Ultrasound-assisted extraction, optimisation using response surface methodology and HPLC-DAD phenolic compounds quantification from Passiflora edulis S. peels cultivated in Tunisia.

A new ultrasound-assisted extraction method was developed for the determination of phenolic compounds extracted from Passiflora edulis Sims peels cultivated in Tunisia. Several extraction variables including: extraction time, temperature, liquid/solid ratio and pH have been studied using response surface methodology. The extraction efficiency was evaluated by measuring the total phenolics; flavonoids and antioxidants content. The highest values of the studied response were observed after 9.78 min at 49.64 °C in a liquid-to-solid ratio 22.07 ml/g and in pH (5.54). The individual phenolic compounds content of the optimum extract peels of the Passiflora edulis Sims have been analysed using HPLC-DAD. The results revealed the presence of gallic followed by caffeic acids, while the apigenin-7-glucoside and rutin have been quantified as the abundant flavonoid compounds (0.653 and 0.488 mg/ml, respectively). This green procedure should be a promising option to guide industrial design for the production of phenolic-rich plant extracts.

Chemical Composition and Bioactivities of Quercus canariensis Flour Acorns Extracts Growing in Tunisia.

The current study was conducted to investigate the chemical composition of Quercus canariensis flour acorns extracts as well as its biological activities in regards to the growing area using spectrophotometric and chromatographic techniques. The phenolic profile was composed of 19 compounds identified through HPLC-DAD analysis. Coumarin was the most abundant compound quantified in BniMtir, Nefza and ElGhorra and gallic (12.58-20.52 %), syringic (4.70-7.64 %) and trans-ferulic (2.28-2.94 %) acids were the abundant phenolic acids while kaempferol was the major flavonoid compounds quantified only in Quercus canariensis growing in BniMtir. On the other hand, Ain Snoussi acorn extract was characterized by its high content in luteolin-7-O-glucoside (58.46 %). The in-vitro antioxidant activities of the studied extracts were investigated and the results showed that Nefza ethanolic extract's has the highest activities. A bactericidal effect against Staphylococcus aureus was observed only by Elghorra population. On the other hand, Ain Snoussi acorn extract was efficient to inhibit growth of pathogenic bacteria, mentioned the highest activity against Escherichia coli. These results is the first study highlighted that zeen oak acorns are an excellent source of natural antioxidants and antibacterial compounds related to their lysozyme activity which could be exploited in the pharmaceutical and food sectors.

Antibacterial, antibiofilm and cytotoxic properties of prodigiosin produced by a newly isolated Serratia sp. C6LB from a milk collection center.

The aim of this work was to produce a red tripyrrole pigment prodigiosin from Serratia sp. C6LB strain, to investigate the promising antimicrobial properties on Gram-positive and Gram-negative bacterial strains. The research was also proposed to evaluate the antibiofilm activity on Staphylococcus epidermidis S61 biofilm and its cytotoxic activity against human cancer cell lines. The production and structural elucidation of prodigiosin was carried out using spectrophotometric scanning, TLC, HPLC, FTIR and NMR analysis. The pigment production was optimized using mannose and peptone as carbon and nitrogen sources, respectively. The study confirmed promising antibacterial properties of prodigiosin on eight Gram-positive and Gram-negative bacterial strains with MICs values ranged from 0.039 to 2.5 mg/mL. Antiadhesive activity test of prodigiosin on Staphylococcus epidermidis S61 biofilm exhibited 99.9% inhibition, whereas maximum biofilm eradication activity reached 65%. Cytotoxic activity showed IC50 of 16 µg/mL and 6.7 µg/mL against breast cancer lines MCF-7 and MDA-MB231, respectively.

Tunisian Native Mentha pulegium L. Extracts: Phytochemical Composition and Biological Activities.

Mint species (Lamiaceae family) have been used as traditional remedies for the treatment of several diseases. In this work, we aimed to characterize the biological activities of the total phenolic and flavonoid contents of Mentha pulegium L. extracts collected from two different regions of Tunisia. The highest amounts of total phenols (74.45 ± 0.01 mg GAE/g DW), flavonoids (28.87 ± 0.02 mg RE/g DW), and condensed tannins (4.35 ± 0.02 mg CE/g DW) were found in the Bizerte locality. Methanolic leaf extracts were subjected to HPLC-UV analysis in order to identify and quantify the phenolic composition. This technique allowed us to identify seven phenolic compounds: two phenolic acids and five flavonoid compounds, such as eriocitrin, hesperidin, narirutin, luteolin, and isorhoifolin, which were found in both extracts with significant differences between samples collected from the different regions (p < 0.05). Furthermore, our results showed that the methanolic extract from leaves collected from Bizerte had the highest antioxidant activities (DPPH IC50 value of 16.31 µg/mL and 570.08 µmol Fe2+/g, respectively). Both extracts showed high radical-scavenging activity as well as significant antimicrobial activity against eight tested bacteria. The highest antimicrobial activities were observed against Gram-positive bacteria with inhibition zone diameters and MIC values ranging between 19 and 32 mm and 40 and 160 µg/mL, respectively. Interestingly, at 10 µg/mL, the extract had a significant effect on cell proliferation of U87 human glioblastoma cells. These findings open perspectives for the use of Mentha pulegium L. extract in green pharmacy, alternative/complementary medicine, and natural preventive therapies for the development of effective antioxidant, antibacterial, and/or antitumoral drugs.

