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2022
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, , 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.

2021
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 InternationalFood Reviews International: Taylor & Francis, pp. 1 - 23, 2021. AbstractWebsite

ABSTRACTDate 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.

Otify, A. M., A. - M. M. Hammam, and M. Aly Farag, "Phoenix dactylifera L. date tree pollen fertility effects on female rats in relation to its UPLC-MS profile via a biochemometric approach.", Steroids, vol. 173, pp. 108888, 2021. Abstract

Date palm (Phoenix dactylifera L.) is among the most ancient cultivated crops, of special value owing to its fruits high nutritive and economic benefits. Asides, date palm pollen is a high energy material that has been used traditionally used for fertility enhancement. In this study, effects of date palm pollen crude extract and its fractions viz., petroleum ether, methylene chloride, ethyl acetate and n-butanol on the female reproductive system were evaluated for the first time in relation to its metabolite fingerprint. Fertility activity was evaluated in immature female rats by assessing their FSH-, LH- and estrogen- activities. To pinpoint active hormonal agents in crude pollen extract and fractions, UPLC- MS analysis was employed for metabolites profiling, and in correlation to extract/fraction bioassays using multivariate OPLS analysis. Results revealed that both polar n-butanol and non-polar petroleum ether fractions exhibited the strongest activities; with a significant increase in FSH (25.7 mIU/ml in n-butanol group), estradiol (414.7 pg/ml in petroleum ether group) and progesterone levels (122.4 pg/ml in n-butanol group). Correlation between UPLC-MS and fraction bioassays was attempted using multivariate OPLS analysis to reveal for bioactive hits in these fractions. This study provides the first report on the fertility effect of date palm pollen in female rats and in relation to its metabolite fingerprint.

2019
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 : Official journal of the Metabolomic Society, vol. 15, issue 9, pp. 119, 2019. Abstract

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.

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 (Basel, Switzerland), vol. 24, issue 18, 2019. Abstract

Interest in developing coffee substitutes is on the rise, to minimizing its health side effects. In the Middle East, date palm ( 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 LD, 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.

2016
Farag, M. A., A. Otify, A. Porzel, C. G. Michel, A. Elsayed, and L. A. Wessjohann, Comparative metabolite profiling and fingerprinting of genus Passiflora leaves using a multiplex approach of UPLC-MS and NMR analyzed by chemometric tools, , vol. 408, issue 12, pp. 3125 - 3143, 2016. AbstractWebsite

Passiflora incarnata as well as some other Passiflora species are reported to possess anxiolytic and sedative activity and to treat various CNS disorders. The medicinal use of only a few Passiflora species has been scientifically verified. There are over 400 species in the Passiflora genus worldwide, most of which have been little characterized in terms of phytochemical or pharmacological properties. Herein, large-scale multi-targeted metabolic profiling and fingerprinting techniques were utilized to help gain a broader insight into Passiflora species leaves’ chemical composition. Nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS) spectra of extracted components derived from 17 Passiflora accessions and from different geographical origins were analyzed using multivariate data analyses. A total of 78 metabolites were tentatively identified, that is, 20 C-flavonoids, 8 O-flavonoids, 21 C, O-flavonoids, 2 cyanogenic glycosides, and 23 fatty acid conjugates, of which several flavonoid conjugates are for the first time to be reported in Passiflora spp. To the best of our knowledge, this study provides the most complete map for secondary metabolite distribution within that genus. Major signals in 1H-NMR and MS spectra contributing to species discrimination were assigned to those of C-flavonoids including isovitexin-2″-O-xyloside, luteolin-C-deoxyhexoside-O-hexoside, schaftoside, isovitexin, and isoorientin. P. incarnata was found most enriched in C-flavonoids, justifying its use as an official drug within that genus. Compared to NMR, LC-MS was found more effective in sample classification based on genetic and/ or geographical origin as revealed from derived multivariate data analyses. Novel insight on metabolite candidates to mediate for Passiflora CNS sedative effects is also presented.

2015
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 & Function, vol. 6, issue 12: The 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 [gamma]-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.