A multiplex metabolomic approach for quality control of Spirulina supplement and its allied microalgae (Amphora & Chlorella) assisted by chemometrics and molecular networking.

Microalgae species are of economic importance regarded as "green gold" being rich in bioactive compounds. Spirulina and Chlorella are the most popular microalgal species and are marketed as healthy food supplements. At the same time, Amphora holds potential as a source of healthy lipids and essential fatty acids. Yet, there are considerable variations in their reported chemical composition, and less is known about their compositional differences. A multiplexed metabolomic approach was adopted for the quality control (QC) of Spirulina supplements and to compare its constitutive metabolome to Chlorella and Amphora. The adopted protocol comprised gas chromatography-mass spectrometry (GC-MS), ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS), and ultraviolet-visible spectrophotometry (UV/Vis) for mapping their primary and secondary metabolome. Interestingly, UPLC-HRMS/MS analysis delineated the abundance of fatty acids in Amphora versus glycolipids enrichment in Spirulina, and porphyrins were the main pigments identified in Spirulina, with scarce occurrence in Chlorella. Orthogonal projections to latent structures discriminant analysis (OPLS-DA) analysis of GC-MS data set revealed palmitic acid, 3-mannobiose, and glyceryl-glycoside as being most enriched in Spirulina, versus sucrose and leucine in Chlorella and Amphora, respectively. Despite being of low discriminatory potential, UV/Vis OPLS-DA modeling showed that Spirulina was distinguished with the UV absorbances of carotenoids and chlorophyll pigments, as indicated by its OPLS-DA derived S-plot. Our study provides a QC approach for the analysis of the microalgal species and poses alternative spectral and compositional markers for their discrimination.

Bioactives and functional food ingredients with promising potential for the management of cerebral and myocardial ischemia: a comprehensive mechanistic review.

Ischemia is a deadly disease featured by restricted perfusion to different organs in the body. An increase in the accumulation of reactive oxygen species and cell debris is the driving force for inducing many oxidative, inflammatory and apoptotic signaling pathways. However, the number of therapeutics existing for ischemic stroke patients is limited and there is insufficient data on their efficiency, which warrants the search for novel therapeutic candidates from natural sources. Herein, a comprehensive survey was done on the reported functional food bioactives (ca. 152 compounds) to manage or protect against health consequences of myocardial and cerebral ischemia. Furthermore, we reviewed the reported mechanistic studies for their anti-ischemic potential. Subsequently, network pharmacology- and in silico-based studies were conducted using the reported myocardial and cerebral ischemia-relevant molecular targets to study their complex interactions and highlight key targets in disease pathogenesis. Subsequently, the most prominent 20 compounds in the literature were used in a comprehensive in silico-based analysis (inverse docking, ΔG calculation and molecular dynamics simulation) to determine other potential targets for these compounds and their probable interactions with different signaling pathways relevant to this disease. Many functional food bioactives, belonging to different chemical classes, i.e., flavonoids, saponins, phenolics, alkaloids, iridoids and carotenoids, were proven to exhibit multifactorial effects in targeting the complex pathophysiology of ischemic conditions. These merits make them valuable therapeutic agents that can outperform the conventional drugs, and hence they can be utilized as add-ons to the conventional therapy for the management of different ischemic conditions; however, their rigorous clinical assessment is necessary.

Metabolomics-based profiling of 4 avocado varieties using HPLC-MS/MS and GC/MS and evaluation of their antidiabetic activity.

