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Plantain (Plantago L.) Species as Novel Sources of Flavonoid Antioxidants

To examine the antioxidant properties of methanol extracts of selected Plantago species (P. argentea Chaix., P. holosteum Scop., P. major L., P. maritima L., and P. media L.), various assays that measure free radical scavenging ability were carried out: DPPH, hydroxyl radical, superoxide anion, and nitric oxide scavenger capacity tests, reducing power (FRAP) assay, and Fe(2+)/ascorbate induced lipid peroxidation. In all of the tests extracts showed a potent antioxidant effect compared with BHT, a well-known synthetic antioxidant, and the extract of P. major, accepted as an official remedy. Besides, in examined extracts the total phenolic amount (ranging from 38.43 to 70.97 mg of GAE/g of dw) and the total flavonoid content (5.31-13.10 mg of QE/g of dw) were determined. Furthermore, the presence and content of selected flavonoids (luteolin-7-O-glucoside, apigenin-7-O-glucoside, luteolin, apigenin, rutin, and quercetin) were studied using LC-MS/MS technique. LC-MS/MS analysis showed noticeable qualitative and quantitative differences between the species according to which the examined Plantago species could be regarded as a possible new source of natural antioxidants. In this study three of the species examined, P. maritima, P. argentea, and P. holosteum, have been analyzed for the first time.

Quantitative determination of plant phenolics in Urtica dioica extracts by high-performance liquid chromatography coupled with tandem mass spectrometric detection

A method for quantification of 45 plant phenolics (including benzoic acids, cinnamic acids, flavonoid aglycones, C- and O-glycosides, coumarins, and lignans) in plant extracts was developed, based on reversed phase HPLC separation of extract components, followed by tandem mass spectrometric detection. The phenolic profile of 80% MeOH extracts of the stinging nettle (Urtica dioica L.) herb, root, stem, leaf and inflorescence was obtained by using this method. Twenty-one of the investigated compounds were present at levels above the reliable quantification limit, with 5-O-caffeoylquinic acid, rutin and isoquercitrin as the most abundant. The inflorescence extracts were by far the richest in phenolics, with the investigated compounds amounting 2.5-5.1% by weight. As opposed to this, the root extracts were poor in phenolics, with only several acids and derivatives being present in significant amounts. The results obtained by the developed method represent the most detailed U. dioica chemical profile so far.

Liquid chromatography/tandem mass spectrometry study of anti-inflammatory activity of plantain (Plantago L.) species.

To evaluate anti-inflammatory activity of selected Plantago species (P. lanceolata L. and P. major L.) an optimized in vitro test for determination of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibition potency was undertaken. By using intact cell system (platelets) as a source of COX-1 and 12-LOX enzymes and highly sensitive and specific LC-MS/MS technique for detection of main arachidonic acid metabolites formed by COX-1 and 12-LOX, this test provides efficient method for evaluation of anti-inflammatory potential of plant extracts and isolated compounds. Our results validated the well-known COX-1 inhibitory activity of P. lanceolata and P. major methanol extracts (concentration required for 50% inhibition (IC(50)) was 2.00 and 0.65 mg/ml, respectively). Furthermore, 12-LOX inhibitory activity of examined extracts was reported for the first time (IC(50)=0.75 and 1.73 mg/ml for P. lanceolata and P. major, respectively). Although renowned inhibitors, such as acetylsalicylic acid and quercetin showed higher activity, this study verifies P. lanceolata and P. major as considerable anti-inflammatory agents.

Comparative study of biological activities and phytochemical composition of two rose hips and their preserves: Rosa canina L. and Rosa arvensis Huds.

The aim of this study was to compare phenolic profile, vitamin C content, antioxidant, anti-inflammatory and cytotoxic activity of rose hips and the preserves (purée and jam) of two Rosa species: renowned Rosa canina L. and unexplored Rosa arvensis Huds. The liquid chromatography-tandem mass spectrometry analysis of 45 phenolics resulted in quantification of 14 compounds, with quercitrin, gallic and protocatechuic acids as the most dominant. High antioxidant potential of R. canina and a moderate activity of R. arvensis extracts were determined through several assays. Purée of both species and methanol extract of air-dried R. canina hips showed some anti-inflammatory (cyclooxygenase-1 and 12-lipooxygense inhibition potency) activity. Purée of R. canina exerted cytotoxic activity only against the HeLa cell line among several others (HeLa, MCF7, HT-29 and MRC-5). The presented results support traditional use of rose hips and their fruit preserves as food with health and nutritional benefits.

Phenolic profile, antioxidant, anti-inflammatory and cytotoxic activities of black (Tuber aestivum Vittad.) and white (Tuber magnatum Pico) truffles

The aim of this study was a comprehensive investigation on phenolic profile, antioxidant, anti-inflammatory and cytotoxic activity of the still insufficiently explored black summer truffles (Tuber aestivum Vittad.) and white (Tuber magnatum Pico) truffles. Methanol and water extracts, obtained by maceration and Soxhlet extraction, were examined. Forty-five phenolics was studied using LC-MS/MS: presence of 14 compounds was confirmed, with the most dominant being p-hydroxybenzoic acid, baicalein and kaempferol (T. aestivum), epicatechin and catechin (T. magnatum). Moderate antioxidant activity of both species was determined through several assays. Only T. magnatum showed anti-inflammatory potential by inhibiting COX-1 and 12-LOX pathway products synthesis. Methanol extracts exerted cytotoxicity against some tumour cell lines (HeLa, MCF7, HT-29), besides the prominent activity of water extracts towards breast adenocarcinoma (MCF7). To conclude, these results support further investigations of phytochemicals and biological activity towards verification of nutraceutical use of both species.

