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Zbigniew Pietrzkowski,1 Ruby Argumedo,1 Cynthia Shu,1 Boris Nemzer,2 Slawomir Wybraniec,3 Tania Reyes-Izquierdo1 1Applied BioClinical Inc., Irvine, CA, USA; 2FutureCeuticals Inc., Momence, IL, USA; 3Department of Analytical Chemistry, Institute C-1, Faculty of Chemical Engineering and Technology, Cracow University of Technology, Cracow, Poland Aim: We aimed to evaluate the effect of a betalain-rich red beet concentrate (BRC) on joint discomfort and joint function. Methods: Individuals with self-reported knee discomfort were randomized and blinded to treatment with either oral BRC (50 mg twice a day) or placebo. Symptoms of discomfort and joint function were assessed using the Western Ontario and McMaster Universities Arthritis Index (WOMAC) and the McGill Pain Questionnaire (MPQ) before treatment and after 5 and 10 days of treatment. The general level of energy was evaluated using a visual analog scale (VAS) at the same time points. Results: In all, 40 individuals were evaluated in this study, 20 in the BRC group and 20 in the placebo group. The mean participant age was 55 years (range: 45–65). All participants completed the study. Individuals receiving BRC had a 27% better MPQ score after 10 days than did individuals in the placebo group (P=0.003). Furthermore, knee function, as measured by WOMAC score, was 26% better in the BRC group than in the placebo group (P

Tania Reyes-Izquierdo,1 Boris Nemzer,2 Ruby Argumedo,1 Cynthia Shu,1 Zb Pietrzkowski1 1Applied BioClinical Inc., Irvine, CA, 2FutureCeuticals Inc, Momence, IL, USA Background: Dermaval™ is a composite formulation of various phytochemical-rich plant materials as quantified using high-performance liquid chromatography. This blend exerts inhibitory activity on human neutrophil elastase (HNE). An acute, crossover clinical study was performed to assess the effects of Dermaval on glucose-induced HNE activity in 20 healthy subjects. Methods: Participants served as their own controls during this 3-day trial. On day 1, all study participants were fasted and given only 300 mL of water. Blood was drawn before treatment and 60 and 120 minutes post treatment. On day 2, participants were fasted, treated with 75 g of glucose, and similarly tested. On day 3, participants consumed a 50 mg serving of Dermaval followed by 75 g of glucose 15 minutes later. HNE concentration and HNE total activity were determined using enzyme-linked immunosorbent assay. Results: Average values for HNE activity in the control group (day 1) did not change. Treatment with a single dose of glucose (day 2) increased blood HNE activity by 175% over baseline levels (P=0.005) during the first 60 minutes. Pretreatment with Dermaval on day 3 prevented the glucose-induced increase in HNE activity (P=0.005 at 60 minutes and P=0.03 at 120 minutes versus respective day 2 values). Interestingly, ingestion of 75 g of glucose resulted in the same blood glucose levels on days 2 and 3, indicating that ingestion of Dermaval did not affect glucose absorption. Conclusion: These results suggest that Dermaval acutely inhibits glucose-induced HNE activity. Further investigations are needed to elucidate the direct and/or indirect mechanism of this effect and to verify whether treatment with Dermaval without glucose may acutely affect activity of HNE as well. Keywords: crossover study, human neutrophil elastase, glucose

Abstract Background Measuring the effects of the acute intake of natural products on human biomarker concentrations, such as those related to oxidation and inflammation, can be an advantageous strategy for early clinical research on an ingredient or product. Methods 31 total healthy subjects were randomized in a double-blinded, placebo-controlled, acute pilot study with post-hoc subgroup analysis on 20 of the subjects. The study examined the effects of a single dose of a polyphenol-rich beverage (PRB), commercially marketed as "SoZo®", on serum anti-inflammatory and antioxidant markers. In addition, phytochemical analyses of PRB, and in vitro antioxidant capacity were also performed. Results At 1 hour post-intake, serum values for 8-iso-PGF2-alpha and advanced oxidation protein products decreased significantly by 40% and 39%, respectively. Additionally, there was a trend toward decreased C-reactive protein, and increased nitric oxide levels. Both placebo and PRB treatment resulted in statistically significant increases in hydroxyl radical antioxidant capacity (HORAC) compared to baseline; PRB showed a higher percent change (55-75% versus 23-74% in placebo group), but the two groups did not differ significantly from each other. Conclusions PRB produced statistically significant changes in several blood biomarkers related to antioxidant/anti-inflammatory effects. Future studies are justified to verify results and test for cumulative effects of repeated intakes of PRB. The study demonstrates the potential utility of acute biomarker measurements for evaluating antioxidant/anti-inflammatory effects of natural products.

