tannins and vanillin

Phenolic substances occur primarily in fruits and vegetables and in the seeds of certain pigmented cultivars of sorghum, millets, and legumes. One of the major difficulties encountered in polyphenol research is the lack of a standard quantitative method for the analysis of phenolics that would be suitable for a wide range of seeds, forage crops, and food products and under a variety of experimental conditions. Some methods measure "total phenol", which may not be a true index of the nutritional quality of foods and thus does not distinguish polyphenols of nutritional concern from other low-molecular-weight phenols that also occur naturally in these products. Tannic acid (a hydrolyzable gallotannin) is commonly used as a "reference standard", but this may be a questionable practice since its biological properties differ from those of tannins of flavonoid origin. Polyphenols of cereals and legumes are predominantly of the latter type. Also, commercially available tannic acid has been shown to be a mixture of four phenolic compounds, the relative proportions of which vary with the samples. Thus, the choice of a suitable standard for tannin analysis is also important. The quantitative extraction of the condensed tannins from plant tissue is always difficult, since it may be complexed to a carbohydrate or protein matrix which could be quite insoluble due to a high degree of polymerization. The literature on tannin methodology is diverse and at times conflicting. Currently available methods for tannin analysis range from simple colorimetric, UV spectrophotometric, chromatographic, and enzymic to more sophisticated and expensive nuclear magnetic resonance (NMR) techniques. None of these methods of analyses is completely satisfactory nor can it be applied to different food products with the same degree of success. This review covers physical and chemical methods for tannin analysis of different food products, the problems in analysis and interpretation of data, and future research needs in this area.

Topics: Benzaldehydes; Chemical Phenomena; Chemical Precipitation; Chemistry; Food Analysis; Oxidation-Reduction; Phenols; Spectrophotometry, Ultraviolet; Tannins

Sorghum (Sorghum bicolor (L.) Moench) contains a wide variety of phenolic compounds and their levels and types depend on the genotype. Sorghums with a pigmented testa contain condensed tannins, which are concentrated in the pigmented testa. In this chapter, three methods of tannin analysis are described: (1) Chlorox bleach test; (2) modified vanillin-HCl assay; and (3) normal-phase high-performance liquid chromatography (NP-HPLC) with fluorescence detection. The Chlorox bleach test is a rapid and inexpensive method that identifies non-tannin and tannin sorghums. The modified vanillin-HCl assay is a colorimetric assay that provides relative tannin values among sorghum samples. The normal-phase HPLC with fluorescence detection separates and quantifies condensed tannins according to their degree of polymerization and confirms the presence or absence of tannins.

Topics: Benzaldehydes; Chromatography, High Pressure Liquid; Edible Grain; Fluorescence; Phenols; Sorghum; Tannins

The major areas of the world where viticulture is practiced enjoy temperate or cool temperature climates. When wine grapes are grown in tropical regions, edaphoclimatic factors result in distinct grape quality attributes, and production techniques also require significant adjustment. The objective of this study was to characterize the chemical compositions, in particular of phenolic compounds, of Syrah grapes grown in two location in northeast Brazil - these are also at widely different altitudes. A range of methods of phenolic extraction were used, along with classical chemical analyses including for organic acids, sugars, monomeric anthocyanins, flavonols, stilbene, condensed tannins and some of the monomeric and small oligomeric procyanidins. The regions and their diverse environments had a larger influence than harvest year. The grapes at higher altitude (Bahia, 1.100 m asl (metres above sea level) were characterized by higher levels of malic acid, anthocyanins and condensed tannins in the skins. The low-altitude grapes (Pernambuco, 350 m asl (metres above sea level) had higher levels of glucose, fructose, 3-O-acetylglucoside anthocyanins and condensed tannins in the seeds. Fruit composition was highly influenced by the region. In the low-altitude region, the grapes were characterized by higher tartaric and citric acid in the must, also of flavonols in skins and of tannins in the seeds. Meanwhile, the fruit from the high altitude, contained higher levels of malic and succinic acid in the must, and of anthocyanins and condensed tannins in the skins.

