Comparative metabolic profiling of olive leaf extracts from twelve different cultivars collected in both fruiting and flowering seasons.

Olea europaea is an economically significant crop native to Mediterranean countries. Its leaves exhibit several biological properties associated to their chemical composition. The aqueous ethanolic extracts of olive leaves from twelve different cultivars were analyzed by high performance liquid chromatography coupled to photodiode array and electrospray ionization mass spectrometry (HPLC/PDA/ESI-MS/MS). A total of 49 phytochemicals were identified in both positive and negative ionization modes. The identified compounds belonged to four classes of secondary metabolites including secoiridoids, flavonoids, pentacyclic triterpenoids and various phenolic compounds. Seasonal variation in chemical composition among the studied cultivars was apparent in autumn and spring. Secologanoside, oleuropein, hydroxy-oleuropein, demethyl oleuropein, gallocatechin, luteolin-O-hexoside, diosmetin, oleanolic acid and maslinic acid were detected in all cultivars in both seasons. Oleuropein-O-deoxyhexoside was tentatively identified for the first time in olive leaf extracts; detected only in the Spanish cultivar Picual (PIC) collected in spring. Also, dihydroxy-oxooleanenoic acid and hydroxy-oxooleanenoic acid, two bioactive pentacyclic triterpenes, were identified. Principle component analysis (PCA) showed good discrimination among the studied cultivars in terms of their botanical origin. This study is considered the first study for non-targeted metabolic profiling of different olive leaf cultivars cultivated in Egypt.

Dissection of Moringa oleifera leaf metabolome in context of its different extracts, origin and in relationship to its biological effects as analysed using molecular networking and chemometrics.

M. oleifera known as "miracle tree" is increasingly used in nutraceuticals for the reported health effects and nutritional value of its leaves. This study presents the first metabolome profiling of M. oleifera leaves of African origin using different solvent polarities via HR-UPLC/MS based molecular networking followed by multivariate data analyses for samples classification. 119 Chemicals were characterized in both positive and negative modes belonging to 8 classes viz. phenolic acids, flavonoids, peptides, fatty acids/amides, sulfolipids, glucosinolates and carotenoids. New metabolites i.e., polyphenolics, fatty acids, in addition to a new class of sulfolipids were annotated for the first time in Moringa leaves. In vitro anti-inflammatory and anti-aging bioassays of the leaf extracts were assessed and in correlation to their metabolite profile via multivariate data analyses. Kaempferol, quercetin and apigenin-O/C-glycosides, fatty acyl amides and carotenoids appeared crucial for biological activities and leaves origin discrimination.

Diverse host-associated fungal systems as a dynamic source of novel bioactive anthraquinones in drug discovery: Current status and future perspectives.

BACKGROUND: Despite, a large number of bioactive anthraquinones (AQs) isolated from host-living fungi, only plant-derived AQs were introduced in the global consumer markets. Host-living fungi represents renewable and extendible resources of diversified metabolites to be exploited for bioactives production. Unique classes of AQs from fungi include halogenated and steroidal AQs, and absent from planta are of potential to explore for biological activity against urging diseases such as cancer and multidrug-resistant pathogens. The structural diversity of fungal AQs, monomers, dimers, trimers, halogenated, etc… results in a vast range of pharmacological activities. AIM OF REVIEW: The current study capitalizes on uncovering the diversity and distribution of host-living fungal systems producing AQs in different terrestrial ecosystems ranging from plant endophytes, lichens, animals and insects. Furthermore, the potential bioactivities of fungal derived AQs i.e., antibacterial, antifungal, antiviral (anti-HIV), anticancer, antioxidant, diuretic and laxative activities are assembled in relation to their structure activity relationship (SAR). Analyzing for structure-activity relationship among fungal AQs may facilitate bioengineering of more potential analogues. Withal, elucidation of AQs biosynthetic pathways in fungi is discussed from different fungal hosts to open up new possibilities for potential biotechnological applications. Such comprehensive review unravels terrestrial host-living fungal systems as a treasure trove in drug discovery, in addition to future perspectives and trends for their exploitation in pharmaceutical industries. KEY SCIENTIFIC CONCEPTS OF REVIEW: Such comprehensive review unravels terrestrialhost-living fungal systems as a treasure trove in drug discovery, in addition to future perspectives and trends for their exploitation in pharmaceutical industries.

