Biofilm Inhibition of Inula viscosa (L.) Aiton and Globularia alypum L. Extracts Against Candida Infectious Pathogens and In Vivo Action on Galleria mellonella Model.

The increasing importance of fungal infections has fueled the search for new beneficial alternatives substance from plant extracts. The current study investigates the antifungal and antibiofilm activity of Inula viscosa (L.) Aiton and Globularia alypum (L.) leaves extracts against Candida both in vitro and in vivo. The inhibition of planktonic and sessile Candida albicans and Candida glabrata growth using both leaf extracts are evaluated. Moreover; an in vivo infection model using Galleria mellonella larvae; infected and treated with the extracts are performed. All extracts show fungicidal activity; with a minimum fungicidal concentration (MFC) ranging from 128 to 512 µg mL-1 against the two selected strains of Candida. In particular, the best results are obtained with methanolic extract of I. viscosa and G. alypum with an MFC value of 128 µg mL-1 . The extracts are capable to prevent 90% of biofilm development at minor concentrations ranging from 100.71 ± 2.49 µg mL-1 to 380.4 ± 0.92 µg mL-1 . In vivo, tests on Galleria mellonella larvae show that the extracts increase the survival of the larvae infected with Candida. The attained results reveal that I. viscosa and G. alypum extracts may be considered as new antifungal agents and biofilm inhibiting agents for the pharmaceutical and agro-food field.

Comparison between Ultrasound- and Microwave-Assisted Extraction Methods to Determine Phenolic Compounds in Barley (Hordeum vulgare L.).

Barley (Hordeum vulgare L.) is one of the major cereal crops worldwide. It is grown not only to be used as fodder but also for human consumption. Barley grains are a great source of phenolic compounds, which are particularly interesting for their health-promoting antioxidant properties, among other benefits. Two extraction methods, namely ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE), have been optimized and compared by using Box-Behnken design (BBD) to determine both the antioxidant power and the phenolic compound levels of the extracts. Three variables have been assessed based on these designs: solvent composition (% MeOH in water), temperature (°C), and sample-to-solvent ratio (mg sample mL-1 solvent). The solvent composition used and the interaction between the solvent and the temperature were the most significant variables in terms of recovery of phenolic compounds and antioxidant capacity of the extracts. Short extraction times, a high precision level, and good recoveries have been confirmed for both methods. Moreover, they were successfully applied to several samples. Significant differences regarding the level of phenolic compounds and antioxidant power were revealed when analyzing three different barley varieties. Specifically, the amounts of phenolic compounds ranged from 1.08 to 1.81 mg gallic acid equivalent g-1 barley, while their antioxidant capacity ranged from 1.35 to 2.06 mg Trolox equivalent g-1 barley, depending on the barley variety. Finally, MAE was found to be slightly more efficient than UAE, presenting higher levels of phenolic compounds in the extracts.

Evaluation of Different Extraction Methods on the Phenolic Profile and the Antioxidant Potential of Ceratonia siliqua L. Pods Extracts.

The present work was designed to investigate the effects of different extraction processes, namely ultrasonic-assisted, supercritical fluid, microwave-assisted and Soxhlet applied to carob pods. The total phenolic quantification and the antioxidant activity were assessed by the means of rapid in vitro spectrophotometric assays; the phenolic profile was identified using ultra-high performance liquid chromatography coupled to mass spectrometry. The results revealed that the phenolic compounds and the antioxidant capacity varied significantly with the nature of the extraction process. The content of total phenolic compounds ranged from 11.55 to 34.38 mg GAE/g DW; the content of total flavonoids varied from 3.50 to 10.53 mg QE/g DW, and the content of condensed tannins fluctuated from 3.30 to 6.55 mg CE/ g DW. All extracts performed differently on antioxidant activity when determined by the DPPH assay producing a dose-dependent response, with IC50 extended from 11.33 to 6.07 µg/mL. HPLC analysis enabled the identification of nine compounds. As a function of the studied extraction methods, the phenolic compound contents were positively correlated with antioxidant activity.

Elucidation of Antioxidant Compounds in Moroccan Chamaerops humilis L. Fruits by GC-MS and HPLC-MS Techniques.