Binary and Tertiary Mixtures of Satureja hortensis and Origanum vulgare Essential Oils as Potent Antimicrobial Agents Against Helicobacter pylori

Essential oils possess strong antimicrobial activity, even against multiresistant Helicobacter pylori. Available therapies against H. pylori infection have multiple disadvantages, indicating a great need for a development of new therapeutics. The purpose of this study was to develop a potent natural product based anti‐H. pylori formulation. First, anti‐H. pylori activity of nine essential oils was determined, after which the most active oils were mixed in various ratios for further testing. Satureja hortensis, Origanum vulgare subsp. vulgare and O. vulgare subsp. hirtum essential oils expressed the highest activity (MIC = 2 μL mL−1). Their binary and ternary mixtures exhibited notably higher antimicrobial activity (MIC ≤ 2 μL mL−1). The most active was the mixture of S. hortensis and O. vulgare subsp. hirtum oils in volume ratio 2:1, which expressed 4 times higher activity than individual oils (MIC = 0.5 μL mL−1). According to GC‐MS, both oils in the mixture were characterized by high content of phenols (48–73%), with carvacrol as the main carrier of antimicrobial activity. Presented in vitro study pointed out binary mixture of S. hortensis and O. vulgare subsp. hirtum essential oils in volume ratio 2:1 as promising candidate for further in vivo studies targeting H. pylori infection. Copyright

Quantitative Determination of Aucubin in Seven Plantago Species Using HPLC, HPTLC, and LC-ESI-MS Methods

A reversed-phase high-performance liquid chromatographic (HPLC) method was developed and validated for the determination of aucubin in Plantago lanceolata. The analyses were carried out on Zorbax SB-C18 column with an aqueous phosphoric acid and acetonitrile gradient. The correlation coefficient of calibration curve showed good linearity (r > 0.9995), with average recoveries between 96.7 and 104.5%. The developed method was applied for quantification in P. atrata, P. bellardii, P. coronopus, P. holosteum, P. reniformis, and P. schwarzenbergiana. The aucubin content in plant extracts was compared by HPLC, HPTLC, and LC-ESI-MS techniques and no significant differences between the conducted methods were observed.

Phytochemical composition and in vitro functional properties of three wild rose hips and their traditional preserves

The aim of the present study was investigation of the phenolic profile, ascorbic acid content, antioxidant, anti-acetylcholinesterase, anti-inflammatory and cytotoxic activity of rose hips and the preserves (purée and jam) of three insufficiently examined Rosa species: Rosa dumalis Bechst., R. dumetorum Thuill. and R. sempervirens L. The liquid chromatography-tandem mass spectrometry analysis resulted in quantification of 14 of the 45 phenolic compounds examined, with ellagic acid as the most dominant. Notable antioxidant activity of all three species was confirmed through several assays. Moderate inhibition of acetylcholinesterase by extracts of all investigated Rosa species was observed. Several extracts of examined Rosa species demonstrated inhibition potency towards production of some monitored eicosanoids in cyclooxygenase-1 and 12-lipoxygenase pathways. Two R. sempervirens extracts exerted cytotoxic activity against HeLa and HT-29 cell lines, but were inactive towards MRC-5 and MCF7. The results support the potential of these rose hips as food with health-promoting properties.

The Effect of Plant Secondary Metabolites on Lipid Peroxidation and Eicosanoid Pathway

© 2012 Mimica-Dukic et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Effect of Plant Secondary Metabolites on Lipid Peroxidation and Eicosanoid Pathway

Polyphenolic profile, antioxidant and neuroprotective potency of grape juices and wines from Fruška Gora region (Serbia)

ABSTRACT This article reports novel and valuable data about three red (Cabernet Sauvignon, Frankovka and Merlot varieties), one rosé (Muscat Hamburg) and three white wines (Italian Riesling, Chardonnay and Sila), together with their related products (grape juices and young wines) produced in Fruška Gora vineyards, Serbia, concerning their in-depth polyphenolic profile and biological activities (antioxidant and neuroprotective). LC-MS/MS analysis of 47 phenolics showed that the phenolic acids, particularly ellagic (up to 8.99 mg/L, Cabernet Sauvignon) and caffeic acids (up to 2.83 mg/L, Frankovka) were generally more abundant in red wines. Also, the content of catechin and epicatechin (0.32–7.70 and 0.4–15.5 mg/L) as the leading flavonoids, malvidin 3-O-glucoside as the dominant anthocyanin (2.7–160 mg/L) or stilbene resveratrol (0.06–2.42 mg/L), were also higher in the red grape products. Overall antioxidant dominance of the red grape products, Merlot variety being the main one, was confirmed through several assays (DPPH• and •NO scavenging ability, reducing potential and inhibition of lipid peroxidation). In addition, grape juices and young wines, which are a good source of dietary natural antioxidants, were also evidenced. Furthermore, the optimised in vitro acetylcholinesterase inhibition assay was applied, indicating the grape juices and red wines as the most promising neuroprotective agents. Abbreviations: AAE: ascorbic acid equivalents; AChE: acetylcholinesterase; BHT: butylated hydroxytoluene; DPPH: 2,2-diphenyl-1-picrylhydrazyl; dw: dry weight; FA: fatty acids; FRAP: Ferric Reducing Antioxidant Power; LP: lipid peroxidation; MDA: malondialdehyde; SRM: selected reactions monitoring; PCA: principal component analysis; PG: propyl gallate; TBA: 2-thiobarbituric acid; TPTZ: 2,4,6-tripyridil-s-triazine.