Seven avocado "Persea americana" seeds belonging to 4 varieties, collected from different localities across the world, were profiled using HPLC-MS/MS and GC/MS to explore the metabolic makeup variabilities and antidiabetic potential. For the first time, 51 metabolites were tentatively-identified via HPLC-MS/MS, belonging to different classes including flavonoids, biflavonoids, naphthodianthrones, dihydrochalcones, phloroglucinols and phenolic acids while 68 un-saponified and 26 saponified compounds were identified by GC/MS analysis. The primary metabolic variabilities existing among the different varieties were revealed via GC/MS-based metabolomics assisted by unsupervised pattern recognition methods. Fatty acid accumulations were proved as competent, and varietal-discriminatory metabolites. The antidiabetic potential of the different samples was explored using in-vitro amylase and glucosidase inhibition assays, which pointed out to Gwen (KG) as the most potent antidiabetic sample. This could be attributed to its enriched content of poly-unsaturated fatty acids and polyphenolics. Molecular docking was then performed to predict the most promising phytoligands in KG variety to be posed as antidiabetic drug leads. The highest in-silico α-amylase inhibition was observed with chrysoeriol-4'-O-pentoside-7-O-rutinoside, apigenin-7-glucuronide and neoeriocitrin which might serve as potential drug leads for the discovery of new antidiabetic remedies.

Authentication of saffron spice accessions from its common substitutes via a multiplex approach of UV/VIS fingerprints and UPLC/MS using molecular networking and chemometrics.

Saffron is a spice revered for its unique flavor and health attributes often subjected to fraudulence. In this study, molecular networking as a visualization tool for UPLC/MS dataset of saffron and its common substitutes i.e. safflower and calendula (n = 21) was employed for determining genuineness of saffron and detecting its common substitutes i.e. safflower and calendula. Saffron was abundant in flavonol-O-glycosides and crocetin esters versus richness of flavanones/chalcones glycosides in safflower and cinnamates/terpenes in calendula. OPLS-DA identified differences in UPLC/MS profiles of different saffron accessions where oxo-hydroxy-undecenoic acid-O-hexoside was posed as saffron authentication marker and aided in discrimination between Spanish saffron of high quality from its inferior grade i.e. Iranian saffron along with crocetin di-O-gentiobiosyl ester and kaempferol-O-sophoroside. Kaempferol-O-neohesperidoside and N,N,N,-p-coumaroyl spermidine were characteristic safflower metabolites, whereas, calendulaglycoside C and di-O-caffeoyl quinic acid were unique to calendula. UV/VIS fingerprint spectral regions of picrocrocin (230-260 nm) and crocin derivatives (400-470 nm) were posed as being discriminatory of saffron authenticity and suggestive it can replace UPLC/MS in saffrom quality determination.

In silico study of potential anti-SARS cell entry phytoligands from Phlomis aurea: a promising avenue for prophylaxis.

Aim: The severity of COVID-19 has raised a great public health concern evoking an urgency for developing multitargeted therapeutics. Phlomis species was ethno-pharmacologically practiced for respiratory ailments. Materials & methods: An array of 15 phytoligands previously isolated from Phlomis aurea were subjected to molecular docking to explore their potential SARS-CoV-Spike-angiotensin-converting enzyme 2 complex inhibition, that is essential for virus entry to host cell. Results: Acteoside (11) showed the most potent in silico inhibition with an additional merit, over hesperidin (16), of not binding to angiotensin-converting enzyme 2 with well proven in vivo pulmonary protective role in acute lung injury, followed by chrysoeriol-7-O-ß-glucopyranoside (12) and luteolin-7-O-ß-glucopyranoside (14). Conclusion: Phytoligands (11, 12 and 14) were posed as promising candidates with potential prophylactic action against COVID-19. These phytoligands were prioritized for further biological experimentation because of their acceptable predicted ADME and drug-likeness parameters. Moreover, they could aid in developing multitargeted strategy for better management of COVID-19 using phytomedicines.

Over a century since ephedrine discovery: an updated revisit to its pharmacological aspects, functionality and toxicity in comparison to its herbal extracts.