Human peripheral blood mononuclear cells (PBMC) were used as a biological model to measure intracellular ATP (iA TP) reactive oxygen species (ROS), Oxygen Consumption Rate (OCR) and extracellular acidification rate (ECAR), in response to acute ex vivo treatments with HH2o. "HH2o ", a fermented, food-based material commercially marketed as "Mitochroma™" has been identified as a potential modulator of iATP, OCR and ROS in these cells. PBMC were exposed to HH2o (0.1, 0.01 and 0.001% w/v) and Carbonyl-cyanide p-trifluoro-methoxy-phenyl-hydrazone (FCCP) or Oligomycin, two known modulators of mitochondrial activity were used as controls. Results showed that treatment ofthe cells with HH20 resulted in increase of iATP up to 220% and reduction of iADP by up to 40% while iROS levels remained unchanged. At the same time, OCR level was increased up to 24% compared to initial baseline. These results suggest that HH2o acts similarly to uncoupling protein 2 (UCP2) when induced and that it may have potential use in developing health-promoting products. These results support the use of freshly isolated human PBMCs as an experimental ex vivo model to detect the effects of nutraceutical products on dynamic cell metabolites, such as oxygen consumption, ATP, ADP and ROS. [ABSTRACT FROM AUTHOR]

Sprouted grains are gaining popularity as functional food ingredients. This study aimed to evaluate the lipid and fatty acid composition of eight sprouted grains (millet, amaranth, quinoa, wheat, rye, barley, buckwheat, and oat). The method used was germination for up to 72 h at temperatures ranging from 19-23 °C. In general, the lipid content increased in the various grains sprouted, providing a rich source of polyunsaturated fatty acids. The % oil yield ranged from 1.17 ± 0.02% in sprouted rye to 5.71 ± 0.26% in sprouted amaranth. Germinated oat showed the greatest increase in fat content, 54.3%, compared to the control. Polyunsaturated fatty acids were more prevalent in whole grains (46.9-75.6%) than saturated fatty acids (10.1-25.9%) and increased with sprouting. The primary fatty acids detected in the grains, in order of abundance, were linoleic, oleic, palmitic, linolenic, and stearic acids. Millet sprouts contained the lowest total saturated fatty acids and the highest polyunsaturated fatty acids. Amaranth had the highest amount of saturated fatty acids, while buckwheat contained the lowest quantity of polyunsaturated fatty acids. The lowest omega-6/omega-3 ratio was 7 to 1 in sprouted rye and 8 to 1 in sprouted barley.

A whole-grain diet is associated with the prevention of metabolic syndromes, including obesity, diabetes, and cardiovascular diseases. Sprouting improves the nutritional profile and bioactive properties of grains, which are important for use as raw ingredients in the food industry. The aim of this review was to examine the lipid and fatty acid composition of germinated grains. The methods discussed include germination and analytical procedures for determining fat and fatty acid contents of grains. The effects of sprouting on the fat content and storage stability of grains were also assessed. Lipid levels ranged from 1.43% to 6.66% in the sprouted grains. The individual fatty acid content of grains changed depending on the germination conditions (17-37 °C, 1-9 days). Limited findings showed that sprouting grains at higher temperatures (20-25 °C) and longer times generated a healthy balance of omega-6 and omega-3 fatty acids, which is beneficial to humans. Future studies are needed to determine the optimum incubation and germination periods specific to each grain to improve the omega-6/omega-3 ratio. Free fatty acids were produced more slowly and levels of oxidation products were lower in sprouted grains than in the raw ingredients when stored for a year. Additional studies are required to investigate the oxidative stability and shelf life of sprouted grains.