Identification and quantification of phenolic and fatty acid profiles in Pinus halepensis mill. seeds by LC-ESI-MS and GC: Effect of drying methods on chemical composition.

This study aimed to evaluate Pinus halepensis Mill. seeds oil as well as methanolic-aqueous extract on fatty acid and phenolic's composition as well as antioxidant activities with regard to the cones drying methods: convection and sun methods. The highest amounts of total phenols (14.63 ± 0.05 mg GAE/g DW), flavonoids (3.3 ± 0.02 mg QE/g DW), and condensed tannins (0.36 ± 0.05 mg CE/g DW) were showed in the seeds obtained by sun-dried method. Methanolic-aqueous seeds extracts were subjected to LC-ESI-MS analysis in order to identify and quantify the phenolic composition. This technique allowed us to identify eleven phenolic compounds: two phenolic acids and nine flavonoid compounds such as cirsiliol, catechin (+), luteolin, and luteolin-7-O-glucoside, which were present in the two studied samples while apigenin, naringenin, and cirsilineol were only identified in the extract obtained from sun-drying method seeds. The chemical components of the oils were analyzed using GC, and significant differences were found between the two studied seeds oil (p < .05). Furthermore, the antioxidant activities were investigated using DPPH and ABTS.+ assays. The results showed that the methanolic-aqueous extract from seeds dried by sun method had the highest antioxidant activities (0.08 and 0.05 mg/ml, respectively). This study could provide useful information for industry to produce potentially bioactive plant extract.

Pistacia atlantica Desf. roots extract: LC-ESI-MS Analysis, antioxidant activity and gastroprotective effect on experimentally-induced ultrastructural gastric ulcers in mice.

Pistacia atlantica Desf. (Atlas Pistachio) is an Anacardiaceae tree traditionally used in Tunisia for the treatment of ophthalmic, stomatitis, and digestive tract diseases. In the present study, the Pistacia atlantica Desf. roots extract (PR) was phytochemically analyzed, for the first time, by LC-ESI-MS for phenolic and flavonoid contents, in vitro tested for its potential antioxidant activity based on the 2.2-diphenyl-1-picrylhydrazyl (DPPH) and the reduced power essays (FRAP), and in vivo tested for its ability to shield against ethanol-induced gastric ulcer in mice. The LC-ESI-MS analysis proved the identification of 12 compounds, including Quinic, Gallic, and Protocatechuic, as major phenolic acids and high levels of flavonoids, such as Catechin, Epicatechin, and Cirsiliol. PR also exhibited a mild in vitro antioxidant activity when compared with ascorbic acid. In vivo pretreatment of ethanol-ulcerated mice with PR doses 50 mg/kg and 100 mg/kg body weight (b.w) significantly reduced (P< .05) gastric lesions at a rate of 20.10% and a rate of 40.90%, respectively, when compared with 60.70% rate of sucralfate (50 mg/kg b.w) evidenced by a dose-dependent manner increase in the gastric mucosa enzymatic (SOD, CAT, GPx) antioxidant levels, the decline of the lipid peroxidation, and the preservation of normal gastric superficial epithelium. The underlying mechanism of PR antiulcerogenic activity could be due to a synergistic effect of phenolic acids and flavonoid contents which enhances the gastric antioxidant defense system.Abbreviations: BHT: butylated hydroxytoluene, b.w: body weight, CAT: catalase, DPPH:1-Diphenyl-2-picrylhydrazyl, DW: dry weight, EtOH: ethanol, FRAP: Ferric reducing antioxidant power, GAE: gallic acid equivalents, GPx: Glutathione peroxidase, QE: quercetin equivalents, LC-ESI-MS: Liquid chromatography-Electrospray Ionization-Tandem Mass Spectrometry, MDA: malondialdehyde, PR: Pistacia root, TBA: thiobarbituric acid reagent, TBARS: thiobarbituric acid reactive substances, TCA: trichloroacetic acid, SOD: Superoxide dismutase.

