Several natural drugs (termed prodrugs) when administered orally undergo transformation by intestinal bacteria, producing metabolites, which may be more active than the parent compound. Mangiferin (I) is reported to have very low bioavailability in the upper gastrointestinal tract and reaches the large intestine, where it may be metabolized by the indigenous bacteria. Therefore, the aim of this study was to conduct pilot anaerobic fermentation studies with fecal inocula from human volunteers ( n = 3) to identify possible metabolic end products of mangiferin by the gastrointestinal metabolome. The major metabolite identified was deglycosylated mangiferin, namely, norathyriol (II) with an increase in homomangiferin (III) which was a minor contaminant of I. Mangiferin metabolites were identified and quantitated in the fermentation broths by high performance liquid chromatography (HPLC)–diode array detection–electrospray ionization-mass spectrometry, and structures confirmed unequivocally by nuclear magnetic resonance, after purification by semipreparative HPLC. Cell culture assays with 2 human cancer cell lines Caco-2 (colon cancer) and A240286S (non-small lung adenocarcinoma) showed that while the substrate mangiferin (I) and homomangiferin (III), a minor metabolite, are non-cytotoxic (half-maximal inhibitory concentration [IC 50 ] ≥ 100 µM), the major metabolite norathyriol (II) is cytotoxic against Caco-2 cells (IC 50 = 51.0 µM), whereas it is cytostatic against A240286S cells with a similar IC 50 (51.1 µM).

Pits of representative green olive drupes from Italy and Greece were evaluated for their content of phenolic compounds. The major purified polyphenols quantitated in mg/kg in the Italian and Greek pit varieties, respectively, were tyrosol (157 and 523), hydroxytyrosol (3 386 and 1723), vanillin (151 and 83), vanillic acid (58 and 101), phloretic acid (85 and 0), (+)-pinoresinol (954 and 877), (+)-1-acetoxypinoresinol (114 and 105), dehydrodiconiferyl alcohol (340 and 386) and the corresponding aldehyde (398 and 553), the erythro and threo isomers of guaiacylglycerol- β-coniferol ether (78 and 147) and (93 and 273) and their corresponding aldehydes (238 and 349) and (232 and 537), and finally nüzhenide (186 and 364). The data indicate that the waste products of olive mills should be a rich source of a variety of polyphenolic compounds. Detailed 1 H- and 13 C-nuclear magnetic resonance data are presented, representing the most comprehensive and unambiguous results available to date for the compounds studied.

High performance liquid chromatography coupled with negative electrospray ionization (HPLC–ESI) along with fragmentation patterns generated by nano-electrospray ionization (nano-ESI-MS–MS) and NMR techniques were utilized for the identification of phenolic compounds in Argan fruits. A total of 15.4 g/kg was determined represented by catechins (39%), flavonoids (28%), procyanidins (26%), free phenolic acids (6%) and phenolic acid glycosides (1%). Twenty-one phenolic compounds were identified for the first time in Argan fruits namely III. epicatechin-(4β → 8)-catechin dimer (procyanidin B1), IV. p-coumaric acid glycoside, VI. epicatechin-(4β → 8)-epicatechin dimer (procyanidin B2), VIII. caffeic acid glycoside, XIX. epicatechin-(4β → 8)-epicatechin-(4β → 8)-epicatechin trimer (procyanidin C1), X. p-hydroxybenzaldehyde XI. ferulic acid glycoside, XII. vanillic acid, XIII. sinapic acid glycoside, XVI. p-coumaric acid, XVII. ferulic acid, XVIII. sinapic acid, XIX. rutin pentoside, XX. quercetin glycopentoside, XXI. 4,4′-dihydroxy-3,3′-imino-di-benzoic acid, XXV. quercetin-3-O-rhamnogalactoside, XXVII. quercetin glycohydroxybenzoate, XXVIII. quercetin glycocaffeate, XXIX. quercetin glycosinapate, XXX. quercetin glycoferulate and XXXI. quercetin glycocoumarate.

AIMTo analyze phospholipid profiles in intrahepatic bile from patients with primary sclerosing cholangitis (PSC) and secondary sclerosing cholangitis (SSC).METHODSIntrahepatic bile specimens collected via endoscopic retrograde cholangiography from 41 patients were analyzed. Fourteen of these patients were diagnosed with PSC, 10 with SSC, 11 with choledocholithiasis or no identifiable biliary disease, and 6 with cholangiocellular carcinoma (CCC). Bile acid, cholesterol, protein, and bilirubin contents as well as pancreas lipase activity in bile were determined by biochemical methods. Phosphatidylcholine (PC) and lysophosphatidylcholine (LPC) species were quantified using nano-electrospray ionization tandem mass spectrometry.RESULTSBile from all the examined patient groups showed a remarkably similar PC and LPC species composition, with only minor statistical differences. Total biliary PC concentrations were highest in controls (8030 ± 1843 μmol/L) and lowest in patients with CCC (1969 ± 981 μmol/L) (P = 0.005, controls vs SSC and CCC, respectively, P < 0.05). LPC contents in bile were overall low (4.2% ± 1.8%). Biliary LPC/PC ratios and ratios of biliary PC to bilirubin, PC to cholesterol, PC to protein, and PC to bile acids showed no intergroup differences.CONCLUSIONPC and LPC profiles being similar in patients with or without sclerosing cholangitis, these phospholipids are likely not of major pathogenetic importance in this disease group.

