Determination of the relative contribution of quercetin and its glucosides to the antioxidant capacity of onion by cyclic voltammetry and spectrophotometric methods

This paper describes the use of cyclic voltammetry (CV), spectrophotometric methods [Trolox equivalent antioxidant capacity (TEAC), peroxyl radical trapping capacity (PRTC), DPPH radical scavenging activity (RSA), and Folin-Ciocalteu reagent (FCR) reducing capacity], and photochemiluminescence (PCL) for the measurement of the antioxidant capacity of onion var. Sochaczewska and var. Szalotka. The antioxidant and reducing activity of the dominant onion flavonoids quercetin (Q), quercetin-3- O-beta-glucoside (Q3G), quercetin-4'- O-beta-glucoside (Q4'G), and quercetin-3,4'-di- O-beta-glucoside (Q3,4'G) were determined by spectrophotometric (TEAC and PRTC) and CV methods, respectively. The contribution of quercetin and its glucosides to the antioxidant capacity of onion was calculated in consequence of the qualitative and quantitative analysis of onion flavonoids by high-performance liquid chromatography-ultraviolet-mass spectrometry. The dominant forms of quercetin in the onion var. Sochaczewska and Szalotka included Q4'G (61 and 54%), Q3,4'G (37 and 44%), Q3G (1.4 and 1.1%), and free quercetin (1.1 and 0.7%), respectively. The CV experiment showed the highest reducing activity of Q while Q3G, Q4'G, and Q3,4'G exhibited about 68, 51, and 30% of the reducing power noted for Q. The order of the reducing activity of onion flavonoids was confirmed by their free radical scavenging activity and evaluated by TEAC and PRTC assays as follows: Q > Q3G > Q4'G > Q3,4'G. The Q4'G and Q3,4'G showed poor antioxidant activity under both applied spectrophotometric assays but still exhibited reducing activity based on CV experiments. The reducing capacity of onions determined by CV method was twice higher than the antioxidant capacity formed by water-soluble compounds (ACW) evaluated by PCL, and it was about 50% higher than PRTC and DPPH RSA results and the converted FCR reducing capacity. In contrast, the reducing capacity of onions determined by the CV method was 3-fold and about four times lower when compared to the antioxidant capacity evaluated by the TEAC method and that formed by lipid-soluble compounds (ACL) provided by PCL, respectively. The highest antioxidant capacity of onion was found under cumulative consideration of PCL (ACW + ACL) and TEAC assays. The relative contribution of Q and its glucosides to the antioxidant capacity of onions showed a low contribution of Q, Q3G, and Q3,4'G derived from CV, TEAC, and PRTC assays while the highest contribution to the antioxidant capacity of onions was provided by Q4'G.     

Uptake of quercetin and quercetin 3-glucoside from whole onion and apple peel extracts by Caco-2 cell monolayers

Evidence suggests that regular consumption of fruits and vegetables may reduce the risk of chronic diseases, and phytochemicals from fruits and vegetables may be responsible for this health benefit. However, there is limited knowledge on the bioavailability of specific phytochemicals from whole fruits and vegetables. This study used Caco-2 cells to examine uptake of quercetin aglycon and quercetin 3-glucoside as purified compounds and from whole onion and apple peel extracts. Pure quercetin aglycon was absorbed by the Caco-2 cells in higher concentrations than quercetin 3-glucoside (p < 0.05). Caco-2 cells treated with quercetin 3-glucoside accumulated both quercetin 3-glucoside and quercetin. Caco-2 cells absorbed more onion quercetin aglycon than onion quercetin 3-glucoside (p < 0.05), and the percentage of onion quercetin absorbed was greater than that of pure quercetin, most likely due to enzymatic hydrolysis of quercetin 3-glucoside and other quercetin glucosides found in the onion by the Caco-2 cells. Caco-2 cells absorbed low levels of quercetin 3-glucoside from apple peel extracts, but quercetin aglycon absorption was not detected. Caco-2 cell homogenates demonstrated both lactase and glucosidase activities when incubated with lactose and quercetin 3-glucoside, respectively. This use of the Caco2 cell model appears to be a simple and useful system for studying bioavailability of whole food phytochemicals and may be used to assess differences in bioavailability between foods.     