Phenolic profile (HPLC-UV) analysis and biological activities of two organic extracts from Echinops spinosissimus Turra roots growing in Tunisia.

Echinops spinosissimus Turra subsp. roots was investigated, after extraction with two different solvents: ethanol and ethyl acetate, for its phenolic profile as well as biological activities. Echinops spinosissimus Turra subsp. spinosus roots ethanolic extract was characterized by the highest content of total polyphenols (19.3 mg GAE/100 g of DW). In addition, the phenolic profiles of the two studied extracts were analysed using HPLC-UV, where p-coumaric (8.59 mg/kg of DW) and cinnamic acids (4.68 mg/kg of DW) were detected as the most abundant phenolic acids, and kaempferol was the predominant flavonoids (30.37 mg/kg of DM) followed by quercetin and rutin in the ethanolic extract. While, only p-coumaric acid, epicatechin, kaempferol and quercetin were identified and quantified in ethyl acetate roots extract. All tested extracts exhibited an antimicrobial effect against Gram-positive and Gram-negative bacteria. These findings indicate that Echinops spinosissimus Turra subsp. spinosus roots can be regarded as a new source of edible oil having health benefits and nutritional properties.

Change in some quality parameters and oxidative stability of olive oils with regard to ultrasound pretreatment, depitting and water addition.

Ultrasound pretreatment with different times (0, 4, 8, 10 min) on olive paste previous malaxation of olive oil extraction along with depitting and water application procedures were studied. The effects of these procedures on oil yield, quality parameters and oxidative stability have been studied. Therefore, the olive oil yield increase with increasing time of ultrasound treatment. On the other hand, the application of ultrasound did not adversely affect the quality characteristics, as well as the antioxidant activity, when comparing with untreated paste. Furthermore, the oxidative stability data, we can conclude that ultrasound treatment can affect the olive oil oxidative stability. This study could provide useful information for industry to produce olive oil with high yield and quality.

Phenolic content, antioxidant capacity and antimicrobial activity of leaf extracts from Pistacia atlantica.

The aim of this study was to investigate the secondary metabolites related to Pistacia atlantica Desf as well as their biological activities. Results show that using ethanol as a solvent exhibited the highest content of total phenolics and flavonoids (68.23 mg GAE g-1 of DW and 44 mg RE g-1 of DW), followed by aqueous one (20.07 mg GAE g-1 of DW and 15 mg RE g-1 of DW). Good correlation (Data not shown) was obtained between the DPPH radical-scavenging activities and total phenolics contents. Many similarities were observed between the results of the DPPH (IC50 = 32 and 200 µg mL-1) and ABTS (IC50 = 42 and 300 µg mL-1) assays. All tested extracts contained phenolic compounds exhibited an antimicrobial effect against Gram-positive and Gram-negative bacteria. Further, researches for identification and purification of phenolic compounds are required.

Effect of storage on refined olive oil composition: stabilization by addition of chlorophyll pigments and squalene.

Refined olive oil has been analyzed in order to evaluate the influence of storage time on their quality after enrichment with chlorophyll pigments and squalene. At the end of the storage, chlorophyll pigments decomposed for more than 90%, while, carotene pigment loss was lower showing up to 75% in all oil samples stored at 20°C and 50°C. The reduction of total phenolic compounds exhibited similar degradation profiles, reducing by 81% for enriched refined olive oil stored at 20°C and 50°C after six months. For all studied samples, squalene content decreased significantly (p< 0.05) only after four months of storage. Oils with added pure squalene had the lower peroxide values (52 meq O2 · kg(-1)) at the end of storage at 20°C. After six months of storage the oxidative stability increased to 1 h 28 min and 2 h 06 min for refined olive oil stored at 20°C enriched with chlorophyll + squalene and only by squalene, respectively. Therefore, squalene supplemented with 800 mg · kg(-1) exhibited favorable antioxidant effects and is preferable for effectively avoiding oxidation. This study could provide useful information for industry to produce high-stability vegetable oil.

Characterisation and phenolic profiles of two rare olive oils from southern Tunisia: Dhokar and Gemri-Dhokar cultivars.

