Olive Oil Phenols

Olive oils contain numerous substances that have a beneficial role in human health. Phenols are natural compounds that are present in extra-virgin olive oil (EVOO), and they are produced at the malaxation step of the olive oil production. The four most abundant phenols in EVOO are oleocanthal, oleacein, ligstroside aglycon, and oleuropein aglycon. These phenols exhibit significant biological effects in many diseases, participating in various cellular and biochemical processes. Oleocanthal can protect and prevent against the Alzheimer disease, demonstrates acute antiplatelet effects, which has a vital role against cancer, and can act like ibuprofen. Oleacein has antioxidant and anti-inflammatory activities and helps against atherosclerosis. Moreover, it acts as an antiaging factor and as a 5-lipoxygenase inhibitor. Ligstroside aglycon implicates to mechanisms against breast cancer, while oleuropein aglycon shows activities against the Alzheimer disease and breast cancer

Fatty Acids Derived from Royal Jelly Are Modulators of Estrogen Receptor Functions

Royal jelly (RJ) excreted by honeybees and used as a nutritional and medicinal agent has estrogen-like effects, yet the compounds mediating these effects remain unidentified. The possible effects of three RJ fatty acids (FAs) (10-hydroxy-2-decenoic-10H2DA, 3,10-dihydroxydecanoic-3,10DDA, sebacic acid-SA) on estrogen signaling was investigated in various cellular systems. In MCF-7 cells, FAs, in absence of estradiol (E2), modulated the estrogen receptor (ER) recruitment to the pS2 promoter and pS2 mRNA levels via only ERβ but not ERα, while in presence of E2 FAs modulated both ERβ and ERα. Moreover, in presence of FAs, the E2-induced recruitment of the EAB1 co-activator peptide to ERα is masked and the E2-induced estrogen response element (ERE)-mediated transactivation is inhibited. In HeLa cells, in absence of E2, FAs inhibited the ERE-mediated transactivation by ERβ but not ERα, while in presence of E2, FAs inhibited ERE-activity by both ERβ and ERα. Molecular modeling revealed favorable binding of FAs to ERα at the co-activator-binding site, while binding assays showed that FAs did not bind to the ligand-binding pocket of ERα or ERβ. In KS483 osteoblasts, FAs, like E2, induced mineralization via an ER-dependent way. Our data propose a possible molecular mechanism for the estrogenic activities of RJ's components which, although structurally entirely different from E2, mediate estrogen signaling, at least in part, by modulating the recruitment of ERα, ERβ and co-activators to target genes

Chemical analysis and biological activities of honey, royal jelly, propolis and bee plants

