Polyphenolics total, determination

To this purpose, thirty-one different tomato cultivars were analyzed by spectrophotometric, chromatographic and mass spectrometric methods. Firstly, the content of flavonols (rutin and a mtin-pentoside), flavanones (naringenin and its chalcone), cinnamic acid derivatives (chlorogenic acid, caffeic acid, ferulic acid and a chlorogenic acid analogue), carotenoids (lycopene, P-carotene and lutein) and total polyphenols were determined. 

The content of total polyphenols was determined by colorimetric assay with Folin-Ciocalteu s phenol reagent, according to Singleton and Rossi 14). 

In the wine industry, FTIR has become a useful technique for rapid analysis of industrial-grade glycerol adulteration, polymeric mannose, organic acids, and varietal authenticity. Urbano Cuadrado et al. (2005) studied the applicability of spectroscopic techniques in the near- and mid-infrared frequencies to determine multiple wine parameters alcoholic degree, volumic mass, total acidity, total polyphenol index, glycerol, and total sulfur dioxide in a much more efficient approach than standard and reference methods in terms of time, reagent, and operation errors. 

Alonso AM, Guillen DA, Barroso CG, Puertas B and Gracia A. 2002. Determination of antioxidant activity of wine byproducts and its correlation with polyphenolic content. J Agric Food Chem 50 5832-5836. Al-Saikhan MS, Howard LR and Miller JC Jr. 1995. Antioxidant activity and total phenolics in different genotypes of potato (Solarium tuberosum L). J Food Sci 60 341-343, 7. 

Plant susceptibility to ozone as determined by visible injury may be very closely related to quantities of o-diphenols associated with the chloroplasts and specific requirements for activation of polyphenol oxidase enzymes. There is a significant correlation between ozone injury and concentrations of total phenols expressed as percent caffeic acid equivalents in peanut cultivars. This concept is not intended to underestimate the importance of membranes that separate phenols and enzymes. Perhaps future research will demonstrate that membranes of resistant alfalfa, green bean and other species differ both qualitatively and quantitatively from those of susceptible plants of these species. 

The total antioxidant activity of teas and tea polyphenols in aqueous phase oxidation reactions has been determined using an assay based on oxidation of 2,2,-azinobis-(3-ethyl benzothiazoline-sulfonate) (ABTS) by peroxyl radicals (114—117). Black and green tea extracts (2500 ppm) were found to be 8—12 times more effective antioxidants than a 1-mM solution of the water-soluble form of vitamin E, Trolox. The most potent antioxidants of the tea flavonoids were found to be epicatechin gallate and epigallocatechin gallate. A 1-mM solution of these flavanols were found respectively to be 4.9 and 4.8 times more potent than a 1-mM solution of Trolox in scavenging an ABT 5r+ radical cation. 

Dr. Joe Vinson of the University of Scranton believes that there is something to the chocolate effect, and he came to Belmont to tell us about his intriguing research. Vinson has determined the total polyphenol content of various chocolates and has also found a way of measuring how effective these mixtures are in preventing the oxidation of human ldl in a test tube. In other words, he has calculated a phenol antioxidant index, which takes into account both the quantity and the quality of these desirable substances. At the symposium Vinson reported that cocoa powder and dark chocolate are the best, followed by milk chocolate. Instant cocoa mixes trail the field. Then Vinson delivered the kicker chocolate has more, and better, polyphenols than fruits or vegetables and more than red wine. A forty-gram bar of dark chocolate has as many polyphenols as a cup of that widely promoted antioxidant cocktail we call tea. But there is still the matter of chocolate s fat content. Researchers tell us, though, that at least half of it is stearic acid, which does not raise blood cholesterol. 