BACKGROUND: The aim of this work was to study the chemical characteristics of two Tunisian cultivars, namely Dhokar and Gemri-Dhokar, to analyse the fatty acids, sterols, triacylglycerols, triterpenic alcohols, and to determine the phenolic composition and oxidative stability. RESULTS: Among the rare varieties, Gemri-Dhokar olive oil had the highest value of oleic acid (69.39%) whereas Dhokar oil was noteworthy for its lower content of phenolic compounds (94.56 mg kg(-1) gallic acid equivalents of oil) and presented the highest level of palmitic acid (19.37%). The main sterols found in all olive oil samples were ß-sitosterol and Δ5-avenasterol, whereas cholesterol and 24-methylenecholesterol were also found in all samples but in lower amounts. Two triterpenic dialcohols (erythrodiol and uvaol) were also detected and their content ranged from 1.45 to 2.30%, in Gemri-Dhokar and Dhokar olive oil, respectively. Ten phenolic compounds were identified. In all samples, the main phenols found were oleuropein aglycon and pinoresinol. These phenolic compounds showed significant correlations with oxidative stability. CONCLUSION: The analytical parameters of two oils that were determined in this study were greatly influenced by genetic factors (cultivar).

Analytical evaluation of two monovarietal virgin olive oils cultivated in the south of Tunisia: Jemri-Bouchouka and Chemlali-Tataouin cultivars.

BACKGROUND: The characterisation of virgin olive oils from two Tunisian cultivars, growing in the Tataouin zone, namely Jemri-Bouchouka, a rare olive cultivar, and Chemlali-Tataouin, was carried out. Several analytical parameters were evaluated; these include quality index, fatty acids, phenolic, chlorophyll, carotenoid, squalene, α-tocopherol compositions and oxidative stability. RESULTS: Jemri-Bouchouka olive oil had the highest value of oleic acid (74.50%) while Chemlali-Tataouin olive oil had the highest value of oleic acid (69.39 %) and also was characterized by a high percentage of palmitic acid (14.75 %) which makes this oil freeze at a low temperature [corrected]. On the other hand, Jemri-Bouchouka oil was characterised by a low phenolic and α-tocopherol content (267.72 mg GAE kg⁻¹ and 278.34 mg kg⁻¹, respectively). Ten phenolic compounds were identified. The main phenols found in the two olive oils were oleuropein aglycon and pinoresinol. All phenolic compounds showed significant correlations with oxidative stability. CONCLUSION: The analytical parameters of virgin olive oil that were determined in this study were greatly influenced by cultivar.

Recovery of hydroxytyrosol rich extract from two-phase Chemlali olive pomace by chemical treatment.

UNLABELLED: A very simple method is proposed to produce hydroxytyrosol, a commercially unavailable compound with well-known biological properties which justify a potential commercial application. The 2-phase Chemlali olive pomace is selected as substrate for chemical treatment. Different conditions of chemical treatment, including concentration of acid and alkaline solutions, time and temperature, were assayed. A high amount of hydroxytyrosol (1360 mg/kg of fresh 2-phase olive pomace) was obtained using water bath after treatment at 80 °C for 90 min with 1 M of H(3) PO(4) . However, treatment of 2-phase Chemlali olive pomace using autoclave apparatus could produce a large amount of hydroxytyrosol (1993.60 and 1515.88 mg/kg of fresh alperujo, 1 M acid and basic catalyst, respectively). By taking into consideration practical and economic aspects, acid-catalyzed treatment was more effective using autoclave conditions, whereas the alkali catalyzed conditions were not very suitable. This study could provide useful information for industry to produce the potentially bioactive compound. PRACTICAL APPLICATION: The 2-phase Chemlali olive pomace is selected as substrate for chemical treatment. Treatment of "alperujo" using water bath or autoclave apparatus was carried out. A high amount of hydroxytyrosol was obtained using autoclave apparatus.

Phenolic composition, isolation, and structure of a new deoxyloganic acid derivative from Dhokar and Gemri-Dhokar olive cultivars.

Two rare olive cultivars, Dhokar and Gemri-Dhokar, growing in the south of Tunisia were investigated to identify their phenolic profile and evaluate their sugar content and antioxidant activity. The highest value of phenolic content was detected in Gemri-Dhokar cultivar extract (6.97 g gallic acid equivalents/kg of fresh olive). In addition, sugar content was quantified; glucose (45.17 g/kg of fresh olive) was the predominant sugar in Dhokar cultivar, followed by fructose (40.83 g/kg of fresh olive). The identification of phenolic compounds was based on separation by high-performance liquid chromatography equipped with a diode array detector followed by liquid chromatography-mass spectrometry analysis. In both cultivars, Oleuropein aglycon derivatives and elenolic acid were the main phenolic components. Oleuropeins were the major compounds quantified in the Gemri-Dhokar drupes olives (61.04 mg/100 g of fresh olive), while 0.25 mg/100 g were found in fresh Dhokar. A new iridoid compound, isolated as pure compound, was not previously reported in the literature. Its structure was established by spectroscopic analyses (NMR, UV, MS, and IR). DPPH, ABTS, and FRAP assays showed that the most important antioxidant capacity of olive extracts was found in with Gemri-Dhokar cultivar.