Analysis of the oxidative degradation of proanthocyanidins under basic conditions

Proanthocyanidin isolates from grape (Vitis vinifera L. cv. Pinot noir) skin and seed underwent oxidative degradation in solution (10 g/L) under basic conditions while exposed to atmospheric oxygen. Degradation was monitored by reversed-phase HPLC following acid-catalyzed cleavage in the presence of excess phloroglucinol (phloroglucinolysis) and by high-performance gel permeation chromatography. All isolates degraded under these conditions and followed second-order kinetics for over 1 half-life, consistent with an oxidation reaction. The conversion of proanthocyanidins to known subunits (conversion yield) when measured by phloroglucinolysis showed a dramatic decline over the course of the reaction. With the exception of (+)-catechin extension subunits, all individual subunits decreased in concentration during the oxidation process, also following second-order kinetics for over 1 half-life. Skin proanthocyanidins degraded the fastest due to the presence of (-)-epigallocatechin extension subunits. Seed procyanidins were degraded with and without flavan-3-ol monomers. Flavan-3-ol monomers slowed the rate of seed procyanidin degradation. The mean degree of polymerization (mDP) determined by phloroglucinolysis indicated a large decrease in mDP as the reaction progressed; yet, by GPC, the size distribution of all proanthocyanidins changed little in comparison. The conversion yield could be an important parameter to follow when using phloroglucinolysis as a means for determining proanthocyanidin mDP, and when monitoring the oxidative degradation of proanthocyanidins.     

FTIR-ATR analysis of brewed coffee: effect of roasting conditions

FTIR-ATR was used to study the effect of roasting conditions on the flavor of brewed coffee using Guatemala Antigua coffee beans. The 1800-1680 cm(-1) carbonyl region for vinyl esters/lactones, esters, aldehydes, ketones, and acids was found to provide a flavor-print of the brewed coffee. A study of light, medium, and dark roasts indicated that when the rate of heating to the onset of the first and second cracks was kept constant, the types of carbonyl compounds formed were similar, varying only in their concentration. This difference in concentration is apparently due to the additional heating of the coffee bean beyond the second crack. When the heating rate to the onset of the first and second crack was varied, both the types and concentration of the carbonyl compounds formed during roasting were affected. Thus, heating rates of green coffee beans to the onset of the first and second cracks are important determinants of the basic taste and aroma of brewed coffee.     

Detailed examination of the degradation of phenol derivatives under oxygen delignification conditions

Several phenolic compounds were subjected to oxygen-alkali oxidations under oxygen delignification conditions, and their degradations were examined in detail by applying a novel formula. The formula was established on the basis of the two following considerations. The degradation of the phenolic compounds should be expressed by the sum of two types, each of which is caused by molecular oxygen and by highly reactive active oxygen species (HAOS). The degradation should be described by a mathematical equation to which a rate function, k(t), dependent on reaction time t, is applied instead of a rate constant. By rearrangements, the following formula was obtained: k(t) = A/(t + B) (A, B are constants). This is hyperbola, and the illustration of k(t) visualized the contribution of HAOS to the degradation of the phenolic compounds. HAOS did not contribute so much to the degradation, especially at 70 degrees C, which suggests the low energy supply for the HAOS generation at 70 degrees C. The extrapolation of k(t) to the beginning of the reaction gives its initial value, k(initial), which is the rate constant of the reaction between the phenolic compounds and molecular oxygen. As expected, k(initial) was dependent on the electronic property of their substituents. Quantification of the phenolic compounds degraded by HAOS showed that the contribution of HAOS to the degradation is not great. The maximum contribution was observed in the oxidation of 2,6-dimethylphenol at 85 degrees C. In this case, 5 and 95% of the compound were degraded by HAOS and molecular oxygen, respectively.     