For phenolics in fruit by-products such as apple seed, peel, cortex, and pomace, an HPLC method was also utilized. Apple waste is considered a potential source of specialty chemicals (58,62), and its quantitative polyphenol profile may be useful in apple cultivars for classification and identification. Chlorogenic acid and coumaroylquinic acids and phloridzin are known to be major phenolics in apple juice (53). However, in contrast to apple polyphenolics, HPLC with a 70% aqueous acetone extract of apple seeds showed that phloridzin alone accounts for ca. 75% of the total apple seed polyphenolics (62). Besides phloridzin, 13 other phenolics were identified by gradient HPLC/PDA on LiChrospher 100 RP-18 from apple seed (62). The HPLC technique was also able to provide polyphenol profiles in the peel and cortex of the apple to be used to characterize apple cultivars by multivariate statistical techniques (63). Phenolic compounds in the epidermis zone, parenchyma zone, core zone, and seeds of French cider apple varieties are also determined by HPLC (56). Three successive solvent extractions (hexane, methanol, aqueous acetone), binary HPLC gradient using (a) aqueous acetic acid, 2.5%, v/v, and (b) acetonitrile fol-... 

To construct and apply a biosensor based on capric acid/graphite powder modified with crude extract of jack fruit (Artocarpus integrifolia L.) as the source of polyphenol oxidase to determine total phenols in wastewaters by differential pulse voltammetry (DPV). 

Figure 8.2 Pomegranate juice polyphenols and antioxidant potency in comparison to other fruit juices. Total polyphenol concentration in the different juices was determined using quercetin as a standard. LDL (100 pg of protein/milliliter) was preincubated with increasing volume concentration (0-25 pL) of the juices. Then, 5 pmol/L of CuS04 was added, and the LDL was further incubated for 2 hours at 37°C. The extent of LDL oxidation was measured by the thiobar-bituric acid reactive substance (TBAR) assay, and the ICh, values (the concentration needed to get 50% inhibition) are given. Results are given as mean S.D. of three different experiments.

Polymeric fractions were obtained from wines, seed and skin extracts by fractionation on a Toyopearl HW-40 column as described by Souquet et al (4). Two aliquots of the fractions containing polymeric material were t en to dryness under vacuum. The first one was used to determine proanthocyanidin composition by thiolysis followed with HPLC analysis (17). The other one was dissolved in MeOH acidified with 2% HCl and used to estimate the concentration of total polymeric polyphenols and polymeric pigments by measuring the absorbance, respectively at 280 nm and 530 nm. Absorbance data were converted to equivalent epicatechin and equivalent malvidin-3-glucoside, respectively, using the extinction coefficients determined for each compounds under similar conditions. 

Fig. 10.5. Simplification of the determination of total polyphenols in virgin olive oil by use of a Fl-robot integrated approach. (Reproduced with permission of Elsevier Science.)...

There is good evidence that our bodies react to antioxidant compounds such as polyphenols as foreign and unwanted in high quantities, so the body actively metabolizes and excretes them, leaving behind only small amounts—about 3 percent—of the total amount ingested. Also, the concentration of polyphenols remaining in the body hours after a meal is tens to hundreds of times lower than the amount first tested in the lab to determine ORAC, making the lab measurement irrelevant in the human body. 

Total antioxidant activities in 13 typical Italian wines were determined with an average of 12.3 pM Trolox equivalents for red wines and 1.6 pM for white wines (Simonetti et al., 1997). The values correlated well with total phenols (r = 0.9902) and flavanols (r = 0.9270) and clearly demonstrated that red wine polyphenols are significant in vitro antioxidants. The antioxidant capacity of 16 red wines from several countries was tested using electron spin resonance (ESR) spectroscopy (Bums et al., 2000). The antioxidant activity ranged from 4.13 x 1021 to 9.29 x 1021 number of Fremy s radicals reduced by 1 L of wine. The antioxidant capacity was associated with the phenolic content of the wines, determined either by Folin-Ciocalteau or by summation of the individual phenolics determined by HPLC or by spectroscopy. Total phenolics as measured by the Folin-Ciocalteau method ranged from 6.47 to 18.6 mM GAE per liter. The anthocyanin levels ranged from 101.5 to 325.7 pM. There were substantial differences in the proportion of polymeric pigments in the wines. 

Clarification of the must in winemaking is made by treatments with silica gel, filtration, centrifugation, or by the use of enzymes. During filtration and centrifugation, oxidation of polyphenols may occur loss of protective colloids occurring with enzymes can promote precipitation of potassium bitartrate affecting the tartaric acid, pH and total acidity data. For determination of organic acids either in skins or in the... 

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