Ephedrine, a sympathomimetic amine that exhibits several adrenaline actions, is a plant alkaloid that is a common ingredient in several cold, asthma and narcolepsy treatment preparations, and in obesity management and sport medicine. Its principal action mechanism relies on its direct adrenergic actions as well as indirect role that involves the release of epinephrine and norepinephrine, thus increasing the activity of epinephrine and norepinephrine at the postsynaptic α and ß receptors. Nevertheless, its serious side effects, including stroke, heart attack, drug abuse and interactions, have never been comprehensively reviewed. We conducted a systematic review of data on ephedrine, including its occurrence in functional foods, pharmacological aspects, metabolism, pharmaco/toxicokinetics and clinical features. Furthermore, a review of ephedrine natural structural analogues with regards to their differential adrenergic receptor binding affinities, food interaction, and their impact on the pharmacokinetics and effects relative to ephedrine are presented for the first time, and in comparison to its action when present in herbs.

Profiling of Primary Metabolites and Volatile Determinants in Mahlab Cherry (Prunus mahaleb L.) Seeds in the Context of Its Different Varieties and Roasting as Analyzed Using Chemometric Tools.

Mahlab cherry (Prunus mahaleb L.) is a plant native to the Mediterranean basin and Eastern Europe, with several health benefits and culinary uses. We explored the compositional heterogeneity in the aroma profile and nutrients of three P. mahaleb seeds in the context of its cultivar type, i.e., white and red, and in response to roasting. A holistic untargeted metabolomics approach was employed for the first time using solid-phase microextraction (SPME-GC-MS) profiles of seed volatiles and primary metabolites coupled with chemometrics. Around 65 peaks belonging to sugars, fatty acids, esters and organic acids were identified by GC-MS. White mahlab from Egypt is rich in fatty acids, e.g., oleic and α-linolenic acids. Some acyl esters, e.g., glycerylmonostearate and n-butylcaprylate, characterized mahlab cultivars from various origins. A total of 135 volatiles were identified, with organic acids and aldehydes the most abundant. Aldehydes were the most discriminatory in seed origin and in accounting for its distinct aroma. Several roasting indices were identified, viz. 1-octanol, γ-caprolactone and isomintlactone. A direct relationship between furans and fatty acids was rationalized by cyclic transformation of the latter into furan derivatives. This study provides the first chemical evidence supporting the nutritional and flavor determinants of mahlab seeds, suggesting novel uses as a functional food.

Dual prophylactic/therapeutic potential of date seed, and nigella and olive oils-based nutraceutical formulation in rats with experimentally-induced Alzheimer's disease: A mechanistic insight.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder with a multifactorial etiology and significantly increasing incidence during the last decade. Hence, developing an effective therapy is crucial for public health. The current study aimed to examine the dual prophylactic/therapeutic potential of a nutraceutical formula based on aqueous extract of roasted date seeds, and nigella and virgin-olive oils against experimentally-induced Alzheimer's disease in rats. Alzheimer's disease-like pathology was induced in male Wistar rats using oral CuSO4 (200 mg/Kg/day for two months). The nutraceutical formula was given orally to experimental animals (10 mL/kg/d) for 14 days before (as prophylaxis) and after Alzheimer's disease induction and its therapeutic effect in both cases is tested in comparison to donepezil (0.5 mg/kg/d). The nutraceutical formula was found to ameliorate the CuSO4-induced neuronal damage and regenerate the affected hippocampus tissue and significantly improvemed in learning ability. The formula was also effective in decreasing brain amyloid-ß, tau protein, TNF-α level, iNOS level in hippocampus, oxidative stress level, and inhibiting acetylcholinesterase activity and expression in brain and hippocampus, respectively. Further, an increase in GSH levels, activities of SOD, and GST and levels of hippocampus ADAM 17 and brain phospholipids was observed. In conclusion, the studied nutraceutical formula is proved to be effective in ameliorating Alzheimer's neurodegenerative progression with added-prophylactic potential.

Sweet and bitter oranges: An updated comparative review of their bioactives, nutrition, food quality, therapeutic merits and biowaste valorization practices.