Topics: Altitude; Anthocyanins; Antioxidants; Benzaldehydes; Biflavonoids; Brazil; Catechin; Chromatography, High Pressure Liquid; Color; Farms; Flavonols; Fructose; Fruit; Glucose; Molecular Weight; Phenols; Principal Component Analysis; Proanthocyanidins; Seeds; Stilbenes; Tannins; Tropical Climate; Vitis; Wine

An endo-β-1,4-xylanase (X22) was purified from crude extract of Emericella nidulans when cultivated on submerged fermentation using sugarcane bagasse as the carbon source. The purified protein was identified by mass spectrometry and was most active at pH and temperature intervals of 5.0-6.5 and 50-60°C, respectively. The enzyme showed half-lives of 40, 10 and 7 min at 28, 50 and 55°C, respectively, and pH 5.0. Apparent Km and Vmax values on soluble oat spelt xylan were 3.39 mg/mL and 230.8 IU/mg, respectively, while Kcat and Kcat/Km were 84.6 s(-1) and 25.0 s(-1) mg(-1) mL. Incubation with phenolic compounds showed that tannic acid and cinnamic acid had an inhibitory effect on X22 but no time-dependent deactivation. On the other hand, ferulic acid, 4-hydroxybenzoic acid, vanillin and p-coumaric acid did not show any inhibitory effect on X22 activity, although they changed X22 apparent kinetic parameters. Ethanol remarkably increased enzyme thermostability and apparent Vmax and Kcat values, even though the affinity and catalytic efficiency for xylan were lowered.

Topics: Benzaldehydes; Cellulose; Cinnamates; Coumaric Acids; Emericella; Endo-1,4-beta Xylanases; Enzyme Stability; Ethanol; Fermentation; Hydrogen-Ion Concentration; Kinetics; Lignin; Parabens; Propionates; Saccharum; Substrate Specificity; Tannins

The HCI-vanillin assay is a well-accepted method for determining tannin content in sorghum but is limited to small sample sets due to the time-consuming nature of the method. The objective was to develop an accurate and repeatable high-throughput 96-well plate assay for breeders to screen large sample sets of sorghum for tannin content. Validation of the high-throughput assay was tested on 25 sorghums suspected to contain tannin.. Approximately 30 measurements per day were completed using the conventional assay compared to 224 measurements using the 96-well platform. The correlation between the two tannin assays was 0.98. The coefficient of variation (CV) was 3.54% and 3.21% for the 96-well and conventional method, respectively. The 96-well assay exhibited good repeatability, with the inter-plate CV between 2.77% and 4.85%.. The high-throughput 96-well HCI-vanillin assay exhibited an eightfold increase in the number of measurements completed and was as accurate as the conventional HCI-vanillin assay.

Topics: Benzaldehydes; Edible Grain; High-Throughput Screening Assays; Humans; Reproducibility of Results; Seeds; Sorghum; Tannins

Phenolics derived from lignin and other plant components can pose significant inhibition on enzymatic conversion of cellulosic biomass materials to useful chemicals. Understanding the mechanism of such inhibition is of importance for the development of viable biomass conversion technologies. In native plant cell wall, most of the phenolics and derivatives are found in polymeric lignin. When biomass feedstocks are pretreated (prior to enzymatic hydrolysis), simple or oligomeric phenolics and derivatives are often generated from lignin modification/degradation, which can inhibit biomass-converting enzymes. To further understand how such phenolic substances may affect cellulase reaction, we carried out a comparative study on a series of simple and oligomeric phenolics representing or mimicking the composition of lignin or its degradation products. Consistent to previous studies, we observed that oligomeric phenolics could exert more inhibition on enzymatic cellulolysis than simple phenolics. Oligomeric phenolics could inactivate cellulases by reversibly complexing them. Simple and oligomeric phenolics could also inhibit enzymatic cellulolysis by adsorbing onto cellulose. Individual cellulases showed different susceptibility toward these inhibitions. Polyethylene glycol and tannase could respectively bind and degrade the studied oligomeric phenolics, and by doing so mitigate the oligomeric phenolic's inhibition on cellulolysis.

Topics: Benzaldehydes; Biomass; Cellulases; Cellulose; Hydrolysis; Lignin; Phenols; Tannins

The tannin fractions isolated from hazelnuts, walnuts and almonds were characterised by colorimetric assays and by an SE-HPLC technique. The complexation of Cu(II) and Zn(II) was determined by the reaction with tetramethylmurexide, whereas for Fe(II), ferrozine was employed. The walnut tannins exhibited a significantly weaker reaction with the vanillin/HCl reagent than hazelnut and almond tannins, but the protein precipitation capacity of the walnut fraction was high. The SE-HPLC chromatogram of the tannin fraction from hazelnuts revealed the presence of oligomers with higher molecular weights compared to that of almonds. Copper ions were most effectively chelated by the constituents of the tannin fractions of hazelnuts, walnuts and almonds. At a 0.2 mg/assay addition level, the walnut tannins complexed almost 100% Cu(II). The Fe(II) complexation capacities of the tannin fractions of walnuts and hazelnuts were weaker in comparison to that of the almond tannin fraction, which at a 2.5 mg/assay addition level, bound Fe(II) by approximately 90%. The capacity to chelate Zn(II) was quite varied for the different nut tannin fractions: almond tannins bound as much as 84% Zn(II), whereas the value for walnut tannins was only 8.7%; and for hazelnut tannins, no Zn(II) chelation took place at the levels tested.