Nanoencapsulated Extract of a Red Seaweed (Rhodophyta) Species as a Promising Source of Natural Antioxidants.

Marine seaweed species represent an important source of bioactive compounds possessing antioxidant activity. This study aimed at evaluating the antioxidant capacity of the Jania rubens algal extract by means of two antioxidant assays, i.e., 2,2-diphenyl-1-picrylhydrazyl and ferric-reducing antioxidant power. The seaweeds' total phenolic and flavonoid contents were also assayed as markers of antioxidant activity. To identify active agents responsible for the antioxidant activity, gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry were used for comprehensive metabolites characterization. To enhance the Jania rubens efficacy, the extract was nanoencapsulated using an ionic gelation method by means of high-pressure homogenization. The optimum nanoformulation had a particle size of 161 nm, a ζ potential of 31.2 mV, a polydispersity index of 0.211, and entrapment efficiency of 99.7%. The in vitro phytochemicals' release profiles of Jania rubens chitosan nanoparticles in comparison to the concentration of the raw algal extract were studied by the dialysis bag diffusion method revealing that the extract was released in a controlled pattern. The results indicated the potential advantages of the encapsulated Jania rubens extract, with its potent antioxidant activity, for use in different applications where sustained release is useful.

Metabolic and biotransformation effects on dietary glucosinolates, their bioavailability, catabolism and biological effects in different organisms.

Glucosinolate-producing plants have long been recognized for both their distinctive benefits to human nutrition and their resistance traits against pathogens and herbivores. Despite the accumulation of glucosinolates (GLS) in plants is associated with their resistance to various biotic and abiotic stresses, the defensive and biological activities of GLS are commonly conveyed by their metabolic products. In view of this, metabolism is considered the driving factor upon the interactions of GLS-producing plants with other organisms, also influenced by plant and plant attacking or digesting organism characteristics. Several microbial pathogens and insects have evolved the capacity to detoxify GLS-hydrolysis products or inhibit their formation via different means, highlighting the relevance of their metabolic abilities for the plants' defense system activation and target organism detoxification. Strikingly, some bacteria, fungi and insects can likewise produce their own myrosinase (MYR)-like enzymes in one of the most important adaptation strategies against the GLS-MYR plant defense system. Knowledge of GLS metabolic pathways in herbivores and pathogens can impact plant protection efforts and may be harnessed upon for genetically modified plants that are more resistant to predators. In humans, the interest in the implementation of GLS in diets for the prevention of chronic diseases has grown substantially. However, the efficiency of such approaches is dependent on GLS bioavailability and metabolism, which largely involves the human gut microbiome. Among GLS-hydrolytic products, isothiocyanates (ITC) have shown exceptional properties as chemical plant defense agents against herbivores and pathogens, along with their health-promoting benefits in humans, at least if consumed in reasonable amounts. Deciphering GLS metabolic pathways provides critical information for catalyzing all types of GLS towards the generation of ITCs as the biologically most active metabolites. This review provides an overview on contrasting metabolic pathways in plants, bacteria, fungi, insects and humans towards GLS activation or detoxification. Further, suggestions for the preparation of GLS containing plants with improved health benefits are presented.

Metabolites profiling reveals gut microbiome-mediated biotransformation of green tea polyphenols in the presence of N-nitrosamine as pro-oxidant.

The gut microbiome contributes to host physiology and nutrition metabolism. The interaction between nutrition components and the gut microbiota results in thousands of metabolites that can contribute to various health and disease outcomes. In parallel, the interactions between foods and their toxicants have captured increasing interest due to their impact on human health.  Taken together, investigating dietary interactions with endogenous and exogenous factors and detecting interaction biomarkers in a specific and sensitive manner is an important task. The present study sought  to identify for the first time the metabolites produced during the interaction of diet-derived toxicants e.g., N-nitrosamines with green tea polyphenols, using liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). In addition, the metabolic products resulting from the incubation of green tea with a complex gut microbiome in the presence of N-nitrosamine were assessed in the same manner. The quinone products of (epi)catechin, quercetin, and kaempferol were identified when green tea was incubated with N-nitrosamine only; whereas, incubation of green tea with N-nitrosamine and a complex gut microbiome prevented the formation of these metabolites. This study provides a new perspective on the role of gut microbiome in protecting against potential negative interactions between food-derived toxicants and dietary polyphenols.

