From sample preparation to NMR‐based metabolic profiling in food commodities: The case of table olives

Nuclear magnetic resonance (NMR)‐based metabolic profiling has been widely used in food and plant sciences. Despite its simplicity and inherent reproducibility, the determination of the appropriate pre‐processing procedures greatly affects the obtained metabolic profile. The current study represents a detailed guide of use for untargeted NMR‐based metabolic profiling of table olives (Olea europaea L.). Greek Kalamon table olives from different geographical origins were selected as reference materials. Differently treated samples were extracted using different solvents and/or solvent systems. Chemical profiles were evaluated with high‐performance thin layer chromatography (HPTLC). Different deuterated solvents and sample concentrations were evaluated for the recording of optimal quality spectra. The methanol extract of freeze‐dried table olives was found to contain the most representative secondary metabolites, in higher concentrations, as well. The optimal deuterated solvent for the NMR analysis was methanol‐d4, while final sample concentration should be within the range of 10 to 15 mg/mL. Multivariate data analysis was also used to estimate and confirm the variation and clustering caused by different characteristics of the samples. Results of the present study make evident the necessity for thorough planning and method development prior to any extensive metabolomic study based on NMR spectroscopy. Pre‐processing and sample preparation stages seemed to greatly affect the metabolic profile and spectral quality in the case of table olives, which by extrapolation could apply to other food commodities. Nevertheless, the nature of the samples must be fully described in general, in order to proceed to solid conclusions

Differential effect of Pistacia vera extracts on experimental atherosclerosis in the rabbit animal model: an experimental study

<p>Abstract</p> <p>Background</p> <p>Lipid-enriched diets and oxidative stress are risk factors for the development of atherosclerosis. The effects of the methanolic (ME) and cyclohexane (CHE) extracts of the <it>Pistacia vera </it>nut, often included in the Mediterranean diet, were studied in the rabbit model of atherosclerosis.</p> <p>Methods and results</p> <p>Twenty-four New Zealand White rabbits received atherogenic diet (Control Group), supplemented with ME (Group ME) or CHE (Group CHE) for 3 months. Previously, a GC-MS and a UHPLC LC-DAD-ESI(-)-HRMS/MS method were developed to investigate the extracts' chemical profiles. Blood samples at baseline and monthly determined lipid profile, lipid peroxidation and liver function. The aorta, myocardium and liver were examined histologically at 3 months.</p> <p>Groups ME and CHE had significantly higher HDL- and non-significantly lower LDL-cholesterol median % changes from baseline than the Control Group. Triacylglycerol was significantly higher in Group CHE vs. Control. MDA values were significantly lower in Group ME vs. Control and CHE. ALT and AST were significantly higher in Group CHE vs. Control. γ-GT was lower in Group ME vs. Control. Aortic intimal thickness was significantly less in Groups ME and CHE vs. Control; Group ME atherosclerotic lesions were significantly less extensive vs. Groups Control and CHE. Only Group CHE had significant liver fatty infiltration.</p> <p>Conclusions</p> <p>During short-term administration concomitantly with atherogenic diet, both <it>P. vera </it>extracts were beneficial on HDL-, LDL-cholesterol and aortic intimal thickness. The ME additionally presented an antioxidant effect and significant decrease of aortic surface lesions. These results indicate that <it>P. vera </it>dietary inclusion, in particular its ME, is potentially beneficial in atherosclerosis management.</p

Phytochemical analysis and antioxidant potential of the phytonutrient-rich decoction of Cichorium spinosum and C. intybus.

The Cretan diet as the basis of the Mediterranean diet, has provided traditional remedies for the general well-being through the long-established consumption of cooked wild greens and vegetables. The intake of water decoctions of Cichorium spinosum and Cichorium intybus in the context of the daily dietary regime in Greece, has been long associated with “liver detoxifying” properties. In the current study, we performed an in depth investigation of the water decoctions traditionally prepared from C. spinosum and C. intybus, through qualitative UHPLC-HRMS profiling and direct quantification of cichoric and caftaric acid, as major antioxidant components of the decoction. In addition, we developed a one-step Countercurrent Chromatography method for the isolation of the two phenolic acids, along with a sulfoconjugate sesquiterpene lactone present only in the Cretan C. spinosum. All water decoctions were found not cytotoxic in human fibroblasts, whereas they all significantly reduce the intracellular reactive oxygen species, in consistency with the major presence of strong antioxidant compounds such as cichoric acid. This work demonstrates that the intake of decoction in doses suggested by the Greek traditional use is comparable to the ingestion of a phytomedical preparation of antioxidants. These results contribute to our current knowledge on the beneficial health effect of the Cretan diet

