Pecio, Ł., A. M. Otify, F. R. Saber, Y. A. El-Amier, M. E. Shalaby, S. Kozachok, A. K. Elmotayam, Ł. Świątek, A. Skiba, and K. Skalicka-Woźniak, "Iphiona mucronata (Forssk.) Asch. & Schweinf. A Comprehensive Phytochemical Study via UPLC-Q-TOF-MS in the Context of the Embryo- and Cytotoxicity Profiles", Molecules, vol. 27, issue 21, 2022. Abstract

Iphiona mucronata (Family Asteraceae) is widely distributed in the Eastern desert of Egypt. It is a promising plant material for phytochemical analysis and pharmacologic studies, and so far, its specific metabolites and biological activity have not yet been thoroughly investigated. Herein, we report on the detailed phytochemical study using UPLC-Q-TOF-MS approach. This analysis allowed the putative annotation of 48 metabolites belonging to various phytochemical classes, including mostly sesquiterpenes, flavonoids, and phenolic acids. Further, zebrafish embryotoxicity has been carried out, where 100 µg/mL extract incubated for 72 h resulted in a slow touch response of the 10 examined larvae, which might be taken as a sign of a disturbed peripheral nervous system. Results of in vitro testing indicate moderate cytotoxicity towards VERO, FaDu, and HeLa cells with CC50 values between 91.6 and 101.7 µg/mL. However, selective antineoplastic activity in RKO cells with CC50 of 54.5 µg/mL was observed. To the best of our knowledge, this is the first comprehensive profile of I. mucronata secondary metabolites that provides chemical-based evidence for its biological effects. A further investigation should be carried out to precisely define the underlying mechanisms of toxicity.

Ibrahim, R. M., B. M. Eltanany, L. Pont, F. Benavente, S. A. ElBanna, and A. M. Otify, "Unveiling the functional components and antivirulence activity of mustard leaves using an LC-MS/MS, molecular networking, and multivariate data analysis integrated approach", Food Research International, vol. 168, 2023. Abstract
Otify, A. M., A. Serag, A. Porzel, L. A. Wessjohann, and M. A. Farag, "NMR Metabolome-Based Classification of Cymbopogon Species: a Prospect for Phyto-equivalency of its Different Accessions Using Chemometric Tools", Food Analytical Methods, vol. 15, no. 8: Springer, pp. 2095 – 2106, 2022. AbstractWebsite

Cymbopogon species are widely distributed worldwide and known for their high essential oil content with potential commercial and medicinal benefits justifying for their inclusion in food and cosmetics. Most species received scant characterization regarding their full complement of bioactive constituents necessary to explain their medicinal activities. In this study, the metabolite profiles of 5 Cymbopogon species, C. citratus, C. flexuosus, C. procerus, C. martini, and C. nardus, were characterized via NMR-based metabolomics. The results of 13 shoot accessions revealed the identification and quantification of 23 primary and secondary metabolites belonging to various compound classes. Multivariate analyses were used for species classification, though found not successful in discrimination based on geographical origin. Nevertheless, C. citratus was found particularly enriched in neral, geranial, (E)-aconitic acid, isoorientin, and caffeic acid as the major characterizing metabolites compared to other species, while an unknown apigenin derivative appeared to discriminate C. martini. The high essential oil and phenolic content in C. citratus emphasizes its strong antioxidant activity, whereas (E)-aconitic acid accounts for its traditional use as insecticide. This study affords the first insight into metabolite compositional differences among Cymbopogon species. Moreover, antimicrobial, insecticidal, antidiabetic, and antioxidant compounds were identified that can be utilized as biomarkers for species authentication. © 2022, The Author(s).

Otify, A. M., A. M. El-Sayed, C. G. Michel, and M. A. Farag, "Metabolites profiling of date palm (Phoenix dactylifera L.) commercial by-products (pits and pollen) in relation to its antioxidant effect: a multiplex approach of MS and NMR metabolomics", Metabolomics, vol. 15, no. 9: Springer New York LLC, 2019. AbstractWebsite

Introduction: Phoenix dactylifera L. (date palm) is one of the most valued crops worldwide for its economical and nutraceutical applications of its date fruit (pericarp). Currently date pits, considered as a waste product, is employed as coffee substitute post roasting. Whereas, pollen represents another valuable by-product used as a dietary supplement. Objectives: In this study, a large-scale comparative metabolomics approach was performed for the first characterization and standardization of date palm by-products viz., date pits and pollen. Moreover, roasting impact on date pit metabolite composition was also assessed. Methods: Metabolites profiling of pits and pollen was determined via a multiplex approach of UPLC–MS and NMR, coupled to multivariate analysis, in relation to its antioxidant activities. Results: Chemical analyses led to the identification of 67 metabolites viz., phenolic acids, flavonols, fatty acids, sphingolipids, steroids and saponins of which 10 are first time to be reported. The enrichment of steroids in date pollen accounts for its fertility promoting properties, whereas date pit was found a rich source for antioxidant polyphenols using metabolomics. © 2019, Springer Science+Business Media, LLC, part of Springer Nature.