不同贮藏条件下荷花粉脂质氧化与色泽降解动力学模型

为了减少干燥后荷花粉贮藏过程中品质的劣变,该研究选用可真空和避光的包装材料,分别研究了30 d贮藏过程中温度(4、20、30℃)、光照(避光、非避光)和氧气(真空、常压)对荷花粉黄色度值(b*值)、过氧化氢值(peroxide value,POV)和硫代苯巴比妥酸反应底物值(thiobarbituric acid reactive substances,TBArs)变化动力学的影响;同时,采用零阶和一阶数学模型对试验数据进行模拟。结果表明:贮藏温度对荷花粉贮藏过程中品质劣变具有显著影响。在避光真空包装条件下,温度为20和30℃贮藏30 d,与4℃贮藏条件相比,荷花粉黄色度值显著降低(P0.05),POV值和TBArs值显著上升(P0.05);贮藏温度为4℃,真空包装贮藏发现,光照对荷花粉POV值的影响不显著(P0.05),但明显加快了脂质氧化的传播速度,非避光条件下贮藏30 d,TBArs值上升了7.42 mg/kg;零阶模型可较好的预测荷花粉贮藏过程中脂质TBArs值的变化,一阶模型可较好的预测荷花粉贮藏过程中黄色度和脂质POV值的变化规律。研究结果为脱水后荷花粉贮藏过程中品质劣变规律的探索及贮藏条件的选择提供了一定的基础。     

Mobility and degradation of the herbicide imazosulfuron in lysimeters under field conditions

The mobility and degradation of imazosulfuron, labeled with carbon-14 at the imidazole ([imi-(14)C]imazo) or pyrimidine ring ([pyr-(14)C]imazo), in lysimeters with 1 m(2) surface and 110 cm depth were investigated for three years. One lysimeter was treated with [imi-(14)C]imazo in two successive years at the rate of 50 g of active ingredient (ai)/ha each. The other two lysimeters were treated once with [pyr-(14)C]imazo and a mixture (1:1, w/w) of the two labeled imazosulfurons, respectively (50 g of ai/ha). In the first and second years of monitoring, the yearly mean concentration of (14)C in the leachate water was <0.10 microg/L in each lysimeter. Although in the third year the concentration of (14)C in the leachate water was 0.17 microg/L for the lysimeter treated twice with [imi-(14)C]imazo, the concentration of imazosulfuron and its degradation products in the leachate water ranged from 0.01 to 0.06 microg/L. At the study termination, the main portion of (14)C recovered was found in the upper 30 cm soil layer in each lysimeter, and no (14)C was detected below a depth of 50 cm. These findings indicated that imazosulfuron and its degradation products in soils translocated into groundwater only slightly.     

Coffee roasting and aroma formation: application of different time-temperature conditions

The impact of time-temperature combinations of roasting processes on the kinetics of aroma formation in coffee was investigated. The development of 16 aroma compounds and the physical properties of coffee beans was followed in a commercial horizontal drum roasting process and in laboratory scale fluidizing-bed roasting processes at high temperature-short time and low temperature-long time conditions. All trials were run to an equal roast end point as defined by the lightness of coffee beans. In addition, the effect of excessive roasting on aroma composition was studied. Compared to low temperature-long time roasting, high temperature-short time roasting resulted in considerable differences in the physical properties and kinetics of aroma formation. Excessive roasting generally led to decreasing or stable amounts of volatile substances, except for hexanal, pyridine, and dimethyl trisulfide, whose concentrations continued to increase during over-roasting. When the drum roaster and the fluidizing bed roaster were operated in the so-called temperature profile mode, that is, along the identical development of coffee bean temperature over roasting time, the kinetics of aroma generation were similar in both processes.     