Sweet and bitter oranges are two of the most commercially-important fruit with a total world production of 75.4 Mt, well-recognized for their unique sensory characters in addition to multiple nutritive and therapeutic attributes due to their highly-valued bioactive ingredients. Hence, their differential qualitative/quantitative phytochemical make-ups are presented for better utilization as therapeutic agents. Sweet orange exhibits therapeutic applications as being effective anti-diabetic, anti-obesity, and hypocholesterolemic agents. Whereas, for anti-osteoporotic products and intestinal dysbiosis treatment, bitter orange is more preferred. Moreover, the review recapitulates on different valorization practices of citrus bio-wastes and utilization of their bioactives as therapeutic agents and in functional food industry. Sweet orange waste functions as a fat replacer and preservative to increase food shelf life with better organoleptic attributes than bitter orange. The detailed action mechanism and safety of Citrus bioactives, as well as processing technologies to further improve its effects are posed as future research perspectives.

Chemometrics based GC-MS aroma profiling for revealing freshness, origin and roasting indices in saffron spice and its adulteration.

Saffron, stigmas of Crocus sativus, is one of the most precious spices used as food colorant and flavoring agent. Due to its scarce source and high cost, it is liable to fraudulent admixture with allied plants "safflower and calendula". In this study, gas chromatography-mass spectrometry (GC-MS) was employed to determine authenticity, adulterants detection, and to assess the roasting impact on its aroma. A total of 93 volatiles were identified belonging to different classes viz. aldehydes, alcohols, ketones, aliphatic hydrocarbons, aromatics, mono-and sesquiterpenes, oxides/ethers and pyrans/furans. Principle component analysis (PCA) identified safranal and 2-caren-10-al as discriminatory volatile markers of saffron from its allied flowers, later found enriched in estragole, ß-caryophyllene and eugenol. PCA model also revealed markers for freshly dried versus long-stored saffron, with ketoisophorone as freshness marker versus safranal as an ageing indicator. Safranal was further identified as a marker to distinguish saffron from safflower, whereas calendula aroma was predominated by monoterpene hydrocarbons.

Antiulcer activity of Cyperus alternifolius in relation to its UPLC-MS metabolite fingerprint: A mechanistic study.

BACKGROUND: Gastric ulcer is one of the main prevalent gastrointestinal multi-etiological disorders with many associated complications and adverse effects. Our aim was to develop safer antiulcer therapies based on methanol or ethyl acetate extracts of tubers and aerial parts from Cyperus alternifolius. METHODS: Gastric ulceration was experimentally generated by administration of single oral doses of indomethacin (30 mg/kg) to fasted rats. The animals received methanol or ethyl acetate extracts of C. alternifolius tuber and methanol or ethyl acetate extracts of aerial parts at two dose levels (50 or 100 mg/kg). Ranitidine (50 mg/kg) was used as standard anti-ulcer drug. After 4 h, the ulcer number and the total ulcer score were determined and TNF-α was assessed. Also, pathological and histochemical examination for gastric mucosa were performed. The metabolome heterogeneity of the different extracts was explored using (UPLC-MS) aided by supervised pattern recognition, i.e., orthogonal partial least squares discriminate analysis (OPLS-DA). A second OPLS-DA model was employed to link the UPLC-MS derived metabolome of the different extracts to their antiulcer activity to identify activity mediating metabolites. RESULTS: The extracts significantly reduced ulcer number, total ulcer score and TNF-α content in the stomach. Methanol or ethyl acetate extracts of tubers were most effective even more than ranitidine. In parallel, the histopathological examination showed an improvement of damaged mucosa. A high PAS reaction was observed in the treated groups indicating a relieve of the mucosal layer. A mechanistic clue of the C. alternifolius antiulcer potential was provided by the identification of its bioactive compounds using OPLS-DA. Both methanol extracts of tubers and aerial parts were more enriched in phenolic acids. The ethyl acetate extract of the aerial part was more abundant in two aldehydes. A mechanism of action was postulated based on their reported actions viz. α-carbonic anhydrase inhibition, anti-inflammatory and analgesic activity by its antioxidant activity and downregulation of several inflammatory mediators. CONCLUSION: This is the first study to report on the antiulcer activity of C. alternifolius tubers with identification of the key bioactive compounds and the mode of action. Future phytochemical and biological evaluation of the identified bioactive compounds are needed to confirm the plant tubers as safer alternative or adjunct therapy compared to conventional antiulcer drugs.

