Fractioning of Proanthocyanidins of Uncaria tomentosa. Composition and Structure-Bioactivity Relationship

In a previous study, the detailed low-molecular weight polyphenolic profile of the different plant parts (leaves, stem, bark and wood) of Uncaria tomentosa was reported, the leaves being the plant part with the highest phenolic content and presenting the most heterogenous proanthocyanidin composition. Further, cytotoxicity of leaves extracts in two cancer cell lines was also found to be higher than in the remaining parts of the plant. In the present study, fractioning of U. tomentosa leaves polyphenolic extracts was performed using Diaion® HP-20 resin and a detailed characterization and quantification of fractions (n = 5) was achieved using advanced analytical techniques such as Ultra-Performance Liquid Chromatography coupled with Electrospray Ionization and Triple Quadrupole (TQD) Tandem Mass Spectrometry (UPLC/TQ-ESI-MS) and 13C-NMR. Oxygen Radical Absorbance Capacity (ORAC) and cytotoxicity on gastric adenocarcinoma AGS and colon adenocarcinoma SW20 cell lines were also determined in the different fractions. Results showed selective distribution of 32 non-flavonoid and flavonoid phenolics among the different fractions. ORAC varied between 3.2 and 11.8 µmol TE/mg in the different fractions, whereas IC50 of cytotoxicity on gastric adenocarcinoma AGS and colon adenocarcinoma SW20 cell lines best values were between 71.4 and 75.6 µg/mL. Fractions rich in proanthocyanidins also showed the highest bioactivity. In fact, significant positive correlation was found between total proanthocyanidins (TP) quantified by UPLC-DAD and ORAC (R 2 = 0.970), whereas significant negative correlation was found between TP and cytotoxicity towards AGS (R 2 = 0.820) and SW620 (R 2 = 0.843) adenocarcinoma cell lines. Among proanthocyanidins, propelargonidin dimers were of particular interest, showing significant correlation with cytotoxic selectivity on both gastric AGS (R 2 = 0.848) and colon SW620 (R 2 = 0.883) adenocarcinoma cell lines. These results show further evidence of the bioactivity of U. tomentosa proanthocyanidin extracts and their potential health effects.Agencia Española de Cooperación Internacional para el Desarrollo/[A/023397/09]/AECID/EspañaAgencia Española de Cooperación Internacional para el Desarrollo/[A/030037/10]/AECID/EspañaCosta Rica-USA Foundation/[]/CRUSA/Costa RicaConsejo Superior de Investigaciones Científicas/[CR0024]/CSIC/EspañaUniversidad de Costa Rica/[]/UCR/Costa RicaUCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de QuímicaUCR::Vicerrectoría de Docencia::Salud::Facultad de Medicina::Escuela de MedicinaUCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biologí

Comparative effects of A- and B-type proanthocyanidins in the prevention of urinary tract infection in mice

Resumen del póster presentado a la VI International Conference on Polyphenols and Health celebrada en Buenos Aires (Argentina) del 16 al 19 de octubre de 2013.Consumption of cranberry (Vaccinium macrocarpum) is widely recommended forprophylaxis against urinary tract infections (UTI) in women. Among cranberry components, A-type proanthocyanins would be implicated in these preventive effects against UTI. However, proanthocyanidins are poorly absorbed in the small intestine, but subjected to extensive biotransformation in the colon, although studies are almost restricted to B-type proanthocyanidins. Therefore, the hypothesis of this study is that urinary metabolome from of A-type and B-type proanthocyanidins-mainly derived from their colonic catabolism-differ,and only metabolites from the A-type procyanidins have protective effects against UTI. To test this hypothesis, JAXc3H/OuJ female mice previously fed with specific diet (control, 1% cranberry extract and 1% grape seed extract) for 2 weeks, were inoculated with the uropathogenic E. coli (ATCC 53503™) to provoke infection, and maintained 2 weeks more before being sacrificed. Urine samples were collected at different times and subjected to E.coli counting, leukocytary esterase and nitrites analyses, and mieloperoxidase task. Samples of kidney and bladder tissues were also collected for E. coli counting and histopathologic analysis. Additionally, the capacity of the urine samples to inhibit bacterial adherence was tested in the T24 bladder cell line (ATCC HTB4 ™).Peer reviewe

Effect of the modifier (Graciano vs. Cabernet sauvignon) on blends of Tempranillo wine during ageing in the bottle. II. Colour and overall appreciation