Effect of roasting conditions on reduction of ochratoxin a in coffee

A commercial lot of green coffee, naturally contaminated with ochratoxin A (OTA), was roasted under various conditions, and the effects on its final OTA content were determined. Precautions were taken in sampling the coffee to cope with OTA inhomogeneity. The roasting conditions were kept within the range of commercial practice. Roasting time was varied from 2.5 to 10 min, and the roast color varied from light medium to dark. The differences in OTA reduction between the different levels of roasting times and colors did not reach statistical significance. However, for all roasting conditions, the reduction was highly significant, 69% reduction over the combined results. In total, nine studies by various authors about OTA reduction during coffee roasting are now available. Seven out of these nine reported that the relevant range of OTA reductions was between 69 and 96%. Among these seven,are all four studies that reported using naturally contaminated beans, a sampling procedure adapted to mycotoxin inhomogeneity, and roasting conditions within the range of actual practice. Three different explanations are available for this reduction: physical removal of OTA with chaff, isomerization at the C-3 position into another diastereomer, and thermal degradation with possible involvement of moisture. All three explanations may play a partial role in the OTA reduction during coffee roasting.     

Oxidation of quercetin by salivary components. Quercetin-dependent reduction of salivary nitrite under acidic conditions producing nitric oxide

Under acidic conditions, nitrite is protonated to nitrous acid (pK(a) = 3.2-3.4) that can be transformed into nitric oxide by self-decomposition and reduction. When sodium nitrite was mixed with quercetin at pH 1-2, quercetin was oxidized producing nitric oxide. In addition to quercetin, kaempferol and quercetin 4'-glucoside were also oxidized by nitrous acid, but oxidation of apigenin, luteolin, and rutin was slow compared to oxidation of the above flavonols. These results suggested that flavonols, which have a free hydroxyl group at carbon position 3, can readily reduce nitrous acid to nitric oxide. When the pH of saliva was decreased to 1-2, formation of nitric oxide was observed. The nitric oxide formation was enhanced by quercetin, and during this process quercetin was oxidized. These results indicate that there is a possibility of reactions between phenolics and nitrous acid derived from salivary nitrite in the stomach.     

The effect of zeolite,selenium and silicon on qualitative and quantitative traits of onion grown under salinity conditions

Considering the importance of onion and its cultivation in saline soils, a two-year experiment was carried out to investigate the effects of soil applied zeolite and foliar application of selenium (Se) and silicon (Si) on onion yield, qualitative traits and physiological attributes, under salinity stress. An experiment was performed at three-way factorial design with three replications as follows: zeolite at three levels (0, 4 and 8 ton ha-1), Se at three levels (0, 0.5 and 1 kg ha-1) and Si at three levels (0, 200 and 400 kg ha-1). The results indicated that the effect of year was significant on all studied traits except for soluble solids, bulb nitrogen, leaf sodium and potassium, chlorophyll content and peroxidase activity. Number of small-sized bulbs decreased with increasing zeolite, Se and Si application. Dry matter, soluble solids, nitrogen content, nitrate concentration as well as protein content significantly increased due to zeolite, Se and Si application. Leaf sodium content and enzyme activity decreased due to zeolite, Se and Si application. Overall, 8 ton ha-1 zeolite along with 1 kg ha-1 Se and 400 kg ha-1 Si caused the maximum onion yield and qualitative and physiologic traits including soluble solids, potassium, protein, chlorophyll and photosynthesis .     

Effect of culture acclimation on the kinetics of aldicarb insecticide degradation under methanogenic conditions

This study reports on the kinetics of aldicarb transformation under methanogenic conditions using batch reactors containing acclimated and unacclimated cultures under controlled conditions. Culture acclimation was accomplished by exposing anaerobic microorganisms maintained in a semi-batch reactor to low concentrations of aldicarb. Results of the kinetic studies showed that in an anaerobic system aldicarb is converted to aldicarb nitrile by the hydrolytic pathway. Analysis of the hydrolysis/dehydration rate constants showed that anaerobic cultures enhanced the rate of conversion of aldicarb by 4-fold for acclimated cultures and by 2-fold for unacclimated cultures compared to the rate of abiotic hydrolysis (p < 0.05). Only the acclimated cultures were able to further mineralize the reaction intermediate aldicarb nitrile. Michaelis-Menten and Monod kinetics adequately defined the aldicarb nitrile degradation.     