Coffee cherry is a rich source of caffeine and chlorogenic acids. In this study we investigate the structural analysis of caffeine-enriched whole coffee cherry extracts, CEWCCE by using 1 H and 13 C NMR spectroscopy. The changes in 1 H chemical shift data in NMR spectra of CEWCCE compared to pure caffeine indicated the formation of complexes between caffeine and chlorogenic acids in aqueous solution. The effect of complexation on the peak position of caffeoylquinic acid and caffeine resonance with increasing addition of caffeine was investigated. 2D NOESY experiments show the presence of cross-peaks that are due to the proximity of chlorogenic acid and caffeine molecules in stable complexes in protic solvents. The quantification data of caffeine by 1 H qNMR was found to be in close agreement with the data obtained by HPLC analysis.

The aim of the study was to evaluate the possible correlation between the bioactivity and the phytochemical profile of four betalain-rich extracts from Portulaca grandiflora Hook. The HPLC-DAD-ESI-MS analysis indicated the presence of 19 betaxanthins and two betacyanins. The highest concentrations of betaxanthins (982 mg/100 g DE) and betacyanins (650 mg/100 g DE) were noticed in orange and purple flowers extracts, respectively. The HPLC-DAD-ESI-HRMS/MS analyses revealed the presence of a total of 71 compounds. Fifteen new betaxanthins and fifty other metabolites were identified for the first time. The antioxidant activity of the studied flower extracts increased in the sequence of yellow < orange < purple < red (0.066−0.176 mM TE/g DE). Betalains showed less effect on the antioxidant activity of extracts than other metabolites did. Extracts from yellow and orange flowers were more active against Gram-positive bacteria (MIC = 4−16 mg/L), whereas extracts from red and purple flowers were slightly more active against Gram-negative bacteria (MIC = 16−32 mg/L). All the extracts showed the same activity against yeasts (MIC = 32 mg/L). Betaxanthins were active against Gram-positive bacteria, whereas betacyanins were active against Gram-negative bacteria. Remaining metabolites also exhibited antimicrobial activities. The cytotoxicity assessment showed that the P. grandiflora extracts were non-toxic to normal VERO cells. No significant antiviral activity towards Human Herpesvirus type 1 was observed (62 µg/mL). Among the tested varieties, the purple one showed anticancer selectivity towards colon carcinoma cells (RKO).

Betacyanins are a group of water-soluble red-violet compounds containing nitrogen in their structure. These are biosynthesized in red beetroot ( Beta vulgaris L.), a widely consumed vegetable that contains significant amounts of nutritious and bioactive compounds which are also found in dietary supplements. This contribution presents results of betacyanin thermal oxidation (resulting in dehydrogenation) interrelated with decarboxylation in selected acetate/phosphate buffers at pH 3-8 and at 85 °C, which may be of particular significance for formulation and performance of foods. Most of the reaction products were detected at the highest concentrations in the acidic solutions (pH 3-4). The main dehydrogenation reaction pathways were monitored by LC-DAD-MS/MS and were associated with decarboxylation of the principal extract pigments, betanin/isobetanin and neobetanin, at carbon positions C-2 and C-17. Additional reactions are accompanied by the 2,15-decarboxylation processes at different dehydrogenation levels with 15-decarboxy-betanin and 2,15-bidecarboxy-betanin, structurally elucidated by NMR analysis, as the distinct indicators of this route type. For other novel pigments detected, 2,15-bidecarboxy-xanbetanin, 2,15-bidecarboxy-xanneobetanin and 2,15,17-tridecarboxy-neobetanin, additional high resolution mass spectrometric analyses were performed and confirmed their molecular formulas.