In the 1est chapter, the dichloromethane and methanol extracts of royal jelly from Greece were studied. Twenty six compounds were isolated and identified. Among them, seventeen compounds are reported for the first time as royal jelly constituents, while seven of them are isolated for the first time as natural products. The seven new compounds were fatty acid derivatives: 10-acetoxydecanoic acid (31), trans-10-acetoxydec-2-enoic acid (32), 11-oxododecanoic acid (40), (11S)-hydroxydodecanoic acid (43), (10R,11R)- dihydroxydodecanoic acid (44), 3,11-dihydroxydodecanoic acid (46) and (11S),12- dihydroxydodecanoic acid (47). The studied sample and the isolated compounds were tested for their antimicrobial, antioxidant, estrogenic, antidiabetic, wound-healing and cytotoxic activity. From all of these activities the most interesting were the antimicrobial, estrogenic and antidiabetic. OH COOCH3 methyl paraben OH OCH3 CH2CH2COOH 3-(4-Hydroxy-3-methoxyphenyl)propionic acid O OH OH 11(S)-hydroxydodecanoic acid (43) O OH O OH sebacic acid (7) 10-hydroxy-2-decenoic acid (1) O OH HO 10-acetoxy-decanoic acid (31) O O OH CH3 O 10-acetoxy-2-decenoic acid (32) O O OH CH3 O OH COCH3 OCH3 acetovanillone (34) OH COCH3 p-hydroxyacetophenone O O δ-decalactone (35) O O δ-octalactone (36) octanoic acid (39) O OH HO O 9-hydroxy-2-nonanone (41) New natural product New natural product New natural product (30) (42) (33) 8-Hydroxyoctanoic acid (9) O OH HO 3-Hydroxydecanoic acid methyl ester (37) O OH OCH3 HO O OH OH 3,10-dihydroxydecanoic acid (16) O O OH 11-οxododecanoic acid (40) 3,11-dihydroxydodecanoic acid (46) O OH OH OH HO O OH 10-hydroxydecanoic acid (2) Νew natural product New natural product New natural product New natural product 3-hydroxydodecanedioic acid (45) O OH OH O OH HO 24-methylene cholesterol (22) 11(S),12-dihydroxydodecanoic acid (47) OH O HO OH OH O OH OH HO H H H β-sitosterol (23) HO H H isofucosterol (24) 10(R),11(R)-dihydroxydodecanoic acid (44) COOH benzoic acid (38) In the 2nd chapter, the n-butanol extract of Greek propolis was studied. One new 2,3 dihydroflavone derivative, 7-prenylstrobopinin (96), and 25 known diterpenes and phenolic compounds were isolated and identified. It is the first time that diterpenes are isolated from propolis of European origin, while six of the known compounds are reported as propolis constituents for the first time. The studied sample and the isolated compounds were tested for their antimicrobial, antioxidant, and cytotoxic activity. From all of these activities the most interesting were the antimicrobial and antioxidant activity. CH3O O OH O HO O OH O CH3O O OH O OH O O OH O OH COOH OH COOH OH COOH OHC CH3 CH2OH pinocebrin (55) sakuranetin (101) 7- prenyl-pinocebrin (97) P-coumaric acid (66) caffeic acid (67) isoagatholal (100) pinostrobin (53) OH H benzoic acid (38) trans- totarol (95) 1' 9 7 6 5 2 3'' 2'' 1'' B A 7- prenyl-strobopinin (96) O O OH O H3C New natural product HOH2C CH3 CH2 OH HOOC CH3 CH2 OH 13-13-epi-torulosol (104) epi-cupressic acid (103) HO O OH O OH HO O O O OH CH3 HO O OH O Chrysin (48) CH2OH H3C CH3 copalol (98) pinobanksin (54) 5-methyl-Pinobanksin (102) O O O O O O benzyl cinnamate (107) benzyl benzoate (109) cinnamyl cinnamate (76) O O (CH2)3 CH3 OH ferruginol (106) butyl cinnamate (108) vanillyl butylate (110) CH3 (CH2)2 Ο Ο OH OCH3 CH2OH HOOC CH3 OHC CH3 OH HOH2C CH3 CH2OH Isocupressic acid (59) Agathadiol (105) 13- epi-torulosal (99) In the 3nd chapter were studied: a mono-floral honey sample coming from orange trees and a honey sample coming from thymus. From these two kinds of honey were isolated hydroxymethylfurfural and p-benzoic acid. From the honey sample coming from orange trees was isolated (+) 8-hydroxylinalool. In this study is suggested that (+) 8- hydroxylinalool could be used as a chemical marker of honey samples coming from orange trees. 