Mirabelle plums represent a famous fruit from the Lorraine region, however little is known about their phytochemical composition. The oil of the fruit contained predominantly oleic acid (59%) and linoleic acid (29%). The total content of phenolic antioxidants in the whole fruits was 5.338g/kg with 456mg/kg (9%), 701mg/kg (13%) and 4159mg/kg (78%) detected in the peels, flesh and pits respectively. The peels contained solely 3,4-dihydroxybenzoic acid (270mg/kg) and rutin (186mg/kg), the flesh exclusively echinoids (723mg/kg), whereas the pits contained a rich variety of phenolic compounds (4.2g/kg) dominated by amygdalin (3.8g/kg), but with significant contributions from vanillin (102mg/kg), guajacyl-glycerin-coniferyl aldehyde isomers (87mg/kg), dehydro-diconiferyl aldehyde (52mg/kg), and vanillin diglucoside (48mg/kg). Of the major phenolic compounds tested across a range of in vitro assays, rutin was the superior antioxidant.

Longan (Dimocarpus longan Lour, syn. Euphoria longan Lam.) represents an important fruit in Northern Thailand and has significant economic impact. The fruit is either consumed fresh or as commercially prepared dried and canned products. The canning industry in Thailand produces considerable quantities of waste products, in particular Longan seeds. Because these seeds may be an exploitable source of natural phenolic antioxidants, it was of interest to identify, purify and quantitate the major potential antioxidant phenolics contained therein. The polyphenolic fraction from ground Longan seeds was obtained by extraction with methanol after delipidation with hexane. The hexane extract contained predominantly long-chain fatty acids with major contributions from palmitic (35%) and oleic (28%) acids. The polyphenolic fraction (80.90 g/kg dry weight) was dominated by ellagic acid (25.84 g/kg) and the known ellagitannins corilagin (13.31 g/kg), chebulagic acid (13.06 g/kg), ellagic acid 4-O-α-l-arabinofuranoside (9.93 g/kg), isomallotinic acid (8.56 g/kg) and geraniin (5.79 g/kg). Structure elucidation was performed with mass spectrometry and complete assignment of 1H and 13C NMR signals. The methanol extracts exhibited strong antioxidant capacities with an IC50 of 154 μg/ml for reactive oxygen species attack on salicylic acid and 78 μg/ml for inhibition of xanthine oxidase in the hypoxanthine/xanthine oxidase assay. The extracts were less effective in the 2-deoxyguanosine assay (IC50 = 2.46 mg/ml), indicating that gallates along with ellagic acid and its congeners exert their potential antioxidant effects predominantly by precipitation of proteins such as xanthine oxidase. This was confirmed for the pure compounds gallic acid, methyl gallate, ellagic acid and corilagin.

Thirty-four polyphenolic substances in methanol extracts of the fruits of Terminalia bellerica, Terminalia chebula and Terminalia horrida, three plants used in Egyptian folk medicine, were initially identified by HPLC–ESI-MS and quantitated by analytical HPLC after column chromatography on Sephadex LH-20. After purification by semi-preparative HPLC the compounds were identified by their mass and fragmentation patterns using ESI-MS–MS. For several compounds detailed 1H/13C NMR analysis at 600 MHz was performed. Two polyphenolics, namely 4-O-(4″-O-galloyl-α-l-rhamnopyranosyl)ellagic acid and 4-O-(3″,4″-di-O-galloyl-α-l-rhamnopyranosyl)ellagic acid were identified by NMR. Antioxidant capacities of the raw fruit extracts and the major isolated substances were determined using the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), oxygen radical absorbance capacity (ORAC) and ferric reducing ability of plasma (FRAP) in vitro assays and indicated that chebulic ellagitannins have high activity which may correlate with high potential as cancer chemopreventive agents. Therefore, further studies (metabolism, bioavailability and toxicity) of the polyphenolics in Terminalia species using preclinical models and in vivo human intervention trials are warranted.

