Chemical, Antioxidant and Biological Studies of Brassica incana subsp. raimondoi (Brassicaceae) Leaf Extract.

Brassica incana subsp. raimondoi is an endemic taxon present in a restricted area located on steep limestone cliffs at an altitude of about 500 m a.s.l. in eastern Sicily. In this research, for the first time, studies on the phytochemical profile, the antioxidant properties in cell-free and cell-based systems, the cytotoxicity on normal and cancer cells by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, and on Artemia salina Leach, were performed. The total phenolic, flavonoid, and condensed tannin contents of the leaf hydroalcoholic extract were spectrophotometrically determined. Ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) analysis highlighted the presence of several phenolic acids, flavonoids, and carotenoids, while High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) identified various kaempferol and isorhamnetin derivatives. The extract exhibited different antioxidant properties according to the five in vitro methods used. Cytotoxicity by MTT assay evidenced no impact on normal human fibroblasts (HFF-1) and prostate cancer cells (DU145), and cytotoxicity accompanied by necrotic cell death for colon cancer cells (CaCo-2) and hepatoma cells (HepG2), starting from 100 µg/mL and 500 µg/mL, respectively. No cytotoxic effects were detected by the A. salina lethality bioassay. In the H2O2-induced oxidative stress cell model, the extract counteracted cellular reactive oxygen species (ROS) production and preserved non-protein thiol groups (RSH) affected by H2O2 exposure in HepG2 cells. Results suggest the potential of B. incana subsp. raimondoi as a source of bioactive molecules.

Determination of the Phenolic Profile by Liquid Chromatography, Evaluation of Antioxidant Activity and Toxicity of Moroccan Erica multiflora, Erica scoparia, and Calluna vulgaris (Ericaceae).

This study aimed to investigate the phenolic profile and selected biological activities of the leaf and aerial extracts of three Ericaceae species, namely Erica multiflora, Erica scoparia, and Calluna vulgaris, collected from three different places in the north of Morocco. The phenolic composition of all extracts was determined by LC coupled with photodiode array and mass spectrometry detection. Among the investigated extracts, that of E. scoparia aerial parts was the richest one, with a total amount of polyphenols of 9528.93 mg/kg. Up to 59 phenolic compounds were detected: 52 were positively identified and 49 quantified-11 in C. vulgaris, 14 in E. multiflora, and 24 in E. scoparia. In terms of chemical classes, nine were phenolic acids and 43 were flavonoids, and among them, the majority belonged to the class of flavonols. The antioxidant activity of all extracts was investigated by three different in vitro methods, namely DPPH, reducing power, and Fe2+ chelating assays; E. scoparia aerial part extract was the most active, with an IC50 of 0.142 ± 0.014 mg/mL (DPPH test) and 1.898 ± 0.056 ASE/mL (reducing power assay). Further, all extracts were non-toxic against Artemia salina, thus indicating their potential safety. The findings attained in this work for such Moroccan Ericaceae species, never investigated so far, bring novelty to the field and show them to be valuable sources of phenolic compounds with interesting primary antioxidant properties.

Phenolic and Volatile Composition and Antioxidant Properties of the Leaf Extract of Brassica fruticulosa subsp. fruticulosa (Brassicaceae) Growing Wild in Sicily (Italy).

In continuation of research conducted on species of the spontaneous flora of Sicily (Italy) belonging to the Brassicaceae family, Brassica fruticulosa subsp. fruticulosa was selected. It is an edible species utilized in Sicilian traditional medicine. In this study, for the first time, the phenolic and the volatile compounds and the antioxidant properties of the hydroalcoholic extract obtained from the leaves of B. fruticulosa subsp. fruticulosa were characterized. Through HPLC-PDA/ESI-MS analysis, a total of 22 polyphenolic compounds (20 flavonoids and 2 phenolic acids) were identified, with 3-hydroxiferuloylsophoroside-7-O-glucoside (1.30 mg/g ± 0.01) and kaempferol-3-O-feruloylsophoroside-7-O-glucoside (1.28 mg/g ± 0.01) as the most abundant compounds. Through SPME-GC/MS several volatiles belonging to different chemical classes were characterized, with nitriles and aldehydes accounting for more than 54% of the whole volatile fraction. The extract of B. fruticulosa subsp. fruticulosa showed moderate activity in the DPPH assay (IC50 = 1.65 ± 0.08 mg/mL), weak reducing power (17.47 ± 0.65 ASE/mL), and good chelating properties (IC50 = 0.38 ± 0.02 mg/mL), reaching approximately 90% activity at the highest tested concentration. Lastly, the extract was non-toxic against Artemia salina, indicating its potential safety. According to the findings, it can be stated that B. fruticulosa subsp. fruticulosa represents a new valuable source of bioactive compounds.

Brassica incana Ten. (Brassicaceae): Phenolic Constituents, Antioxidant and Cytotoxic Properties of the Leaf and Flowering Top Extracts.