Coffee cherry is a rich source of chlorogenic acids (CGAs) and caffeine. In this study we examined the potential antioxidant activity and enzyme inhibitory effects of whole coffee cherries (WCC) and their two extracts on α-amylase, α-glucosidase and acetylcholinesterase (AChE) activities, which are targets for the control of diabetes and Alzheimer's diseases. Whole coffee cherry extract 40% (WCCE1) is rich in chlorogenic acid compounds, consisting of a minimum of 40% major isomers, namely 3-caffeoylquinic acids, 4-caffeoylquinic acids, 5-caffeoylquinic acids, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, 4-feruloylquinc acid, and 5-feruloylquinc acid. Whole coffee cherry extract 70% (WCCE2) is rich in caffeine, with a minimum of 70%. WCCE1 inhibited the activities of digestive enzymes α-amylase and α-glucosidase, and WCCE2 inhibited acetylcholinesterase activities with their IC 50 values of 1.74, 2.42, and 0.09 mg/mL, respectively. Multiple antioxidant assays-including DPPH, ABTS, FRAP, ORAC, HORAC, NORAC, and SORAC-demonstrated that WCCE1 has strong antioxidant activity.

Leafy green vegetables contain essential nutrients and are frequently consumed for their perceived health benefits. The purpose of this study was to profile the nutritional and natural bioactive phytochemical compounds extracted from freeze-dried spinach and kale and compare them with our previously published freeze-dried purslane results. Novel research suggests that these leafy greens contain an abundance of fatty acids, amino acids, organic acids, vitamins, and minerals, which can reduce the risk of many chronic diseases. LC-MS/MS screening identified 69 and 103 compounds in spinach and kale, respectively, including flavonoids, glucosinolates, and phenolic and organic acids. Out of a total of 26 flavonoids identified in the current study, only three were found in spinach. All three leafy greens showed nutritional and health benefits and the unique phytochemical compounds found only in purslane provide a strong basis to incorporate its distinct dietary benefits.

The current work assessed the phytochemical contents of Arabica whole coffee cherry (WCC) and its two commercially available extracts: a minimum 40% chlorogenic acid (CGA; WCCE-1) and 70% caffeine (WCCE-2). Mass spectrometry analyses tentatively identified 219 phytochemicals in the three coffee samples, which is, so far, the largest number of identifications in a single study. A new group of CGA derivative namely caffeoylvaleroylquinic acid (CVQA) was identified in the three samples. Moreover, three 5-hydroxytryptamide derivatives (C 20 -5HT, C 22 -5HT, and C 24 -5HT) were identified in WCC but not in the extracts. Two groups of atractyligenin derivatives (carboxyatractyligenin and noncarboxyatractyligenin) were identified in the three samples. Furthermore, our results show that both extracts retained a large number of phenolic and other potentially bioactive compounds that were naturally present in whole coffee cherry (WCC).

Neutrophils produce hypochlorous acid (HOCl) as well as other reactive oxygen species as part of a natural innate immune response in the human body; however, excessive levels of HOCl can ultimately be detrimental to health. Recent reports suggest that betacyanin plant pigments can act as potent scavengers of inflammatory factors and are notably effective against HOCl. Comparison of the in vitro anti-hypochlorite activities of a novel betalain-rich red beetroot ( Beta vulgaris L.) extract with its pure betalainic pigments revealed that the extract had the highest anti-hypochlorite activity, far exceeding the activity of all of the betalainic derivatives and selected reference antioxidants. This suggests that it may be an important food-based candidate for management of inflammatory conditions induced by excessive HOCl production. Among all pigments studied, betanidin exhibited the highest activity across the pH range.