Traditional uses, phytochemistry and pharmacology of Chios mastic gum (Pistacia lentiscus var. Chia, Anacardiaceae): A review

Ethnopharmacological relevance Chios mastic gum constitutes a unique Greek product, produced exclusively in the southern part of the island of Chios. References about its use from local populations for the treatment of gastrointestinal disorders or as a cosmetic agent can even be encountered in ancient texts of Galen, Theophrastus and Dioscorides. Nowadays, this versatile resin has been rediscovered, not only as a traditional remedy and aromatic agent, but as a potent phytotherapeutic product with various biological properties. Aim of the study: The aim of this study is to quote the summation of the ethnopharmacology, phytochemical profile and pharmacological properties of the resin of Pistacia lentiscus var. Chia and thus provide the scientific community with a summary of the research conducted so far. Furthermore, perspectives and uses are being discussed and studied so as to broaden the field of its applications. Materials and methods A comprehensive review of the literature on Pistacia lentiscus var. Chia was performed using as resources scientific databases such as Scopus, Sciencedirect, Pubmed and Web of science, studies and traditional books provided by the Chios Mastiha Growers Association as well as PhD and Master’ s theses. Results Chios mastic gum has been used as a traditional medicine over the last 2500 years. More than 120 chemical compounds have been identified in the resin and the major components are a natural polymer, acidic and neutral triterpenes and volatile secondary metabolites. Several plant extracts and compounds have been studied for their antibacterial, anti-inflammatory, antioxidant, anti-ulcer, anti-diabetic, cardioprotective and anti-cancer properties in vitro and in vivo. Clinical interventions and trials have also showed the therapeutic potential of Chios mastic gum. In 2015 Pistacia lentiscus L., resin (mastic) was recognized as a herbal medicinal product with traditional use by the European Medicines Agency (EMA) with two therapeutic indications (mild dyspeptic disorders &amp; skin inflammation/healing of minor wounds). Over the last years, Chios mastic gum is widely involved in medicinal products, food supplements and cosmetics and has become object of study, also in the field of Pharmacotechnology. Conclusions Chios mastic&apos;s beneficial properties have been demonstrated in the treatment of gastrointestinal disorders, wound healing, skin inflammations, plasma lipid and blood sugar reduction and oral care. These properties are attributed to triterpenes and volatile compounds. However, because of the resin&apos;s chemical complexity and the lack of commercial standards for its main compounds, there is a notable gap in literature concerning the biological evaluation of CMG&apos;s isolated components. Therefore, future research should focus on the development of efficient extraction, isolation and analysis techniques in order to unravel CMG&apos;s full pharmacological potential

Thermal, dynamic and structural properties of drug AT1 antagonist olmesartan in lipid bilayers

It is proposed that AT1 antagonists (ARBs) exert their biological action by inserting into the lipid membrane and then diffuse to the active site of AT1 receptor. Thus, lipid bilayers are expected to be actively involved and play a critical role in drug action. For this reason, the thermal, dynamic and structural effects of olmesartan alone and together with cholesterol were studied using differential scanning calorimetry (DSC), 13C magicangle spinning (MAS) nuclear magnetic resonance (NMR), cross-polarization (CP) MAS NMR, and Raman spectroscopy as well as small- and wide angle X-ray scattering (SAXS and WAXS) on dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles. 13C CP/MAS spectra provided direct evidence for the incorporation of olmesartan and cholesterol in lipid bilayers. Raman and X-ray data revealed how both molecules modify the bilayer&apos;s properties. Olmesartan locates itself at the head-group region and upper segment of the lipid bilayers as 13C CP/MAS spectra show that its presence causes significant chemical shift changes mainly in the A ring of the steroidal part of cholesterol. The influence of olmesartan on DPPC/cholesterol bilayers is less pronounced. Although, olmesartan and cholesterol are residing at the same region of the lipid bilayers, due to their different sizes, display distinct impacts on the bilayer&apos;s properties. Cholesterol broadens significantly the main transition, abolishes the pre-transition, and decreases the membrane fluidity above the main transition. Olmesartan is the only so far studied ARB that increases the gauche:trans ratio in the liquid crystalline phase. These significant differences of olmesartan may in part explain its distinct pharmacological profile

