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Abstract Summary Phoenix roebelenii O’Brien is among 202 genera and 2300-2700 species of the family Arecaceae being largely distributed in the tropics and subtropics. Both genus and species are widely spread and cultivated in Egypt. Genus phoenix has been widely used in folk medicine for the treatment of various infectious diseases, atherosclerosis, diabetes, hypertension and cancer. Members of the family Arecaceae are characterized by different classes of phenolic compounds, C-glycosylflavones, luteolin and quercitin glycosides. Phenolic compounds are currently receiving much interest as a potential source of therapeutic agents for clinical use because they serve as antioxidant. Oxidative stress is one of the common denominator in pathogenesis of chronic disease. Our study was focusing on isolation and identification of phenolics in P. roebelenii, in addition to evaluation of certain biological activities in an effort to report what is considered the first phytochemical and biological reports on this species worldwide. The work carried out in the present study was divided into six main parts: 1. Literature survey on Phoenix species belonging to family Arecaceae Literature survey revealed that Phoenix roebelenii O’Brien has not been subjected to any phytochemical nor biological investigation before. 2. Taxonomical and morphological feature of genus and species of P. roebelenii O’Brien 3. DNA profiling The extracted DNA of Phoenix roebelenii O’Brien was amplified using ten decamer primers to detect its genetic pattern. The ten primers had successfully directed the amplification of a genome-specific fingerprint of DNA fragments. The ten primers (A15, A04, A17, A18, C01, C19, C07, D05, G19 and M02) of arbitrary sequences generated 91 fragments. The ten primers had produced multiple band profiles with a number of amplified DNA fragments ranging from 15 when C01 was used. Whereas, the least number of fragments was 4 being produced by D05. Therefore primer C01 was the best sequence for dominating Phoenix roebelenii O’Brien cultivated in Egypt. 4. Preliminary phytochemical investigation 4.1 Phytochemical screening of the leaves and fruits of Phoenix roebelenii O’Brien Phytochemical screening revealed the presence of carbohydrates and/or glycosides, sterols and/or triterpens, tannins and saponins in both leaves and fruit of P. roebelenii O’Brien. Phenolics and/or flavonoids and coumarins were detected in the leaves of P. roebelenii O’Brien while found in trace amounts in the fruit. Anthraquinone and alkaloids were not detected in both leaves and fruit. 4.2 Determination of Macro and Micro-elements: Micro elements (Pb, Fe, Mn, Zn, Cu, Co, Cr, Ni and Cd) were determined in the total ash of both leaves and fruits of P. roebelenii adopting atomic absorption method, while Macro elements (P, K, Na and Ca) were determined adopting flame photometric method, the main findings in this study were the high content of Fe and Zn in both leaves and fruits and the absence of Pb and Cd elements in the leaves of P. roebelenii O’Brien. Summary 151 5. Chemical investigation 5.1. Quantitative estimation of the phenolic content The total phenol content of the leaves and fruit of P. roebelenii O’Brien were determined using Folin-Ciocalteau method. It was found to be 64.3 μg GAE/mg for leaves and 7.88 μg GAE/mg for fruits. The total flavonoid contents of the leaves and fruit of P. roebelenii O’Brien were determined using spectrophotometric method. It was found to be 149.4 μg RE/mg for leaves and 8.66 μg RE/mg for fruits. 5.2. Preparation of the extracts for chemical investigation Aqueous methanol extract of the leaves of P. roebelenii O’Brien was prepared by repeated percolation on hot with 70% methanol, then defatted with chloroform. The defatted residue was extracted with methanol and followed by water (three times for each solvent). 5.3. chromatographic investigation 2 Dimension PC investigation of 70% Methanol extract revealed the presence of 10 major phenolic and/or flavonoids compounds. 5.4. Isolation and purification Isolation of the phenolic compounds of P. roebelenii O’Brien included polyamide column chromatographic investigation. Separation of individual flavonoids and phenolic acids was performed by subsequent PPC from column fractions, repeated purification of these individuals by rechromatography on sephadex LH-20 column and establishment of their purity by paper chromatography investigation. 5.5. Identification and structure elucidation For the structural elucidation of the isolated phenolic compounds, the required information was obtained through application of chemical degradation methods (acid hydrolysis and ferric chloride degradation), spectroscopic methods (UV, 1HNMR and 13C-NMR) and spectrometric techniques of analysis (EI/MS and ESI/MS). Phenolic compounds isolated from P. roebelenii O’Brien Ten compounds (P1 – P10) were isolated and identified for the first time from the leaves of P. roebelenii O’Brien. To our best of knowledge compounds P3 a |