Otify, A. M., S. A. Elbanna, B. M. Eltanany, L. Pont, F. Benavente, and R. M. Ibrahim, "A comprehensive analytical framework integrating liquid chromatography-tandem mass spectrometry metabolomics with chemometrics for metabolite profiling of lettuce varieties and discovery of antibacterial agents", Food Research International, vol. 172: Elsevier Ltd, 2023. AbstractWebsite

This study comprehensively characterized the metabolite profiles of six lettuce varieties and established the correlation between the elucidated profiles and their antivirulence effects. A total of 195 metabolites were annotated using LC-QTOF-MS/MS metabolomics assisted by molecular networking and integrated with chemometrics. Red varieties (red longifolia and lolla rosa) demonstrated higher chlorogenic and chicoric acids suggesting their antioxidant properties. In parallel, amino acids and disaccharides were enriched in romaine longifolia rationalizing its palatable taste and nutritional potential, while crispa, capitata, and lolla bionda presented a high β-carboline alkaloid content. The antibacterial and antihemolytic potential of all varieties against methicillin-sensitive and methicillin-resistant Staphylococcus aureus was assessed and validated by prominent downregulation of α-hemolysin transcriptional levels in both strains. Moreover, correlation analysis revealed sesquiterpenes, β-carboline alkaloids, amino acids, and oxy-fatty acids as the main bioactives. Results emphasize lettuce significance as a functional food and nutraceutical source, and highlight varieties naturally rich in antibacterial agents to adapt breeding programs. © 2023 Elsevier Ltd

Farag, M. A., A. M. Otify, A. M. El-Sayed, C. G. Michel, S. A. ElShebiney, A. Ehrlich, and L. A. Wessjohann, "Sensory metabolite profiling in a date pit based coffee substitute and in response to roasting as analyzed via mass spectrometry based metabolomics", Molecules, vol. 24, no. 18: MDPI AG, 2019. AbstractWebsite

Interest in developing coffee substitutes is on the rise, to minimizing its health side effects. In the Middle East, date palm (Phoenix dactylifera L.) pits are often used as a coffee substitute post roasting. In this study, commercially-roasted date pit products, along with unroasted and home-prepared roasted date pits, were subjected to analyses for their metabolite composition, and neuropharmacological evaluation in mice. Headspace SPME-GCMS and GCMS post silylation were employed for characterizing its volatile and non-volatile metabolite profile. For comparison to roasted coffee, coffee product was also included. There is evidence that some commercial date pit products appear to contain undeclared additives. SPME headspace analysis revealed the abundance of furans, pyrans, terpenoids and sulfur compounds in roasted date pits, whereas pyrroles and caffeine were absent. GCMS-post silylation employed for primary metabolite profiling revealed fatty acids’ enrichment in roasted pits versus sugars’ abundance in coffee. Biological investigations affirmed that date pit showed safer margin than coffee from its LD50, albeit it exhibits no CNS stimulant properties. This study provides the first insight into the roasting impact on the date pit through its metabolome and its neuropharmacological aspects to rationalize its use as a coffee substitute. © 2019 by the authors.

Otify, A. M., R. M. Ibrahim, B. Abib, A. Laub, L. A. Wessjohann, Y. Jiang, and M. A. Farag, "Unveiling metabolome heterogeneity and new chemicals in 7 tomato varieties via multiplex approach of UHPLC-MS/MS, GC–MS, and UV–Vis in relation to antioxidant effects as analyzed using molecular networking and chemometrics", Food Chemistry, vol. 417: Elsevier Ltd, 2023. AbstractWebsite

Tomatoes show diverse phytochemical attributes that contribute to their nutritive and health values. This study comprehensively dissects primary and secondary metabolite profiles of seven tomato varieties. UHPLC-qTOF-MS assisted molecular networking was used to monitor 206 metabolites, 30 of which were first-time to be reported. Flavonoids, as valuable antioxidants, were enriched in light-colored tomatoes (golden sweet, sun gold, and yellow plum) versus high tomatoside A, an antihyperglycemic saponin, in cherry bomb and red plum varieties. UV–Vis analysis revealed similar results with a strong absorbance corresponding to rich phenolic content in light varieties. GC–MS unveiled monosaccharides as the main contributors to samples’ segregation, found abundant in San Marzano tomato accounting for its sweet flavor. Fruits also demonstrated potential antioxidant activities in correlation to their flavonoids and phospholipids. This work provides a complete map of tomatoes’ metabolome heterogeneity for future breeding programs and a comparative approach utilizing different metabolomic platforms for tomato analysis. © 2023 Elsevier Ltd