3-Hydroxy-4-phenyl-2-butanone was isolated from honey of thymus and it is the first time that this constituent is isolated from honey. Through the antimicrobial screening, most of the tested honeys as well the characteristic 3- hydroxy-4-phenyl-2-butanone and (+) 8-hydroxylinalool, proved to be significantly active against all tested microorganisms. Additionally the honey samples exhibited interesting antioxidant activity. O HOCH2 CHO OH hydroxymethylfurfural (111) HO O p-hydroxybenzoic acid (20) CH3 O OH OH OH 3-hydroxy-Στο 1ο κεφάλαιο μελετήθηκε το διχλωρομεθανικό και το μεθανολικό εκχύλισμα ενός δείγματος Ελληνικού βασιλικού πολτού. Απομονώθηκαν και ταυτοποιήθηκαν συνολικά 26 φυσικά προϊόντα. Από το σύνολο των προϊόντων που απομονώθηκαν τα 17 συστατικά για πρώτη φορά ταυτοποιούνται ως συστατικά βασιλικού πολτού, ενώ 7 από αυτά είναι νέα φυσικά προϊόντα. Τα 7 νέα φυσικά προϊόντα ήταν παράγωγα λιπαρών οξέων: 10- ακετοξυδεκανοϊκό οξύ (31), trans-10-ακετοξυ-2-δεκενοϊκό οξύ (32), 11- οξοδωδεκανοϊκό οξύ (40), (11S)-υδροξυδωδεκανοϊκό οξύ (43), (10R,11R)- διυδροξυδωδεκανοϊκό οξύ (44), 3,11-διυδροξυδωδεκανοϊκό οξύ (46) and (11S),12- διυδροξυδωδεκανοϊκό οξύ (47). Τα εκχυλίσματα και τα συστατικά που απομονώθηκαν, μελετήθηκαν ως προς την αντιμικροβιακή, αντιοξειδωτική, επουλωτική, κυτταροτοξική, αντιδιαβητική και οιστρογονική δράση. Ιδιαίτερο ενδιαφέρον παρουσίασαν η αντιμικροβιακή δράση, η οιστρογονική και η αντιδιαβητική δράση. OH COOCH3 OH OCH3 CH2CH2COOH 3-(4-Hydroxy-3-μεθοξυφαινυλ)προπιονικό οξύ O OH O OH σεμπακικό οξύ (7) 10-υδροξυ-2-δεκενοϊκό οξύ (1) O OH HO 10-ακετοξυ-δεκανοϊκό οξύ (31) O O OH CH3 O 10-ακετοξυ-2-δεκενοϊκό οξύ (32) O O OH CH3 O OH COCH3 OCH3 ακετοβανιλλόνη (35) OH COCH3 p-υδροξυακετοφαινόνη (33) O O O O δ-οκταλακτόνη (36) Οκτανοϊκό οξύ (39) O OH HO O 9-υδροξυ-2-νονανόνη (41) p-υδροξυβενζοϊκός μεθυλεστέρας δ-δεκαλακτόνη (35) 11(S)-υδροξυδωδεκανοϊκό οξύ (43) OH OH O Νέο φυσικό προϊόν Νέο φυσικό προϊόν Νέο φυσικό προϊόν (30) (42) (34) Περίληψη 420 8-υδροξυοκτανοϊκό οξύ (9) O OH HO μεθυλεστέρας του 3-υδροξυδεκανοϊκού οξέως (37) O OH OCH3 10(R),11(R)-διυδροξυδωδεκανοϊκό οξύ (44) HO O OH OH 3,10-διυδροξυδεκανοϊκό οξύ (16) O O OH 11-οξοδωδεκανοϊκό οξύ (40) 3,11-διυδροξυδωδεκανοϊκό οξύ (46) O OH OH OH ισοφουκοστερόλη (24) HO H H HO β-σιτοστερόλη (23) H H H OH O OH OH OH O HO OH 11(S),12-διυδροξυδωδεκανοϊκό οξύ (47) 24-μεθυλενχολεστερόλη (22) HO O OH OH O OH 3-υδροξυδωδεκανοδιοϊκό οξύ (45) Νέο φυσικό προϊόν Νέο φυσικό προϊόν Νέο φυσικό προϊόν Νέο φυσικό προϊόν 10-υδροξυδεκανοϊκό οξύ (2) HO O OH COOH βενζοϊκό οξύ (38) Στο 2ο κεφάλαιο μελετήθηκε το βουτανολικό εκχύλισμα ενός δείγματος Ελληνικής πρόπολης. Απομονώθηκαν και ταυτοποιήθηκαν 25 φυσικά προϊόντα. Από τα παραπάνω ένα φλαβονοειδές η 7-πρενυλοστρομποπινίνη είναι νέο φυσικό προϊόν, ενώ 6 συστατικά για πρώτη φορά ταυτοποιήθηκαν ως συστατικά πρόπολης από οποιοδήποτε μέρος του κόσμου. Επίσης για πρώτη φορά ανιχνεύτηκαν διτερπένια σε πρόπολη Ευρωπαϊκής προέλευσης. Τα εκχυλίσματα και τα συστατικά που απομονώθηκαν, παρουσίασαν πολύ καλή αντιμικροβιακή, κυτταροτοξική και αντιοξειδωτική δράση. CH3O O OH O HO O OH O CH3O O OH O OH O O OH O OH COOH OH COOH OH COOH OHC CH3 CH2OH πινοκεμπρίνη (55) σακουρανετίνη (101) 7- πρενυλ-πινοκεμπρίνη (97) P-υδροξυκουμαρικό οξύ (66) καφεϊκό οξυ (67) ισοαγκαθολάλη (100) πινοστρομπίνη (53) OH H βενζοϊκό οξύ (38) trans- τοταρόλη (95) 1' 9 7 6 5 2 3'' 2'' 1'' B A 7- πρενυλ-στρομποπινίνη (96) O O OH O H3C Νέο φυσικό προϊόν HOH2C CH3 CH2 OH HOOC CH3 CH2 OH 13-επι-τορουλοσόλη (104) 13-επι-κουπρεσσικό οξύ (103) HO O OH O OH HO O O O OH CH3 HO O OH O χρυσίνη (48) CH2OH H3C CH3 κοπαλόλη (98) πινομπανκσίνη (54) 5-μεθυλ..