Brassica incana Ten. is an edible plant belonging to the Brassicaceae family. In this work, the phenolic composition and the antioxidant and cytotoxic properties of the hydroalcoholic extracts obtained from the leaves and the flowering tops of B. incana grown wild in Sicily (Italy) were studied for the first time. A total of 17 and 20 polyphenolic compounds were identified in the leaf and in the flowering top extracts, respectively, by HPLC-PDA-ESI-MS analysis. Brassica incana extracts showed in vitro antioxidant properties; the leaf extract displayed greater radical scavenging activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test than the flowering top extract (IC50 = 1.306 ± 0.049 mg/mL and 2.077 ± 0.011 mg/mL), which in turn had a stronger ferrous ion chelating ability than the other (IC50 = 0.232 ± 0.002 mg/mL and 1.147 ± 0.016 mg/mL). The cytotoxicity of the extracts against human colorectal adenocarcinoma (CaCo-2) and breast cancer (MCF-7) cell lines was evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the lactic dehydrogenase (LDH) release determination. The extracts showed cytotoxic efficacy against Caco-2 cells, with the flowering top extract being the most effective (about 90% activity at the highest concentration tested). In the brine shrimp lethality bioassay, the extracts exhibited no toxicity, indicating their potential safety.

Phenolic profile and biological properties of the leaves of Ficus vasta Forssk. (Moraceae) growing in Egypt.

BACKGROUND: Ficus vasta Forssk. (Moraceae) is traditionally used for the treatment of various ailments; nonetheless, this species has been poorly studied to date. This work aimed to characterize the phenolic profile and to evaluate the antioxidant and antimicrobial properties of a hydroalcoholic extract obtained from F. vasta leaves collected in Egypt. METHODS: The phenolic profile of the extract was characterized by HPLC-PDA/ESI-MS. The antioxidant properties were examined by different in vitro systems: DPPH test, reducing power and metal chelating activity assays. Moreover, the ability of the extract to protect Escherichia coli growth and survival from H2O2-induced oxidative stress was evaluated. The potential toxicity was investigated using Artemia salina lethality bioassay. Finally, the antimicrobial properties against a representative set of Gram-positive and Gram-negative bacterial strains and the yeast C. albicans were assayed by standard methods. RESULTS: By HPLC-PDA/ESI-MS analysis 12 compounds belonging to the groups of phenolic acids and flavonoids were identified. The extract exhibited strong radical scavenging activity in DPPH test (IC50 = 0.0672 ± 0.0038 mg/mL), reducing power (3.65 ± 0.48 ASE/mL) and chelating activity (IC50 = 0.801 ± 0.007 mg/mL). A total protection against H2O2-induced damage on E. coli was observed. No toxicity against A. salina was found (LC50 > 1000 µg/mL). The extract exhibited bacteriostatic activity against almost all the bacteria tested (MICs: 250-62.5 µg/mL). CONCLUSIONS: The obtained results demonstrate the potential of F. vasta leaves as safe sources of natural antioxidant and antimicrobial compounds.

Comparative study of the phenolic profile, antioxidant and antimicrobial activities of leaf extracts of five Juniperus L. (Cupressaceae) taxa growing in Turkey.

The purpose of this study was to conduct a comparative evaluation of some biological properties of methanol and water extracts of leaves of five Juniperus taxa growing in Turkey: J. communis L. var. communis (Jcc), J. communis L. var. saxatilis Pall. (Jcs), J. drupacea Labill. (Jd), J. oxycedrus L. subsp. oxycedrus (Joo), J. oxycedrus L. subsp. macrocarpa (Sibth. & Sm.) Ball. (Jom). The antioxidant properties were examined in vitro; both in the DPPH and in the reducing power tests, Joo methanol extract resulted the most active (IC50 = 0.09 ± 0.01mg/mL and ASE/mL = 2.56 ± 0.06). In the TBA assay, Jcs methanol extract exhibited the highest activity (IC50 = 4.39 ± 0.47 µg/mL). The extracts displayed bacteriostatic activity against Staphylococcus aureus, and Jd methanol extract resulted the most effective (MIC = 19.53 µg/mL); no effect on the S. aureus biofilm formation was observed. The extracts resulted non-toxic in the Artemia salina lethality bioassay. Finally, the phenolic profile of the methanol extracts was characterized by HPLC-PDA/ESI-MS.

Inula viscosa (L.) Aiton leaves and flower buds: Effect of extraction solvent/technique on their antioxidant ability, antimicrobial properties and phenolic profile.