Quality assessment of leaf extracts of 12 olive cultivars and impact of seasonal variation based on UV spectroscopy and phytochemcial content using multivariate analyses.

INTRODUCTION: Recently, focus has been made on the health-oriented uses of olive leaves, a byproduct of olive production, as a potential source of antioxidants. Oleuropein is one of the phenolic components in olive leaves known for its high antioxidant value. OBJECTIVE: The main aim of the current study was constructing a model for the quality assessment of olive leaves and their potential phytochemical content and hence biological value as well. The phytochemical variation in olive leaves in both flowering (spring) and fruiting seasons (autumn) was also investigated. METHODS: In this study, the leaves of 12 different olive cultivars from different geographical origins growing in Egypt were assessed for their oleuropein content, total flavonoid (TF) content and total polyphenol (Pph) content in spring and autumn via ultraviolet (UV) spectroscopy and high-performance liquid chromatography (HPLC) coupled to multivariate data analyses. The antioxidant activity of olive leaf extracts was assessed using 2,2'-diphenyl-1-picrylhydrazyl (DPPH) assay. RESULTS: Higher levels of oleuropein, TF and Pph content were found in spring with the highest oleuropein content in the Spanish cultivar; Manzanillo, followed by the Italian cultivar Coratina and the Egyptian Agizi Okasi (218.94, 151.58 and 122.18 mg/100 g of dried leaf extract, respectively). UV spectra was also measured and the collected data were coupled to multivariate analyses showing clustering of cultivars with common geographical origin. CONCLUSION: Our findings emphasised the influence of collection time and type of cultivar on the chemical profile of olive leaves. The model presented herein, serves for the quality assessment of olive leaves based on their phytochemical profile.

Phytochemical and Biological Evaluation of a Newly Designed Nutraceutical Self-Nanoemulsifying Self-Nanosuspension for Protection and Treatment of Cisplatin Induced Testicular Toxicity in Male Rats.

The incorporation of cisplatin (CP) as a cytotoxic antineoplastic agent in most chemotherapeutic protocols is a challenge due to its toxic effect on testicular tissues. Natural compounds present a promising trend in research, so a new nutraceutical formulation (NCF) was designed to diminish CP spermatotoxicity. A combination of three nutraceutical materials, 250 mg Spirulina platensis powder (SP), 25 mg Tribulus terrestris L. extract (TT), and 100 mg fish oil (FO) were formulated in self-nanoemulsifying self-nanosuspension (SNESNS). SP was loaded into the optimized self-nanoemulsifying system (30% FO, 50% span 80/cremophor EL and 20% isopropanol) and mixed with TT aqueous solution to form SNESNS. For the SP, phytochemical profiling revealed the presence of valuable amounts of fatty acids (FAs), amino acids, flavonoids, polyphenols, vitamins, and minerals. Transmission electron microscopy (TEM) and particle size analysis confirmed the formation of nanoemulsion-based nanosuspension upon dilution. Method validation of the phytochemical constituents in NCF has been developed. Furthermore, NCF was biologically evaluated on male Wistar rats and revealed the improvement of spermatozoa, histopathological features, and biochemical markers over the CP and each ingredient group. Our findings suggest the potential of NCF with SNESNS as a delivery system against CP-induced testicular toxicity in male rats.

Novel trends and applications of multidimensional chromatography in the analysis of food, cosmetics and medicine bearing essential oils.

The analysis of aroma composition in scented plants and natural products to probe their quality and safety is an important topic, particularly when such ingredients are intended for long term use as in food or cosmetic products. The development of fast, comprehensive, and effective analytical tools for essential oil analysis in such complex mixtures is of interest and moreover when present in blend of several oils, as typical in case of nutraceuticals or cosmetics. Comprehensive studies of volatiles could be achieved via coupling the separation power of multidimensional chromatography with selective detectors as mass spectrometers. This strategy enables high-throughput and global analysis of hundreds of metabolites in a single step. Different multidimensional setups such as GC × GC and LC-GC in addition to various chemometric approaches applied for essential oils analysis form a fundamental part of this review. Asides, applications of multidimensional chromatography for essential oils chemotyping, enantio-separation, quality control and adulteration detection in the different matrices are also presented.

Unraveling the metabolome composition and its implication for Salvadora persica L. use as dental brush via a multiplex approach of NMR and LC-MS metabolomics.