The effect of Graciano (GRA) (Spanish valuable variety of limited production) vs. Cabernet Sauvignon (CS) (world-wide known French variety) on the colour of wines from Tempranillo (TEM-base wine) (largely cultivated Spanish variety), was studied in wine blends prepared with 25% and 10% (v/v) of each modifier after 4, 6, 9, 16.5 and 23 months of ageing in the bottle. Although possessing pH values very similar to the base wine, the blends of TEM with GRA or CS wines showed chromatic changes (higher a*, C*, CI, %red and %dA, and lower h, %yellow, tint and L*) that were perceptible by the human eye, even when using as little as 10% of modifier wine. However, no differences in colour parameters were found between the TEM-GRA (90:10) and the TEM-CS (90:10) blends, and between the TEM-GRA (75:25) and the TEM-CS (75:25) ones, this being consistent with the results relating to the temporal evolution of anthocyanins and flavanols (Part I). Moreover, the results of the principal component analysis indicated that the degree of interrelation existing between the colour parameters and the phenolic components during ageing in the bottle was similar for the TEM-GRA and TEM-CS blends. From a practical point of view, the ANOVA analysis also demonstrated that for any of the modifier wines used, certain colour parameters and phenolic components allowed differentiation between the base wine and the 75:25 blends, whereas others allowed differentiation between the base wine and both the 90:10 and 75:25 blends, during the ageing period studied. Some similarities between GRA and CS as modifier wines of TEM blends were also found in terms of their organoleptic characteristics, also demonstrating that both varieties could render blended wines with better overall quality attributes than the base wine after 23 months of ageing in the bottle. © 2006 Swiss Society of Food Science and Technology.Spanish Comisión Interministerial de Ciencia y Tecnología (CICYT) (Project AGL2003-07394-C02-02Peer Reviewe

Time course of the colour of young red wines from Vitis vinifera L. during ageing in bottle

The colour characteristics of red wines from Vitis vinifera L. cv Tempranillo, Graciano and Cabernet Sauvignon (vintage 2000) from Navarra (Spain), was studied during 26 months of ageing in bottle through the evaluation of the wine visible spectrum and of several colorimetric indices (colour intensity, %red, %yellow, %blue, %dA and tint) and CIELAB variables (L*, C*, h, a* and b*). During ageing in bottle, the spectrum of Tempranillo wine (pH 4.3) mainly changed in the absorbance range between 420 and 500 nm, whereas Graciano (pH 3.5) and Cabernet Sauvignon (pH 3.6) wines registered a decrease in absorbance in the interval between 500 and 560 nm. The time course of the different wine colour parameters was found to fit either a second-order polynomium or a linear model, depending on the grape variety employed. CIELAB variables could be described in terms of their colorimetric index counterparts, showing b* and h relative greater errors. Although the wine total chromatic changes in CIELAB units registered after 26 months of ageing in bottle indicated changes perceivable by the human eye (ΔE* ≥ 2.7) for the three varieties studied, Graciano and Cabernet Sauvignon wines showed a more balanced colour evolution than Tempranillo wine.The authors also thank the Agencia Española de Cooperación International (AECI) for a MUTIS predoctoral scholarship to M.M. and the Spanish Comisión Interministerial de Ciencia y Tecnología (CICYT) (Project AGL2003-07394-C02-02) for funding.Peer reviewe

Statistical interpretation of the color parameters of red wines in function of their phenolic composition during aging in bottle

The relationships between the color parameters (colorimetric indexes and CIELAB variables) and the phenolic components (anthocyanins, pyranoanthocyanins, hydroxybenzoic and hydroxycinnamic acids, flavanols and flavonols) of young red wines from Vitis vinifera L. cv. Tempranillo, Graciano and Cabernet Sauvignon (vintage 2000, Navarra, Spain), have been investigated during 26 months of wine aging in bottle (a period embracing their commercial life), through the application of different statistical analysis (principal component, correlation and polynomial regression). The results of the principal component analysis first indicated that for each variety a high degree of interrelation existed between the color parameters and the anthocyanin pigments. Moreover, it was found that for each variety the color parameters were correlated with the anthocyanins during aging in bottle. Finally, by the application of polynomial regression analysis, both anthocyanins (simple glucosides and acetyl-glucosides) and pyranoanthocyanins (anthocyanin-pyruvic acid adducts) were selected as the variables that best described the different color parameters during aging in bottle. However, differences were found between varieties in relation to the type of anthocyanin pigment selected for describing each color parameter, finally indicating the importance of the grape variety factor in the definition of the wine chromatic characteristics.The authors thank the Agencia Española de Cooperación International (AECI) for a MUTIS predoctoral scholarship to M.M. and the Spanish Comisión Interministerial de Ciencia y Tecnología (CICYT) (Project ALI2003-07394-C02-02) for funding.Peer reviewe

Comprehensive assessment of the quality of commercial cranberry products. Phenolic characterization and in vitro bioactivity