This study was designed to establish the most effective solvent/technique for extracting antioxidant phytoconstituents from leaves and flower buds of Inula viscosa (L.) Aiton (Asteraceae) grown wild in Morocco. Maceration and hot extraction with methanol or water and Soxhlet ethanol extraction were utilized. The antioxidant potential was evaluated in vitro by DPPH, reducing power, and ferrous ions chelating activity assays. I. viscosa leaf and flower bud extracts displayed the strongest effect in the DPPH test, being the Soxhlet ethanol the most active ones (IC50 = 54.24 ± 0.21 µg/mL and 39.77 ± 0.23 µg/mL); thus, they were selected for further investigations. The antimicrobial efficacy of the Soxhlet ethanol extracts against ATCC and food isolates strains was assayed; the leaf extract showed the best activity, and Candida albicans was the most sensitive strain (MIC = 125 µg/mL). The extracts resulted non-toxic against Artemia salina. Among the phenolics characterised by HPLC-PDA-ESI-MS, hispidulin hexoside, patuletin and spinacetin were identified for the first time.

Estudo in vitro do potencial citotóxico da Annona muricata L / In vitro study of the cytotoxic potential of Annona muricata L

O presente trabalho tem como objetivo avaliar a atividade citotóxica do extrato etanólico da casca do caule (AMC) e folha (AMF) da Annona muricata Linn. Para a realização desse estudo, inicialmente foi verificada a atividade do extrato etanólico nas concentrações de 1000, 800, 600, 200 e 100 ?gmL-1 para AMF e concentrações de 200, 150, 100, 50, 10 ?gmL-1 para AMC através do ensaio de Artemia salina Leach que é considerado um bioensaio preliminar no estudo de extratos com forte atividade biológica e permite realizar a avaliação da toxicidade envolvendo apenas um parâmetro: vida ou morte. Posteriormente foi realizado o ensaio de citotoxicidade através do método do MTT (3-(4,5-dimetiltiazol-2yl)-2,5-difenil brometo de tetrazolina em linhagens de SF-295 (glioblastoma - humano), OVCAR-8 (ovário) HCT-116 (colón) e HL-60 (leucemia pormielocítica). Os extratos foram testados na concentração de 50 ?g/mL para o teste de citotoxicidade de concentração única para verificar ausência ou presença de atividade. Para a determinação da concentração inibitória (CI50), todas as amostras foram testadas em concentrações seriadas que variaram de 0,09 a 50 ?g/mL utilizando 2 como fator de diluição. No presente estudo, as duas amostras utilizadas através do ensaio de Artemia salina Leach apresentaram concentração letal (CL50) superiores a 80 ?gmL-1. A folha apresentou CL50 = 324, 07?gmL-1 e a casca do caule apresentou CL50 = 196, 04?gmL-1. Já através do teste do MTT os valores de concentração inibitória (CI50) variaram de 12,81 a 22,65 ?g/mL para AMF e de 0,09 a <5 ?g/mL para AMC, frente as diferentes linhagens tumorais avaliadas. Diante dos resultados obtidos para a casca e folhas de A. muricata, avaliadas neste trabalho através dos bioensaios de toxicidade com Artemia salina Leach e com as células tumorais SF- 295, OVCAR-8, HCT-116 e HL-60, pode-se verificar o potencial tóxico de ambas as amostras, destacando-se mais as cascas que tiveram um potencial citotóxico maior. Devido a este fato, novos experimentos devem ser conduzidos para investigar melhor o potencial antitumoral das cascas do caule de A. muricata .(AU)

This study aims to evaluate the cytotoxic activity of the ethanol extract of the stem bark (AMC) and leaf (MFA) of Annona muricata Linn. To conduct this study was initially verified the activity of the ethanol extract at concentrations of 1000, 800 , 600 , 200 and 100 ?gmL-1 for MFA and concentrations of 200 , 150 , 100 , 50 , 10 ?gmL-1 for AMC through assay of Artemia salina Leach which is considered a preliminary study on bioassay of extracts with strong biological activity. Subsequently, the cytotoxicity assay was performed using the MTT (method 3 - (4,5 -dimethylthiazol- 2YL ) -2,5- diphenyl bromide in tetrazolina lines SF- 295 (glioblastoma - human), OVCAR -8 (ovarian) HCT -116 (colon) and HL -60 (promyelocytic leukemia). The extracts were tested at a concentration of 50 ?g/mL for the single concentration cytotoxicity test to determine the presence or absence of activity. For determination of inhibitory concentration (IC50), all samples were tested in serial concentrations ranging from 0, 09 to 50 ?g/mL using 2 as the dilution factor. The present study, both samples used by testing Artemia salina Leach had higher LC50 80 ?gmL-1. Sheet presented LC50 = 324, 07 ?gmL-1 and stem bark showed LC50 = 196, 04 ?gmL-1. Already through MTT inhibitory concentration values (IC50) ranged from 12, 81 to 22,65 ?g/mL for MFA and from 0,09 to <5?g/ mL for AMC, evaluated against various tumor cell lines. Results obtained for the bark and leaves of A. muricata, evaluated in this work through toxicity bioassays with Artemia salina L and tumor cells SF-295, OVCAR-8, HCT-116 and HL-60, we can verify the toxic potential of both samples, especially more hulls which had a higher cytotoxic potential. Due to this fact, further experiments should be conducted to further investigate the antitumor potential of the stem bark of A. muricata.(AU)