than capsule preparations (0.47 versus 2.74 ng/mL; p = 0.004), but no such effect was observed for peak concentrations of conjugated (sulfated and glucuronidated) hydroxytyrosol (p = 0.94). However, the latter peak was reached earlier with liquid preparation (93 versus 64 min; p = 0.031). There was a gender effect on the bioavailability of phenolic compounds, with males displaying greater plasma area under the curve for conjugated hydroxytyrosol (11 600 versus 2550 ng/mL; p = 0.048). All conjugated hydroxytyrosol metabolites were recovered in the urine within 8 h. There was wide inter‐individual variation. OLE effectively delivers oleuropein and hydroxytrosol metabolites to plasma in humans.     

LLE-UPLC-FLD法测定橄榄油中羟基酪醇和酪醇的含量

建立了液液萃取-超高效液相色谱串联荧光检测器(LLE-UPLC-FLD)快速分离和测定橄榄油中羟基酪醇和酪醇含量的方法。橄榄油样品经乙醇提取后,以0.2%(v/v)甲酸乙腈溶液和0.2%(v/v)甲酸水为流动相,梯度洗脱,采用BEH(50 mm×1.0 mm,1.7 μm)色谱柱,柱温为35 ℃,流速为0.15 mL/min,进样量为2.0 μL,使用荧光检测器在激发波长为300 nm,发射波长为350 nm下进行检测,整个分析过程仅需10 min。羟基酪醇和酪醇的检出限(S/N=3)及定量限(S/N=10)分别为0.11,0.13和0.31,0.39 μg/g,2种组分按高、中、低3个浓度水平加标回收率均高于89.52%;在最优条件下,对20批不同产地橄榄油中的羟基酪醇和酪醇进行检测,羟基酪醇和酪醇的含量范围分别在1.28~20.34及1.03~21.22 μg/g之间;该法简便快速,重复性良好,结果准确可靠,可用于橄榄油中酪醇和羟基酪醇含量的测定。     

Direct analysis of glucuronidated metabolites of main olive oil phenols in human urine after dietary consumption of virgin olive oil

In the present study we report on a UPLC-MRM validated method for the simultaneous direct analysis of main glucuronidated metabolites of olive oil phenols: tyrosol, hydroxytyrosol and its O-methyl metabolite homovanillyl alcohol in human urine after dietary olive oil ingestion. The developed method was linear within the concentration range 20–2000 ng/mL with adequate recovery of analytes (>86%). Intra- and inter-day precision and accuracy were according to standard requirements for method validation criteria. Using the developed method, urinary concentrations and excretion rates of glucuronides of olive oil phenols were successfully estimated in an intervention study with 11 healthy volunteers supplemented with a dietary dose of virgin olive oil (VOO) (50 mL). Therefore, about 13% of the consumed olive oil polyphenols were recovered in 24-h urine, where 75% of them were in the form of glucuronides (3′- and 4′-O-hydroxytyrosol glucuronides, 4′-O-glucuronides of tyrosol and homovanillyl alcohol) and 25% as free compounds.     

Impact of olive oil phenolic concentration on human plasmatic phenolic metabolites

Three different functional phenol-enriched virgin olive oils (FVOO) were prepared with a phenolic content of 250 (L-FVOO), 500 (M-FVOO), and 750 mg (H-FVOO) total phenols/kg. In a randomised, cross-over study with 12 healthy volunteers, the pharmacokinetics of phenolic biological metabolites was assessed. An increasing linear trend was observed for hydroxytyrosol sulfate, the main phenolic metabolite quantified in plasma, with C_max values of 1.35, 3.32, and 4.09 μmol/l, and AUC mean values of 263.7, 581.4, and 724.4 μmol/min for L-FVOO, M-FVOO, and H-FVOO, respectively. From our data an acute intake of phenol-enriched olive oils promotes a dose-dependent response of phenol conjugate metabolites in human plasma. Also, we point out for the first time hydroxytyrosol acetate sulfate as a main biological metabolite of hydroxytyrosol from olive oil ingestion.     