Otify, A., C. George, A. Elsayed, and M. A. Farag, "Mechanistic evidence of Passiflora edulis (Passifloraceae) anxiolytic activity in relation to its metabolite fingerprint as revealed via LC-MS and chemometrics", Food and Function, vol. 6, no. 12: Royal Society of Chemistry, pp. 3807 – 3817, 2015. AbstractWebsite

Passiflora edulis Sims F. flavicarpa along with several other plants belonging to the genus Passiflora have been reported as sedatives and for treatment or prevention of central disorders. This study evaluated the anxiolytic effect of P. edulis ethanol extract and its fractions (viz. chloroform, ethyl acetate and butanol) using the elevated plus-maze model of anxiety and assessment of γ-aminobutyric acid levels. The results revealed that butanol and chloroform extracts exhibit the strongest effect followed by ethyl acetate suggesting that a combination of different classes of metabolites is likely to mediate for P. edulis anxiolytic effect in these fractions. To further pinpoint bioactive agents in fractions, ultra-performance liquid chromatography (UPLC) coupled to high resolution qTOF-MS was used for secondary metabolite profiling. A total of 65 metabolites were characterized including O-flavonoids, C-flavonoids, cyanogenic glycosides and fatty acids. Harman type alkaloids found in P. incarnata were not detected in P. edulis ethanol extract or any of its fractions suggesting that they do not mediate for its CNS modulating effects. Multivariate data analysis (PCA) was further applied to identify metabolite markers for fractions and revealed that enrichment of C-glycoside type flavonoids in chloroform/ethyl acetate fractions versus the exclusive presence of cyanogenic glycosides in its butanol fraction. © 2015 The Royal Society of Chemistry.

Farag, M. A., M. G. Sharaf El-Din, M. A. Selim, A. I. Owis, S. F. AbouZid, A. Porzel, L. A. Wessjohann, and A. Otify, "Nuclear magnetic resonance metabolomics approach for the analysis of major legume sprouts coupled to chemometrics", Molecules, vol. 26, no. 3: MDPI AG, 2021. AbstractWebsite

Legume sprouts are a fresh nutritive source of phytochemicals of increasing attention worldwide owing to their many health benefits. Nuclear magnetic resonance (NMR) was utilized for the metabolite fingerprinting of 4 major legume sprouts, belonging to family Fabaceae, to be exploited for quality control purposes. Thirty-two metabolites were identified belonging to different classes, i.e., fatty acids, sugars, amino acids, nucleobases, organic acids, sterols, alkaloids, and isoflavonoids. Quantitative NMR was employed for assessing the major identified metabolite levels and multivariate data analysis was utilized to assess metabolome heterogeneity among sprout samples. Isoflavones were detected exclusively in Cicer sprouts, whereas Trigonella was characterized by 4-hydroxyisoleucine. Vicia sprouts were distinguished from other legume sprouts by the presence of L-Dopa versus acetate abundance in Lens. A common alkaloid in all sprouts was trigonelline, detected at 8–25 µg/mg, suggesting its potential role in legume seeds’ germination. Trigonelline was found at highest levels in Trigonella sprouts. The aromatic NMR region data (δ 11.0–5.0 ppm) provided a better classification power than the full range (δ 11.0–0.0 ppm) as sprout variations mostly originated from secondary metabolites, which can serve as chemotaxonomic markers. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Farag, M. A., A. Otify, and M. H. Baky, "Phoenix Dactylifera L. Date Fruit By-products Outgoing and Potential Novel Trends of Phytochemical, Nutritive and Medicinal Merits", Food Reviews International, vol. 39, no. 1: Taylor and Francis Ltd., pp. 488 – 510, 2023. AbstractWebsite

Date palm (Phoenix dactylifera L., Arecaceae) is a valuable fruit tree for many populations living in the Arabian Peninsula, North Africa, and the Middle East, and to play significant role in its economy. Asides from its well-known edible fruit, date palm tree also provides numerous by-products which have yet to be fully capitalized upon commercially. All parts of the date palm tree are of value to include: seeds (pits and oil), date pollen and the leaves. Date palm seed is a by-product of the fruit industry being discarded, and used as animal feed or for human consumption as non-caffeinated coffee. Date pollen is used traditionally for the treatment of infertility. This review aims to introduce a holistic overview on the phytochemical composition, nutritional, economical and potential health benefits of date by-products. Prioritization of date palm by-products for a desired added value or production of certain economically valued products is presented. © 2021 Taylor & Francis.