Purslane (Portulaca oleracea) is a weed naturally found in driveways, lawns, and fields and edible in many regions of Europe, Asia, the Middle East, Africa, and Australia. The purpose of this study was to compare the nutritional and phytochemical components of cultivated and wild purslane. Omega-3 contents of both purslane genotypes were comparable with 189.16 ± 25.52 mg/100 g dry weight and 188.48 ± 6.35 mg/100 g dry weight in cultivated and wild purslane leaves, respectively. Omega-6/omega-3 ratio (1:1-1:3) were low in both genotypes. However, high levels of oxalic acid were observed. Cultivated contained greater amounts of amino acids and vitamins than wild purslane. Of the 184 compounds identified in both genotypes by LC-MS/MS, including phenolic acids, organic acids, flavonoids, alkaloids, and betanin, more than 80 showed greater than two-fold abundance in the wild compared to cultivated purslane. Purslane has the potential to be cultivated as a food ingredient for nutraceutical applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Hypochlorous acid (HOCl) produced by neutrophils is a part of the natural innate immune response system in the human body, but excessive levels of HOCl can ultimately be detrimental to health. Recent reports suggest that betacyanin plant pigments can act as potent scavengers of inflammatory factors and are notably effective against HOCl. In this contribution, chlorination mechanism and position of the electrophilic substitution in betacyanins was studied by high-resolution mass spectrometry and further structural analyses by NMR techniques, which completed the identification of the chlorinated betacyanins. For the study on the influence of the position of decarboxylation on the chlorination mechanism, a comparison of the chlorination position between betanin as well as 17-, and 2,17-decarboxylated betanins was performed. The structural study confirmed that the chlorination position in betanin occurs within the dihydropyridinic moiety at carbon C-18. Therefore, out of the aqueous free chlorine equilibrium species: HOCl, OCl - , Cl 2 , and Cl 2 O, the most potent chlorinating agents are HOCl and Cl 2 O postulated previously and the attack of the Cl ⁺ ion on the carbon C-18 with a cyclic intermediate version is considered., Competing Interests: The authors declare no conflict of interest.

The effects of hot-air drying (AD), freeze drying (FD) and Refractance Window drying (RWD) on the retention of anthocyanins, phenolics, flavonoids, vitamin C and B, and antioxidant capacity (ORAC) in blueberries, tart cherries, strawberries, and cranberries, as well as organic acids and proanthocyanins in cranberries and chlorogenic acid and catchins in blueberries were evaluated. Changes in color, glass transition, specific heat, and surface morphology of the dried fruits were also evaluated. The quality retention in the dried fruits was found to be product and drying method specific. The FD products exhibited a better retention of vitamin C, ORAC, anthocyanins, and phenolics than the AD and RWD fruits. The RWD produced samples with a less effective ORAC, anthocyanins, and phenolics retention, but a higher total vitamin B retention compared to the FD products. The AD-dried fruits displayed significantly lesser quality retention in most of the quality indexes measured in the study., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The determination of residual solvents is critical for quality control in food production and manufacturing processes. A gas chromatographic and mass spectrometric detection (MSD) method was developed and validated to complement the static headspace flame ionization detection (FID) technique for identification, confirmation, and quantitation of solvent residues in botanicals, using Coffeeberry extract and pomegranate powder as test samples. Relative standard deviations (RSDs) of less than 12% were obtained for residual solvents in water and dimethyl sulfoxide. Coffeeberry extract supplemented with10 μg/g of Residual Solvents Class 3 Mix showed MSD and FID recoveries mainly of 91 to 121% and 77 to 110%, respectively, while those samples supplemented with 100 μg/g of Residual Solvents Class 3 Mix displayed MSD and FID recoveries of 105 to 123% and 87 to 112%, respectively. For pomegranate samples supplemented with 10 μg/g of Residual Solvents Class 3 Mix, MSD and FID recoveries of 95 to 124% and 72 to 151% were observed, respectively. Those samples supplemented with 100 μg/g of Residual Solvents Class 3 Mix exhibited MSD and FID recoveries of 109 to 135% and 97 to 127%, respectively. Thus, the developed procedure was demonstrated to be suitable for the analysis of residual solvents in natural food ingredients and is expected to find numerous industrial applications (e.g., process control and impurity detection in final products).

Antioxidants are substances that prevent oxidation of other compounds or neutralize free radicals. Spices and herbs are rich sources of antioxidants. They have been used in food and beverages to enhance flavor, aroma and color. Due to their excellent antioxidant activity, spices and herbs have also been used to treat some diseases. In this review article, the chemical composition and antioxidant activity of spices and culinary herbs are presented. The content of flavonoids and total polyphenols in different spices and herbs are summarized. The applications of spices and their impacts on human health are briefly described. The extraction and analytical methods for determination of antioxidant capacity are concisely reviewed., Competing Interests: The authors declare no conflict of interest.