Activity of plant extracts for preserving functional food containing<Emphasis Type="Italic"> n</Emphasis>-3-PUFA

The ability of antioxidants obtained from natural sources to stabilise foodstuffs containing long n-3 fatty acids of marine origin has been determined. Food systems enriched in polyunsaturated fatty acids (PUFA) were: emulsified horse mackerel (Trauchurus trauchurus) muscle, fish oil-in-water emulsions (4% of n-3 PUFA) and fish oils (40% of n-3 PUFA). Rosemary leaves and extra virgin olive oil (EVO) were employed as sources of natural phenolic antioxidants. Both vegetable extracts were able to retard lipid oxidation in the different lipid systems. Rosemary extracts with a high content of carnosoic acid showed a significant synergism with fish proteins, by reinforcing their antioxidative effectiveness. Fish proteins and EVO-phenolics showed minor cooperative effects for inhibiting oxidation. The antioxidative partition into the different phases in the emulsified systems showed minor amounts of phenolics in the aqueous phases, except hydroxytyrosol and tyrosol, and a high adsorption on fish muscle.An erratum to this article can be found at     

Effect of phospholipids on extraction of hydrophilic phenols from virgin olive oils

Three methods, most frequently used in hydrophilic phenols extraction from virgin olive oils (liquid–liquid, solid-phase with C18 and solid phase with diol-bound sorbents), were applied on virgin olive oils enriched with 2.0–10.0 mg/kg of phospholipids (granular de-oiled soy lecithin). Phospholipids addition significantly decreased the total phenols concentration determined colorimetrically. Liquid–liquid extraction showed the best repeatability and recovery, and the lowest decrease of total phenols extracted in the presence of phospholipids. Extraction rate declined with the increase of phospholipids concentration, but showed a kind of saturation behaviour. Addition of phospholipids (5 g/kg) to various commercial extra virgin olive oils resulted in a different degree of total phenols decrease (from 1% to 45%), probably because of different affinity of phospholipids toward different classes of phenolic compounds. During prolonged contact time between phenols and phospholipids (60-days storage), oxidation monitored by K270 and a decrease of total phenols concentration proceeded more rapidly in the presence of phospholipids.     

多花黄精非药用部位活性成分及抗氧化性比较

目的:提高多花黄精的综合利用。方法:测定多花黄精根、茎、叶、花和嫩芽5个非药用部位的总黄酮、总皂苷和总酚含量,并比较非药用部位提取液的DPPH·、·OH及对亚硝酸盐的清除率。结果:多花黄精5个部位中总黄酮和总酚含量顺序大小一致,依次为叶>嫩芽>花>茎>根,总皂苷以花中含量最高;抗氧化活性方面,叶提取液的DPPH·、·OH和对亚硝酸盐的清除率最高,其次是嫩芽、花。相关性分析发现,多花黄精非药用部位提取液对DPPH·、·OH和亚硝酸盐的清除率与总黄酮、总皂苷和总酚含量均表现出一定的正相关。结论:多花黄精非药用部位提取液具有较好的生物活性。     

Emerging Technologies for Recovery of Value-Added Components from Olive Leaves and Their Applications in Food/Feed Industries

Olive leaves are the most abundant agricultural waste source rich in polyphenolics. Due to the numerous health benefits associated with these compounds, the interest in recovering polyphenols from olive leaves has increased in the scientific community over the last decade. Recent studies have focused on improved extraction techniques and processing methods that are most suited for agro-biological industries involved in the development of nutraceutical and functional products. The major problems in olive leaves processing include bitter taste and the low stability of various phenolic compounds. Oleuropein and hydroxytyrosol are the most important phenolic compounds extracted from olive leaves. The present review highlights the importance of olive leaves, their composition, preparation methods, major phenolic compounds, and commercial applications. This review article focuses on integrating studies on olive leaf extract (OLE) pertinent to nutrition, health, and beauty. The different board categories of delivery systems available for the encapsulation of OLE are given. These novel delivery systems could improve fortification, supplementation, and dietary diversification in food and pharmaceutical products.     