Cranberry (Vaccinium macrocarpon) products have been widely recommended in traditional American medicine for the treatment of urinary tract infection (UTI). A total of 19 different commercial cranberry products from American and European markets have been analyzed by different global phenolic methods and by UPLC-DAD-ESI-TQ MS. In addition, in vitro antioxidant capacity and uropathogenic bacterial antiadhesion activity tests have been performed. Results revealed that products found in the market widely differed in their phenolic content and distribution, including products completely devoid of flavan-3-ols to highly purified ones, either in A-type proanthocyanidins (PACs) or in anthocyanins. The product presentation form and polyphenolic profile widely affected the antiadhesion activity, ranging from a negative (nulel) effect to a MIC = 0.5 mg/mL for cranberry powders and a MIC=112 mg/mL for gel capsule samples. Only 4 of 19 products would provide the recommended dose of intake of 36 mg total PACs/day. Of most importance was the fact that this dose would actually provide as low as 0.00 and up to 205 μg/g of procyanidin A2, indicating the lack of product standardization and incongruence between global and individual compound analysis. © 2012 American Chemical Society.This work has been funded by the Spanish Ministry for Science and Innovation (AGL2010-17499) and the Spanish Research Council (CSIC) (Programa Intramural Especial 201070I028). F.S.-P. is the recipient of a contract from the JAE-Doc Program (CSIC).Peer Reviewe

Almond (Prunus dulcis (Mill.) D.A. Webb) polyphenols: From chemical characterization to targeted analysis of phenolic metabolites in humans

10 páginas, 3 figuras, 3 tablas.-- El pdf del artículo es la versión de autor.In this paper, a survey of our studies on almond polyphenols including their chemical characterization and further bioavailability in humans is reported. Combination of analytical techniques (LC-DAD/fluorescence, LC/ESI-MS and MALDI-TOF-MS) allowed us, for the first time, the identification of A- and B-type procyanidin, propelargonidin and prodelphinidin polymers in almond skins. Glucuronide, O-methyl glucuronide, sulfate and O-methyl sulfate derivatives of (epi)catechin, as well as the glucuronide conjugates of naringenin and isorhamnetin, and sulfate conjugates of isorhamnetin, together with conjugates of hydroxyphenylvalerolactones were detected in plasma and urine samples after the intake of almond skin polyphenols. In addition, numerous microbial-derived metabolites, including hydroxyphenylpropionic, hydroxyphenylacetic, hydroxycinnamic, hydroxybenzoic and hydroxyhippuric acids were also identified. Depending of the type of metabolite, maximum urinary excretion was attained at different time in comparison to the control group in the course of the 24-h period of urine excretion, allowing us to establish the onset of microbial metabolism.This work has received financial support from the CONSOLIDER INGENIO 2010 Programme: FUN-C-FOOD CSD2007- 063 from the Spanish Ministry of Science and Innovation (MICINN). I.G. was the recipient of a fellowship from the I3P Program funding by the European Social Fund. M.U.-S. thanks the FPI fellowship from MICINN. M.M. thanks the ‘‘Ramón y Cajal” post-doctoral program. R.L.L. thanks the ‘‘Fondo de Investigación Sanitaria” postdoctoral program (F.I.S. CD06/00161) from MICINN.Peer reviewe

Quality assessment of commercial dietary antioxidant products from Vitis vinifera L. grape seeds

Phenolic preparations from Vitis vinifera L. grape seeds are products commonly used in the formulation of dietary antioxidant supplements. In this article, we used a methodology (the oxygen radical absorbance capacity, ORAC) to evaluate the in vitro antioxidant capacity of commercial dietary grape seed products and studied the relationship of the antioxidant capacity with the phenolic composition of these products. The ORAC value of the different brands of commercial products studied (n = 16) varied from 2.71 to 26.4 μmol Trolox equivalents/mg (≈10-fold difference). For four of these products, the batch-to-batch ORAC variation, expressed as a coefficient of variation of the mean, was 10.5% (n = 6), 13.1% (n = 3), 19.4% (n = 4), and 7.8% (n = 4). Analysis of monomeric and oligomeric flavan-3-ols by liquid chromatography-diode array detection (LC-DAD)/electrospray-mass spectrometry and procyanidins by thiolysis-LC-DAD also revealed large differences among the commercial grape seed products. Moreover, the ORAC value could be fitted to a regression model using variables from contents of individual phenolic compounds and procyanidins. The product-to-product and batch-to-batch variation in ORAC values and flavan-3-ol composition found among the commercial products studied demonstrated that they are poorly standardized, resulting in inconsistent composition and biological activity.Peer reviewe

Functional ingredients from Vitis vinifera L. commercialised in Europe

Trabajo presentado al XXIX Congreso Mundial de la Viña y el Vino, celebrado en España en junio de 2006.Peer reviewe