The paper describes a new, simple, selective and precise high-performance thin-layer chromatographic (HPTLC) method for the simultaneous identification and quantitative determination of boric acid (BA) and calcium fructoborate (CFB) in bulk and tablet/capsule dosage forms of dietary supplements. HPTLC silica gel G 60 F 254 precoated glass plates were used as the stationary phase. The mobile phase consisted of 2-propanol-water 8:2 (v/v). The two boron-based compounds were adequately separated with the R f values of 0.83 ± 0.01 (BA) and 0.59 ± 0.01 (CFB).

A branded calcium fructoborate product, a nature-identical calcium salt of bis (fructose) ester of boric acid found in plants and a natural source of boron in the human diet and sold under the trade name FruiteX-B(®) Brand Calcium Fructoborate ("FrxB"), was evaluated in a 90-day dietary toxicity study and two genotoxicity studies. In the 90-day study, four groups of 10 male and 10 female Crl:SD CD(®) IGS rats were fed diets with FrxB admixtures of 0.56, 1.12, and 1.68% dietary concentration, providing mean overall daily intakes of FrxB in male rats of 385.8, 774.9, and 1161.3 mg/kg bw/day, and 392.1, 784.4, and 1171.1 mg/kg bw/day in female rats. There were no mortalities, no clinical or ophthalmologic signs, body weight, body weight gain, food consumption, food efficiency, Functional Observational Battery (FOB), or Motor Activity (MA) findings associated with the administration of FrxB. There were no adverse changes in hematology, coagulation, clinical chemistry, or urinalysis parameters in male or female rats considered the result of test substance administration. At necropsy, there were no macroscopic, histopathological findings, or organ weight changes deemed related to administration of the test substance. Under the conditions of this study, based on the toxicological endpoints evaluated, the no-observed-adverse-effect level (NOAEL) for FrxB in the diet was 1161.3 and 1171.1 mg/kg bw/day in male and female rats, respectively. Bacterial mutagenicity studies and a micronucleus test using Chinese hamster V79 cells demonstrated no mutagenic or genotoxic potential of the tested brand of calcium fructoborate., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

ORAC and other in vitro methods have to date proved useful in measuring antioxidant potential in foods. In order to better understand the potential relationship between diet and free radical production/mitigation, an in vivo analytic method can provide new insight into directly measuring reactive oxidant species (ROS). Dihydrorhodamine-6G (DHR6G) is indiscriminate to the various free radicals found in humans, and therefore can be useful in quantifying total ROS in vivo. Our aim was to investigate whether the total ROS in human subjects can be quantified using DHR6G after intake of a blend of antioxidants-rich fruit and vegetable-based materials. Twelve participants were given 100 mg of a proprietary blend of fruit, vegetable, and herb powders and concentrates commercially marketed under the trade name "Spectra™". Blood samples were collected at 0, 60, 120 and 180 min and were subsequently tested for ROS in serum using DHR6G as a fluorescent probe. By quantifying this fluorescence, we were able to measure ROS concentrations in human blood. This method is both reliable and efficient for evaluating the efficacy of antioxidants against ROS in vivo. Our data indicate that eleven participants responded to the intake of Spectra™ by significant decreases of ROS concentrations.

This review summarizes published information concerning the determination of antioxidant activity (AA) in coffee samples by various methods (ORAC, FRAP, TRAP, TEAC, etc.) in vitro and limited data of antiradical activity of coffee products in vitro and in vivo. Comparison is carried out of the AA of coffee Arabica and coffee Robusta roasted at different temperatures as well as by different roasting methods (microwave, convection, etc.). Data on the antiradical activity of coffee is provided. The antioxidant activity of coffee, tea, cocoa, and red wine is compared. At the end of this review, the total antioxidant content (TAC) of coffee samples from 21 coffee-producing countries as measured by an amperometric method is provided. The TAC of green and roasted coffee beans is also compared.