Salvadora persica L. (toothbrush tree, Miswak) is well recognized in most Middle Eastern and African countries for its potential role in dental care, albeit the underlying mechanism for its effectiveness is still not fully understood. A comparative MS and NMR metabolomics approach was employed to investigate the major primary and secondary metabolites composition of S. persica in context of its organ type viz., root or stem to rationalize for its use as a tooth brush. NMR metabolomics revealed its enrichment in nitrogenous compounds including proline-betaines i.e., 4-hydroxy-stachydrine and stachydrine reported for the first time in S. persica. LC/MS metabolomics identified flavonoids (8), benzylurea derivatives (5), butanediamides (3), phenolic acids (8) and 5 sulfur compounds, with 21 constituents reported for the first time in S. persica. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) of either NMR or LC/MS dataset clearly separated stem from root specimens based on nitrogenous compounds abundance in roots and is justifying for its preference as toothbrush versus stems. The presence of betaines at high levels in S. persica (9-12 µg/mg dry weight) offers novel insights into its functioning as an osmoprotectant that maintains the hydration of oral mucosa. Additionally, the previously described anti-inflammatory activity of stachydrine along with the antimicrobial effects of sulfonated flavonoids, benzylisothiocynate and ellagic acid derivatives are likely contributors to S. persica oral hygiene health benefits. Among root samples, variation in sugars and organic acids levels were the main discriminatory criterion. This study provides the first standardization of S. persica extract using qNMR for further inclusion in nutraceuticals.

How do biocatalysis and biotransformation affect Citrus dietary flavonoids chemistry and bioactivity? A review.

Flavonoids, especially flavanones, flavones and polymethoxyflavones (PMFs) are distinctive bioactive compounds of Citrus fruits. Citrus peel and juice by-products potentially represent rich sources of these Citrus flavonoids that exhibit a myriad of biological activities both in in vitro and in vivo systems. Recently, much attention has been made toward biotransformation processes as a promising tool for the structural modification of natural products to be used in the drug and food industries along with its role in solving pollution problems related to Citrus by-products disposal. In this article, we present a state of the art review on both in vivo and in vitro biotransformation processes of Citrus juice and waste carried out by microorganisms, plant cell cultures, animal and human liver microsomes targeting its flavonoids composition. Such review highlights the main metabolic pathways for enzymatic and microbial reactions involved in these processes and suggest for reactions that need to be more capitalized for a wider application in industrial bioprocesses of Citrus by-products. Biotransformation and biocatalysis applications included employment of citrus by-products enriched in flavonoids as a low-cost, economical and natural sources of sugar substitutes, antifungal, anticancer drugs, hydrolyzable enzymes, probiotics and flavonoid aglycones. Further, biological effects of the biotransformed metabolites are discussed in relation to its parent compound highlighting potentials and or any limitations for each reaction type. Applications covered in Citrus biotransformation include for nutraceutical/food and cosmetics industries.

Metabolomics driven analysis of 11 Portulaca leaf taxa as analysed via UPLC-ESI-MS/MS and chemometrics.

Portulaca oleracea, commonly known as purslane, is a popular plant of considerable value for its nutritive composition as well as traditional medicinal uses. P. oleracea is reported to possess neuroprotective, antimicrobial, antidiabetic, antioxidant, anti-inflammatory, antiulcerogenic, and anticancer activities. Three taxa of P. oleracea L. (P. oleracea, P. rausii and P. granulatostellulata) are grown as mixed populations in several locations in Egypt. The close morphological similarities among these taxa warrants development of methods for their correct identification or classification. We aimed in this study to assess metabolome differences among three P. oleracea taxa via ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) in the context of their genetic diversity and/or geographical origin. A total of 85 metabolites were identified including 6 amino acids, 22 phenolic compounds, 16 alkaloids, and 11 fatty acids characterized based on their MSn and UV spectra. Methoxylated flavone glycosides, O-flavonoids, C-flavonoids and four previously undescribed cyclodopa alkaloids are reported in P. oleracea for the first time. Multivariate data analyses were used for samples classification and revealing that cyclodopa alkaloids (oleracein A, C, K and N) contributed the most for accessions classification. To the best of our knowledge, this study presents the first metabolite profile of Portulaca and its compositional differences that provide chemical based evidence for its nutritive and/or health benefits.