UPLC-MS metabolome based classification of Lupinus and Lens seeds: A prospect for phyto-equivalency of its different accessions.

Fabaceae is well-known for its seed nutritious and bioactive composition as exemplified by Lupinus and Lens. Developing efficient analytical approaches for profiling their bioactive matrix is a prerequisite to provide proof for their health benefits or nutritive traits. Eight Lupinus and Lens seed accessions were subjected to liquid chromatography-mass spectrometry (UPLC-MS)-based metabolomic study, which identified 66 metabolites, viz. flavonoids, alkaloids, saponins, phenolics, fatty acids and sphingolipids. Chemometric tools were explored to assess heterogeneity across the two genera leading to elucidation of the species-most enriched and differential metabolites. The two dark-colored lentil cultivars are identified as the richest source of functional foods with presumed therapeutic benefits; however, Lupinus hispanicus was proved to be the most nutritive accession. To our knowledge, this study provides the first UPLC-MS-based comparative metabolite profiling of Lupinus and Lens seeds. This platform was also able to discern metabolites diversity at the intraspecific level among Lupinus species and Lens cultivars.

Metabolome based classification of artichoke leaf: A prospect for phyto-equivalency of its different leaf origins and commercial preparations.

The growing interest in assuring phytomedicines efficacy and moreover the increase in requirements for its safety drive the development of analytical methods for its quality control assurance. Herein, we present a nuclear magnetic resonance (NMR) fingerprinting approach of artichoke leaf material from different origins and in its commercial preparations. Under optimized conditions, we were able to simultaneously identify 23 metabolites including sugars, amino and organic acids, sesquiterpene lactones, flavones, cinnamates, inulin, fatty acids and nitrogenous bases. Principal component analysis (PCA) was used to reveal for differences among artichoke specimens. PCA score plot derived from the aromatic region (5-10 ppm) provided better classification model than that of full scan (0-10 ppm), and revealing for enrichment of wild Egyptian and Tanzanian artichoke in sesquiterpene viz. aguerin B versus O-caffeoylquinic acid and luteolin abundance in cultivated leaf. PCA analysis of 3 commercial artichoke preparations showed discrimination of a silymarin-containing capsule suggesting that NMR can distinguish liver-aid herbal preparations based on its different chemical composition. Quantitative 1H NMR (qHNMR) was further employed to assess major metabolites levels and revealing for the enrichment of cultivated plants in cinnamates viz. (E)-cinnamaldehyde (1.1 mg/g) and O-caffeoyl quinic acid (15.09 mg/g,). To the best of our knowledge, this study provides the first approach utilizing NMR fingerprinting to assess for phytoequivalency among artichoke leaf and in its preparations.

Gas Chromatography/Mass Spectrometry-Based Metabolite Profiling of Nutrients and Antinutrients in Eight Lens and Lupinus Seeds (Fabaceae).

Lens culinaris and several Lupinus species are two legumes regarded as potential protein resources aside from their richness in phytochemicals. Consequently, characterization of their metabolite composition seems warranted to be considered as a sustainable commercial functional food. This study presents a discriminatory holistic approach for metabolite profiling in accessions of four lentil cultivars and four Lupinus species via gas chromatography/mass spectrometry. A total of 107 metabolites were identified, encompassing organic and amino acids, sugars, and sterols, along with antinutrients, viz., alkaloids and sugar phosphates. Among the examined specimens, four nutritionally valuable accessions ought to be prioritized for future breeding to include Lupinus hispanicus, enriched in organic ( ca. 11.7%) and amino acids ( ca. 5%), and Lupinus angustifolius, rich in sucrose ( ca. 40%), along with two dark-colored lentil cultivars 'verte du Puy' and 'Black Beluga' enriched in peptides. Antinutrient chemicals were observed in Lupinus polyphyllus, owing to its high alkaloid content. Several species-specific markers were also revealed using multivariate data analyses.