The present single-dose study was performed to assess the effect of whole coffee fruit concentrate powder (WCFC), green coffee caffeine powder (N677), grape seed extract powder (N31) and green coffee bean extract powder (N625) on blood levels of brain-derived neurotrophic factor (BDNF). Randomly assorted groups of fasted subjects consumed a single, 100mg dose of each material. Plasma samples were collected at time zero (T0) and at 30 min intervals afterwards, up to 120 min. A total of two control groups were included: subjects treated with silica dioxide (as placebo) or with no treatment. The collected data revealed that treatments with N31 and N677 increased levels of plasma BDNF by about 31% under these experimental conditions, whereas treatment with WCFC increased it by 143% (n 10), compared with baseline. These results indicate that WCFC could be used for modulation of BDNF-dependent health conditions. However, larger clinical studies are needed to support this possibility.

A comprehensive nonenzymatic oxidation mechanism in betanin plant pigment as well as its derivatives, 2-decarboxybetanin, 17-decarboxybetanin, 2,17-bidecarboxybetanin, and neobetanin, in the presence of ABTS cation radicals was investigated by LC-DAD-ESI-MS/MS. The main compounds formed during the first step of betanin and 2-decarboxybetanin oxidation are 2-decarboxy-2,3-dehydrobetanin and 2-decarboxyneobetanin, respectively. In contrast to betanin, the reaction mechanism for 2-decarboxybetanin includes more oxidation pathways. Parallel transformation of 2-decarboxybetanin quinone methide produces neoderivatives according to an alternative reaction that omits the presumably more stabile intermediate 2-decarboxy-2,3-dehydrobetanin. The main oxidation product after the first reaction step for both 17-decarboxybetanin and 2,17-bidecarboxybetanin is 2,17-decarboxy-2,3-dehydrobetanin. This product is formed through irreversible decarboxylation of the 17-decarboxybetanin quinone methide or by oxidation of 2,17-bidecarboxybetanin. Oxidation of neobetanin results primarily in a formation of 2-decarboxy-2,3-dehydroneobetanin by a decarboxylative transformation of the formed neobetanin quinone methide. The elucidated reaction scheme will be useful in interpretation of redox activities of betalains in biological tissues and food preparations.

The antioxidative mechanism of action of betalains is of significant interest because these pigments are recently emerging as highly bio-active natural compounds with potential benefits to human health. Betanidin, the basic betacyanin, comprises the 5,6-dihydroxyl moiety, which results in its high antioxidant activity. Oxidation of betanidin by voltammetric techniques and chro matographic identification of the oxidation products with spectrophotometric and mass spectrometric detection (LC-DAD-MS/MS) were performed. Two main oxidation peaks for betanidin are observable at pH 3-5. These peaks become merged at higher pH, suggesting a different mechanism of oxidation at higher and lower pH values. The low oxidation potential of betanidin confirms its very strong reduction properties. The presence of two prominent oxidized products, 2-decarboxy-2,3-dehydrobetanidin and 2,17-bidecarboxy-2,3-dehydrobetanidin, indicates their generation through two reaction routes with two different quinonoid intermediates: dopachrome derivative and quinone methide. Both lead to the decarboxylative dehydrogenation of betanidin. Subsequent oxidation and rearrangement of the conjugated chromophoric system results in formation of 14,15-dehydrogenated derivatives.

The fruit of the coffee plant, Coffea arabica, has high phenolic antioxidant and phytonutrient content and could be a beneficial food ingredient. However, the fruit has historically been discarded for the favored harvesting of the coffee bean alone. CoffeeBerry products are derived from the whole fruit and include a ground whole powder, a water extract, and a more recently developed water-ethanol extract. The safety of CoffeeBerry products was evaluated in three genotoxicity studies, three short-term oral toxicity studies, and a 90-day dietary toxicity study. Bacterial mutagenicity studies and a micronucleus test using murine peripheral cells demonstrated that none of the three products showed mutagenic or genotoxic potential. In the short-term studies, despite palatability issues, female rats showed a tolerance for whole powder and ethanol extract at doses up to 8800 mg/kg bw/day. Male rats also exhibited palatability issues and tolerated lower doses of approximately 4000 mg/kg bw/day ethanol extract via gavage and approximately 2100 mg/kg bw/day whole powder or water extract in the diet. When fed in the diet to Sprague-Dawley rats for 90 days, ethanol extract showed no adverse effects at dietary concentrations of up to 5% (approximately 3446 and 4087 mg/kg bw/day for male and female rats, respectively)., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)