Background: Phospholipids are essential for the normal function of the intestinal mucus barrier. The objective of this study was to systematically investigate phospholipids in the intestinal mucus of humans Suffering from inflammatory bowel diseases, where it barrier defect is strongly supposed to be pathogenetic.Methods: Optimal mucus recovery was first validated in healthy mice and the method was then transferred to the endoscopic acquisition of ileal and colonic mucus from 21 patients with ulcerative colitis (UC), 10 patients with Crohn's disease (CD), and 29 healthy controls. Nano-electrospray ionization tandem mass spectrometry (ESI-MS/MS) was used to determine phosphatidylcholine (PC), lysophosphatidylcholine (LPC), and sphingomyelin (SM) in lipid extracts of mucus specimens.Results: Human and rodent mucus contained very similar phospholipid species. In the ileal and colonic mucus from patients suffering from UC, the concentration of PC was highly significantly lower (607 +/- 147 pmol/100 mu g protein and 745 +/- 148 pmol/100 mu g protein) compared to that of patients with CD (3223 +/- 1519 pmol/100 mu g protein and 2450 +/- 431 pmol/100 mu g protein) and to controls (3870 +/- 760 pmol/100 mu g protein and 2790 +/- 354 pmol/100 mu g protein), overall, P = 0.0002 for ileal specimens and P < 0.0001 tor colonic specimens. Independent of disease activity, patients suffering from UC showed an increased Saturation grade of PC fatty acid residues and it higher LPC-to-PC ratio.Conclusions: The intestinal mucus barrier of patients with UC is significantly altered concerning its phospholipid concentration and species composition. These alterations may be very important for the pathogenesis of this disease and underline new therapeutic strategies.

Silybin or silymarin extract has been used to treat liver diseases, and has now been entered into clinical trials for cancer treatment. Here, we compared antioxidant and anticancer activities between silybin and its oxidized form 2,3-dehydrosilybin (DHS). With IC50 at three-fold lower concentrations than silybin, DHS inhibited reactive oxygen species generation in glucose-glucose oxidase system and HcpG2 cells. Compared with silybin, DHS elicited greater protection against H2O2-induced HepG2 cell death and galactosamine-induced liver injury in vivo. It is known that oxidants induce releases of metalloproteinases (MMP)-2,-9 which are responsible for invasive and metastasis potentials of transformed cells. DHS at 10 mu M markedly inhibited MMP-2,-9 releases as well as invasiveness, while silybin at 90 mu M had marginal effects. DHS but not silybin at 30 mu M induced apoptosis and loss of mitochondrial membrane potentials. LD50 of DHS was five-fold lower than that of silybin. Our data suggest that DHS may be more useful therapeutically than silybin. (C) 2007 Elsevier B.V. All rights reserved.

The contents of secondary plant substances in solvent extracts of various byproducts (barks, kernels, peels, and old and young leaves) in a range of Brazilian mango cultivars were identified and quantitated. The results show that the profiles of secondary plant substances such as xanthone C-glycosides, gallotannins, and benzophenones in different byproducts vary greatly but are fairly consistent across cultivars. The free radical scavenging activity of the solvent extracts was evaluated using a high-performance liquid chromatography-based hypoxanthine/xanthine oxidase assay and revealed dose-dependent antioxidant capacity in all extracts. Four (mangiferin, penta- O-galloyl-glucoside gallic acid, and methyl gallate) of the major phenolic compounds detected were also evaluated in additional in vitro bioassay systems such as oxygen radical absorbance capacity, 2,2-diphenyl-1-picrylhydrazyl, and ferric reducing ability of plasma. Mangiferin in particular, detected at high concentrations in young leaves (Coite = 172 g/kg), in bark (Momika = 107 g/kg), and in old leaves (Itamaraka = 94 g/kg), shows an exceptionally strong antioxidant capacity.

Very few relevant methods have been described for the detection and quantitation of phenolic compounds in faecal matrix. Extraction with conventional organic solvents such as chloroform/methanol (2:1, Folch reagent), methanol and ethanol (72%) showed high extraction efficiency for lipids and also gave good recovery of the major phenolic compounds present in the matrix. However, in comparison with a newly developed phosphate buffer method, the yield of minor phenolics was negligible when detected by these conventional methods. Conventional methods also lead to contamination of the ion source of the mass spectrometer and rapid deterioration of column performance mostly due to the high concentration of lipids. However, if the faecal matrix is initially extracted with phosphate buffer, and the extract acidified and re-extracted with diethyl ether, the range and yield of phenolic compounds are enhanced and the problem of lipid contamination is substantially alleviated. Following pilot studies and optimisation of the procedure, individual phenolic compounds (n = 29) were identified by nano-electrospray ionisation mass spectrometry (nano-ESI-MS), nano-ESI-tandem mass spectrometry (MS/MS) and gas chromatography/mass spectrometry (GC/EI-MS) and quantitated (n = 27) by GC/MS in subsets (n = 5) of faecal samples, collected during the European Agency for Cancer Prevention calcium/fibre intervention study from four European countries (Italy, Germany, Spain and Denmark). A range of phenolic compounds (mainly acids) was detected, dominated by phenylacetic, benzoic, phenylpropionic and m-hydroxyphenylpropionic acids, representing on average 9.91 (93%), 8.25 (92%), 9.45 (95%) and 11.05 (98%) mM in the Italian, German, Spanish and Danish samples, respectively. The new method should enable large epidemiologic, case-control and intervention studies on the relevance of phenolic antioxidants in the aetiology of colorectal cancer to be conducted in the future.