The aim of this study was to characterize the phytochemical content as well as the antioxidant ability of the Moroccan species Chamaerops humilis L. Besides crude ethanolic extract, two extracts obtained by sonication using two solvents with increased polarity, namely ethyl acetate (EtOAc) and methanol-water (MeOH-H2O) 80:20 (v/v), were investigated by both spectroscopy and chromatography methods. Between the two extracts, the MeOH-H2O one showed the highest total polyphenolic content equal to 32.7 ± 0.1 mg GAE/g DM with respect to the EtOAc extract (3.6 ± 0.5 mg GAE/g DM). Concerning the antioxidant activity of the two extracts, the EtOAc one yielded the highest value (1.9 ± 0.1 mg/mL) with respect to MeOH-H2O (0.4 ± 0.1 mg/mL). The C. humilisn-hexane fraction, analyzed by GC-MS, exhibited 69 compounds belonging to different chemical classes, with n-Hexadecanoic acid as a major compound (21.75%), whereas the polyphenolic profile, elucidated by HPLC-PDA/MS, led to the identification of a total of sixteen and thirteen different compounds in both EtOAc (major component: ferulic acid: 104.7 ± 2.52 µg/g) and MeOH-H2O extracts (major component: chlorogenic acid: 45.4 ± 1.59 µg/g), respectively. The attained results clearly highlight the potential of C. humilis as an important source of bioactive components, making it a valuable candidate to be advantageously added to the daily diet. Furthermore, this study provides the scientific basis for the exploitation of the Doum in the food, pharmaceutical and nutraceutical industries.

Physico-Chemical and Phytochemical Characterization of Moroccan Wild Jujube "Zizyphus lotus (L.)" Fruit Crude Extract and Fractions.

Wild jujube "Ziziphus lotus (L.) Desf." belongs to the Rhamnaceae family and is a traditionally herbaceous medicinal plant. It is very common in arid and semi-arid regions and is currently used for its antidiabetic, sedative, analgesic, anti-inflammatory and hypoglycemic activities. The aim of the present work was to characterize the physico-chemical properties and the phytochemical profile of wild jujube sample collected from the Guercif region, in order to determine the polyphenolic compounds and the antioxidant ability Analyses were carried out directly after the harvest for the determination of pH, refractive index, total soluble solid (°Brix), dry matter, sugar/acidity, total sugars, reducing sugars, as well as lipid and protein content. Results showed that the investigated fruit is acidic (pH 4.9 ± 0.23) and rich in sugars (80.2 g/100 g ± 3.81). The GC-MS analysis of the fruit revealed a number of volatile compounds, as many as 97, belonging to different chemical classes. The HPLC-DAD-ESI/MS analysis showed the presence of a total of 20 polyphenolic compounds in both EtOAc and MeOH-water extracts. Among them, p-Hydroxybenzoic acid was the most abundant in the EtOAc extract (185.68 µg/100 mg ± 0.5) whereas Quercetin 3-O-rhamnoside-7-O-glucoside was found in higher amounts in the MeOH-water extract (25.40 µg/100 mg ± 0.5). These components have medical interest, notably for human nutrition, as well as health benefits and therapeutic effects. Therefore, Moroccan jujube "Zizyphus lotus (L.)" fruit may have potential industrial applications for food formulations.

Investigation on phenolic compounds stability during microwave-assisted extraction.

The stability of 22 phenolic compounds of different families (benzoic acids, benzoic aldehydes, cinnamic acids, catechins, coumarins, stilbens and flavonols) has been studied under conditions of microwave-assisted extraction. The influence on the stability affected by the working temperature between 50 and 175 degrees C has been evaluated, and it has been concluded that all the compounds studied are stable up to 100 degrees C, whereas at 125 degrees C there is significant degradation of epicatechin, resveratrol and myricetin. Conclusions have been drawn on the relationship between the chemical structure and the stability of the compounds; it has been found that those that have a greater number of hydroxyl-type substituents are more easily degraded under the extraction conditions.

Investigation on Ochratoxin A stability using different extraction techniques.

The stability of Ochratoxin A during its extraction using different extraction techniques has been evaluated. Microwave-assisted extraction and pressurised liquid extraction, in addition to two other reference methods of extraction, i.e. ultrasound-assisted and magnetic stirring-assisted extraction, were evaluated. The effect of extraction temperature using the cited techniques was checked. The results show that Ochratoxin A can be extracted using microwave-assisted extraction at temperatures up to 150 degrees C without degradation. Pressurised liquid extraction can be used at temperatures up to 100 degrees C, for extraction times of less than 30min. Further, both ultrasound-assisted extraction and magnetic stirring extraction can be applied at temperatures up to 65 degrees C. High-performance liquid chromatography combined with fluorescence detection using a Chromolith RP-18e column at a flow rate of 5mL min(-1) was used to quantify the Ochratoxin A. The retention time for the Ochratoxin A was 1.3min. The limits of detection (LOD) and of quantification (LOQ) were 0.03 and 0.10microg L(-1), respectively.