EXPLORATION OF DIETARY PLANTS FROM WORLDWIDE BIODIVERSITY AIMING THE DISCOVERY OF ACTIVE AGENTS WITH ANTI-AGEING EFFECTS

Dietary interventions obtain a profound impact on human lifespan, while constitute edible plants a promising source of chemical entities that promotes healthy ageing1. Maqui (Aristotelia chilensis, Elaeocarpaceae [Molina] Stunz), is a small tree native to South Chile, deeply rooted in traditional dietary culture of indigenous. Moreover, carob tree (Ceratonia siliqua, Leguminosae L.) has been grown since antiquity in most countries of Mediterranean region, composing a staple in the diet of people in times of famine. Maqui berries and carob pods, have drawn attention due to their potential health benefits, which are largely attributed to their rich polyphenolic content. A high-throughput phytochemical investigation of methanol and water extracts of the edible parts, berries and pods respectively, was performed. Fingerprinting procedures using HPTLC showed that both plants had rich chemical profile, while phenolic compounds prevailed. Profiling and dereplication procedures employing hyphenated techniques (HPLC/UHPLC-PDA, LC-(ESI)-HRMS/MS) were also applied and various putative bioactive compounds were identified. Hence, analytical and preparative techniques afforded compounds belonging to anthocyanins, flavonoids, flavonoid glucosides and phenolic acids, followed by structure elucidation (1D &amp; 2D NMR). The in vivo evaluation of anti-oxidant and anti-ageing properties of maqui and carob pods water extracts in Drosophila melanogaster flies revealed, that oral administration of both activate the Nrf2/Keap1 antioxidant pathway and proteasome peptidase activities. Complementary to the above, the metabolism of both dietary fruit extracts was investigated in vitro using rat liver microsomes. Conclusively, maqui and carob pod water extracts could be a potent source of anti-oxidant/ anti-ageing nutraceuticals

Olive oil with high polyphenolic content induces both beneficial and harmful alterations on rat redox status depending on the tissue

Olive oil (OO) possesses a predominant role in the diet of Mediterranean countries. According to a health claim approved by the European Food Safety Authority, OO protects against oxidative stress‑induced lipid peroxidation in human blood, when it contains at least 5 mg of hydroxytyrosol and its derivatives per 20 g. However, studies regarding the effects of a total OO biophenols on redox status in vivo are scarce and either observational and do not provide a holistic picture of their action in tissues. Following a series of in vitro screening tests an OO containing biophenols at 800 mg/kg of OO was administered for 14 days to male Wistar rats at a dose corresponding to 20 g OO/per day to humans. Our results showed that OO reinforced the antioxidant profile of blood, brain, muscle and small intestine, it induced oxidative stress in spleen, pancreas, liver and heart, whereas no distinct effects were observed in lung, colon and kidney. The seemingly negative effects of OO follow the recently formulated idea in toxicology, namely the real life exposure scenario. This study reports that OO, although considered a nutritional source rich in antioxidants, it exerts a tissues specific action when administered in vivo

Extracts from the Mediterranean Food Plants Carthamus lanatus, Cichorium intybus, and Cichorium spinosum Enhanced GSH Levels and Increased Nrf2 Expression in Human Endothelial Cells

The Mediterranean diet is considered to prevent several diseases. In the present study, the antioxidant properties of six extracts from Mediterranean plant foods were assessed. The extracts’ chemical composition analysis showed that the total polyphenolic content ranged from 56 to 408 GAE mg/g dw of extract. The major polyphenols identified in the extracts were quercetin,luteolin, caftaric acid, caffeoylquinic acid isomers, and cichoric acid. The extracts showed in vitro high scavenging potencyagainst ABTS•+and O2•−radicals and reducing power activity. Also, the extracts inhibited peroxyl radical-induced cleavage ofDNA plasmids. The three most potent extracts, Cichorium intybus, Carthamus lanatus, and Cichorium spinosum, inhibited OH•-induced mutations in Salmonella typhimurium TA102 cells. Moreover, C. intybus ,C. lanatus, and C. spinosum extracts increased the antioxidant molecule glutathione (GSH) by 33.4, 21.5, and 10.5% at 50μg/ml, respectively, in human endothelialEA.hy926 cells.C. intybusextract was also shown to induce in endothelial cells the transcriptional expression of Nrf2 (the majortranscription factor of antioxidant genes), as well as of antioxidant genes GCLC, GSR, NQO1, and HMOX1. In conclusion, theresults suggested that extracts from edible plants may prevent diseases associated especially with endothelium damag