Identification of a new phenolic compound from Chinese olive (<Emphasis Type="Italic">Canarium album</Emphasis> L.) fruit

Chinese olive (Canarium album L.), a native and a well-known tropical fruit tree in the southeast of China, contains large amount of phenolics and possesses great pharmacological activities. In this study, phenolics were extracted from Chinese olive fruit pulp using 80% (v/v) aqueous acetone, and acetone extracts were further fractioned with petroleum ether, ethyl acetate and n-butanol sequentially. From n-butanol fraction, a new phenolic compound was isolated and purified through AB-8 adsorption resin column chromatography, polyamide column chromatography and TSK Toyopearl HW-40 (S) column chromatography, and the structure of the new compound was established as 3-O-galloyl quinic acid butyl ester by electrospray ionization mass spectrometry (ESI-MS), 1D- and 2D-NMR (DEPT, COSY, HMBC, HMQC) and UV–vis techniques.     

Effect of olive ripeness on chemical properties and phenolic composition of chétoui virgin olive oil

BACKGROUND: The aim of the present work was to investigate the influence of fruit ripening on oil quality in an attempt to establish an optimum harvesting time for Chétoui olives, the second main olive variety cultivated in Tunisia. RESULTS: Our results showed that many analytical parameters, i.e., peroxide value, UV absorbance at 232 and 270 nm, chlorophyll pigments, carotenoids and oleic acid contents decreased during ripening, whilst oil content and linolenic acid increased. Free acidity remained practically stable with a very slight rise at the highest maturity index. The trend of oxidative stability, total phenols and o‐diphenols, showed an increase at the early stages followed by a reduction at more advanced stages of maturity. The major phenolic compounds, such as hydroxytyrosol, ligstroside aglycon, elenolic acid, acetoxy‐pinoresinol and oleuropein aglycon, seemed to have the same behaviour. In the case of tyrosol, a strong decrease was observed directly related with the ripening progress. CONLUSION: On the basis of the evolution of the analytical parameters studied, the best stage of Chétoui olive fruits for oil processing seems to be at ripeness index higher than 2.0 and lower than 3.0. Copyright © 2009 Society of Chemical Industry     

Transfer of phenolic compounds during olive oil extraction in relation to ripening stage of the fruit

The transfer of phenolic compounds of Olea europaea L. cv. Arbequina variety during olive oil extraction in relation to ripening stage was investigated. The parameters of oil extraction by the Abencor system are shown together with mass balances of the products and by products from the olive oil extraction in relation to olive paste. The phenolic compounds in olive paste, pomace, oil and wastewater were identified and measured by HPLC. Throughout the study, the concentrations of simple phenols, secoiridoids and flavonoids were higher in the olive paste and pomace phases than in oil and wastewater phases. High concentrations of 4‐(acetoxyethyl)‐1,2‐dihydroxybenzene (3,4‐DHPEA‐AC) and secoiridoid derivatives such as the dialdehydic form of elenolic acid linked to 3,4‐DHPEA (hydroxytyrosol) or p‐HPEA (tyrosol) (3,4‐DHPEA–EDA, p‐HPEA–EDA, where EDA is elenolic acid dialdehyde) and an isomer of oleuropein aglycone (3,4‐DHPEA–EA, where EA is elenolic acid aldehyde) were found in olive oil, together with lignan compounds. It was observed that 3,4‐DHPEA–EDA was the most abundant polyphenol present in the wastewater phase. This indicates that biotransformation occurred during olive extraction, especially in the crushing and malaxation operations, and reflects the possible chemical changes that lead to the formation of new compounds. Moreover, the distribution of compounds showed their affinities toward different phases. Copyright © 2005 Society of Chemical Industry