Topics: Benzaldehydes; Chelating Agents; Copper; Corylus; Iron; Juglans; Nuts; Prunus; Tannins; Zinc

Phenolic compounds were extracted from red lentil seeds using 80% (v/v) aqueous acetone. The crude extract was applied to a Sephadex LH-20 column. Fraction 1, consisting of sugars and low-molecular-weight phenolics, was eluted from the column by ethanol. Fraction 2, consisting of tannins, was obtained using acetone-water (1:1; v/v) as the mobile phase. Phenolic compounds present in the crude extract and its fractions demonstrated antioxidant and antiradical activities as revealed from studies using a beta-carotene-linoleate model system, the total antioxidant activity (TAA) method, the DPPH radical-scavenging activity assay, and a reducing power evaluation. Results of these assays showed the highest values when tannins (fraction 2) were tested. For instance, the TAA of the tannin fraction was 5.85 micromol Trolox eq./mg, whereas the crude extract and fraction 1 showed 0.68 and 0.33 micromol Trolox eq./mg, respectively. The content of total phenolics in fraction 2 was the highest (290 mg/g); the tannin content, determined using the vanillin method and expressed as absorbance units at 500 nm per 1 g, was 129. There were 24 compounds identified in the crude extract using an HPLC-ESI-MS method: quercetin diglycoside, catechin, digallate procyanidin, and p-hydroxybenzoic were the dominant phenolics in the extract.

Topics: Benzaldehydes; beta Carotene; Chemical Fractionation; Free Radical Scavengers; Lens Plant; Phenols; Plant Extracts; Tannins

The leaves of Houttuynia cordata have been traditionally used as medicinal foods in East Asia. However, there have been few data about the biological effects. The antioxidative effects of polyphenols in the leaves of H. cordata were investigated on protein fragmentation by copper-hydrogen peroxide in vitro. The total polyphenol content in the leaves of H. cordata was 1.14%. The condensed tannin content was 2.46% by vanillin assay and 0.54% by proanthrocyanidin assay. The polyphenols in the leaves of H. cordata inhibited bovine serum albumin fragmentation by copper-hydrogen peroxide. These results demonstrated that the leaves of H. cordata have antioxidative effects on biological damage such as protein fragmentation.

Topics: Antioxidants; Benzaldehydes; Copper; Drugs, Chinese Herbal; Flavonoids; Houttuynia; Hydrogen Peroxide; Peptide Fragments; Phenols; Plant Leaves; Plant Proteins; Polyphenols; Serum Albumin, Bovine; Tannins

The chemical composition of cooperage oak wood is highly variable, depending upon the tree species (Quercus robur L. versus Quercus petraea Liebl.), its geographic location, and the single-tree effect. In the process of cask-making, natural seasoning and toasting contribute strongly to the modification of the oak wood chemical composition and therefore influence wine cooperaging. HPLC and GC quantification of ellagitannins and volatile compounds such as whiskey-lactones, eugenol, and vanillin over a sample set of 61 pedunculate oaks and 72 sessile oaks originating from six different forests showed that natural drying leads to a decrease of the ellagitannins and total extractives content level and a quasi constant level of the volatile compounds. Toasting (medium type) drastically enhanced the loss of ellagitannins and the gain in volatile compounds. Statistical treatment showed that the species effect remained significant throughout the process of drying and toasting, but not the provenance. The poor correlation with ring width of extractives levels measured on fresh timber remained unchanged as did the single-tree effect, with high variability found for all chemical parameters. These results provide further evidence that cooperage oak selection should not be based solely on the wood grain or the provenance but rather on a species-provenance combination.

Topics: Benzaldehydes; Chromatography, Gas; Chromatography, High Pressure Liquid; Desiccation; Eugenol; Hydrolyzable Tannins; Lactones; Quercus; Species Specificity; Tannins; Volatilization; Wine; Wood

Topics: Benzaldehydes; Chromatography, Gas; Eye Diseases; Gas Chromatography-Mass Spectrometry; Glycyrrhiza; Medicine, Chinese Traditional; Phytotherapy; Plant Extracts; Plants, Medicinal; Tannins