High-Throughput 1H-Nuclear Magnetic Resonance-Based Screening for the Identification and Quantification of Heartwood Diterpenic Acids in Four Black Pine (Pinus nigra Arn.) Marginal Provenances in Greece

A high-throughput quantitative Nuclear Magnetic Resonance 1H-NMR method was developed and applied to screen the quantity of the diterpenic resin acids in the heartwood of black pine, due to the renewed scientific interest in their medicinal properties and use in various diseases treatment. The 260 samples were taken from Pinus nigra clones, selected from four provenances of the Peloponnese (Greece), participating in a 35-year-old clonal seed orchard. Total resin acids per dry heartwood weight (dhw) varied greatly, ranging from 30.05 to 424.70 mg/gdhw (average 219.98 mg/gdhw). Abietic was the predominant acid (76.77 mg/gdhw), followed by palustric acid (47.94 mg/gdhw), neoabietic acid (39.34 mg/gdhw), and pimaric acid (22.54 mg/gdhw). Dehydroabietic acid was at moderate levels (11.69 mg/gdhw), while levopimaric, isopimaric, and sandaracopimaric acids were in lower concentrations. The resin acid fraction accounted for 72.33% of the total acetone extractives. Stilbenes were presented in significant quantities (19.70%). The resin acid content was composed mainly of the abietane type resin acids (83.56%). Peloponnesian Pinus nigra heartwood was found to be the richest source of resin acids identified to date and is considered the best natural source for the production of such bioactive extracts. The results indicate a high potential for effective selection and advanced breeding of pharmaceutical and high economic value bioactive substances from Pinus nigra clones

A Method for the Rapid Measurement of Alkylresorcinols in Flour, Bread and Related Products Based on 1H qNMR

The main objectives of the current work were to investigate differences among flours from traditionally preserved Greek varieties of cereals, and especially those of wheat, and in parallel, to correlate those potential differences with the presence of bioactive natural ingredients. In this context, we developed a new, fast, and simple method for the measurement of total 5-alkylresorcinols in cereals and related foods by qNMR. Several types of flour (white or whole-grain) coming from wheat, i.e., Triticum dicoccum, T. monococcum, T. aestivum, T. durum and T. turgidum, corn, barley, rye and oat from a certified producer in Greece were used either as raw materials or for the production of bread, pasta and flakes. A small portion of the flour or the corresponding product was extracted with DMSO-d6. The liquid part was directly analyzed by NMR (400 MHz). The simplicity of the NMR spectrum of the total extract and the lack of overlapping peaks permitted the development of a high throughput quantitative method for the measurement of total bioactive alkylresorcinols in less than 15 min. Grains, whole grain flours and breads from old varieties of T. dicoccum and T.monococcum showed high contents of alkylresorcinols (455–1148 mg/Kg), while the same compounds were completely absent from white flour and the corresponding bread. The term high-phenolic flour is proposed to distinguish among flour types

Ultrahigh-pressure liquid chromatography triple-quadrupole tandem mass spectrometry quantitation of polyphenols and secoiridoids in california-style black ripe olives and dry salt-cured olives.

The chemical composition of finished table olive products is influenced by the olive variety and the processing method used to debitter or cure table olives. Herein, a rapid ultrahigh-pressure liquid chromatography triple-quadrupole tandem mass spectrometry method, using dynamic multiple reaction monitoring, was developed for the quantitation of 12 predominant phenolic and secoiridoid compounds in olive fruit, including hydroxytyrosol, oleuropein, hydroxytyrosol-4-O-glucoside, luteolin-7-O-glucoside, rutin, verbascoside, oleoside-11-methyl ester, 2,6-dimethoxy-p-benzoquinone, phenolic acids (chlorogenic and o-coumaric acids), oleuropein aglycone, and ligstroside aglycone. Levels of these compounds were measured in fresh and California-style black ripe processed Manzanilla olives and two dry salt-cured olive varieties (Mission from California and Throuba Thassos from Greece). Results indicate that the variety and debittering processing method have strong impact on the profile of phenolic and secoiridoid compounds in table olives. The dry salt-cured olives contained higher amounts of most compounds studied, especially oleuropein (1459.5 ± 100.1 μg/g), whereas California-style black ripe olives had a significant reduction or loss of these bioactive compounds (e.g., oleuropein level at 36.7 ± 3.1 μg/g)

Direct Quantitation of Phytocannabinoids by One-Dimensional 1H qNMR and Two-Dimensional 1H-1H COSY qNMR in Complex Natural Mixtures

The widespread use of phytocannabinoids or cannabis extracts as ingredients in numerous types of products, in combination with the legal restrictions on THC content, has created a need for the development of new, rapid, and universal analytical methods for their quantitation that ideally could be applied without separation and standards. Based on previously described qNMR studies, we developed an expanded 1H qNMR method and a novel 2D-COSY qNMR method for the rapid quantitation of ten major phytocannabinoids in cannabis plant extracts and cannabis-based products. The 1H qNMR method was successfully developed for the quantitation of cannabidiol (CBD), cannabidiolic acid (CBDA), cannabinol (CBN), cannabichromene (CBC), cannabichromenic acid (CBCA), cannabigerol (CBG), cannabigerolic acid (CBGA), Δ9-tetrahydrocannabinol (Δ9-THC), Δ9-tetrahydrocannabinolic acid (Δ9-THCA), Δ8-tetrahydrocannabinol (Δ8-THC), cannabielsoin (CBE), and cannabidivarin (CBDV). Moreover, cannabidivarinic acid (CBDVA) and Δ9-tetrahydrocannabivarinic acid (Δ9-THCVA) can be distinguished from CBDA and Δ9-THCA respectively, while cannabigerovarin (CBGV) and Δ8-tetrahydrocannabivarin (Δ8-THCV) present the same 1H-spectra as CBG and Δ8-THC, respectively. The COSY qNMR method was applied for the quantitation of CBD, CBDA, CBN, CBG/CBGA, and THC/THCA. The two methods were applied for the analysis of hemp plants; cannabis extracts; edible cannabis medium-chain triglycerides (MCT); and hemp seed oils and cosmetic products with cannabinoids. The 1H-NMR method does not require the use of reference compounds, and it requires only a short time for analysis. However, complex extracts in 1H-NMR may have a lot of signals, and quantitation with this method is often hampered by peak overlap, with 2D NMR providing a solution to this obstacle. The most important advantage of the COSY NMR quantitation method was the determination of the legality of cannabis plants, extracts, and edible oils based on their THC/THCA content, particularly in the cases of some samples for which the determination of THC/THCA content by 1H qNMR was not feasible

Quantitation of Oleuropein and Related Phenolics in Cured Spanish-Style Green, California-Style Black Ripe, and Greek-Style Natural Fermentation Olives.

Oleuropein, ligstroside, and related hydrolysis products are key contributors to olive bitterness, and several of these phenolics are implicated in the prevention of lifestyle age-related diseases. While table olive processing methods are designed to reduce oleuropein, the impact of processing on ligstroside and related hydrolysis products (e.g., oleacein, oleocanthal, hydroxytyrosol glucoside, ligstroside aglycone, and oleuropein aglycone) is relatively unknown. Herein, levels of these compounds were measured in Spanish-style green (SP), Californian-style black ripe (CA), and Greek-style natural fermentation (GK) olives using rapid ultrahigh-performance liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS). GK olives had the highest concentration of all compounds measured, with the exception of oleocanthal, which was highest in SP olives (0.081 mg kg-1 wet weight (w.wt)). CA olives had the lowest levels of most compounds measured, including ligstroside (0.115 mg kg-1 w.wt) and oleuropein (0.974 mg kg-1 w.wt). Hydroxytyrosol was the predominate compound in all three styles of commercial olives, with similar concentrations observed for GK and SP olives (134.329 and 133.685 mg kg-1 w.wt, respectively) and significantly lower concentrations observed for CA olives (19.981 mg kg-1 w.wt)