Thermal degradation studies of food melanoidins

Food melanoidins were isolated from bread crust, coffee, and tomato sauce and their composition was investigated by thermal degradation. Among the generated volatiles, important food flavor compounds were detected: in particular furans, carbonyl compounds, 1,3-dioxolanes, pyrroles, pyrazines, pyridines, thiophenes, and phenols. The results indicated that the isolated melanoidin fractions mainly consisted of compounds formed from carbohydrates and their degradation products. Besides proteins, other food constituents were incorporated in the melanoidin structure as well, such as lipid oxidation products in tomato melanoidins and phenolic compounds in coffee melanoidins. A comparison of the thermal generation of volatiles between these food-derived melanoidins and model melanoidins prepared from a single carbonyl compound and an amino acid showed that the degradation pattern of food melanoidins is quite different from that obtained from a glucose-glycine model system.     

Identification and in vitro cytotoxicity of ochratoxin A degradation products formed during coffee roasting

The mycotoxin ochratoxin A is degraded by up to 90% during coffee roasting. In order to investigate this degradation, model heating experiments with ochratoxin A were carried out, and the reaction products were analyzed by HPLC-DAD and HPLC-MS/MS. Two ochratoxin A degradation products were identified, and their structure and absolute configuration were determined. As degradation reactions, the isomerization to 14-(R)-ochratoxin A and the decarboxylation to 14-decarboxy-ochratoxin A were identified. Subsequently, an analytical method for the determination of these compounds in roasted coffee was developed. Quantification was carried out by HPLC-MS/MS and the use of stable isotope dilution analysis. By using this method for the analysis of 15 coffee samples from the German market, it could be shown that, during coffee roasting, the ochratoxin A diastereomer 14-(R)-ochratoxin A was formed in amounts of up to 25.6% relative to ochratoxin A. The decarboxylation product was formed only in traces. For toxicity evaluations, first preliminary cell culture assays were performed with the two new substances. Both degradation products exhibited higher IC50 values and caused apoptotic effects with higher concentrations than ochratoxin A in cultured human kidney epithelial cells. Thus, these cell culture data suggest that the degradation products are less cytotoxic than ochratoxin A.     

Formation of tetracycline degradation products in chicken and pig meat under different thermal processing conditions

Tetracycline (TC) and 4-epitetracycline (4eTC) degradation, as well as anhydrotetracycline (ATC) and 4-epianhydrotetracycline (4eATC) formation, has been evaluated in thermally treated chicken breast, pig loin, and pig loin with added back-fat. Samples containing TC and 4eTC residues were submitted to microwave or boiling heating, extracted with a mixture of McIlvaine buffer/methanol (75:25), and analyzed by high-performance liquid chromatography-diode array detection on a phenyl-hexyl reverse phase chromatographic column. The formation of ATC and 4eATC, as well as of two unidentified compounds, was described for the first time in edible meat samples submitted to mild thermal treatments, similar to those applied at home to cook foods. Degradation of TC and 4eTC and formation of ATC and 4eATC versus time of treatment fitted satisfactorily a first-order kinetic. Even if the potential toxic effects of these breakdown compounds should be further investigated, their formation in cooked meat should be taken into account when maximum residue limits are established.     

Thermal degradation of the Fusarium mycotoxin deoxynivalenol

Deoxynivalenol (DON) is a toxic secondary metabolite produced by molds of the Fusarium genus, which are able to infect cereal crops in the field. Concerning its rate of occurrence and mean concentration, DON is one of the most important mycotoxins in cereal commodities. Its toxic effects range from causing diarrhea, vomiting, and gastro-intestinal inflammation to noncompetitive inhibition of the biosynthesis of proteins in eukaryotic cells. To study the stability of DON under food-processing conditions such as cooking or baking, we performed model heating experiments and screened the residue for degradation products. Heating of DON and 3-acetyldeoxynivalenol (3-AcDON), especially under alkaline conditions, gave a mixture of compounds, which were isolated and structurally elucidated by NMR and MS experiments. Three of these compounds were already known (norDON A, norDON B, and norDON C), while four were new and named 9-hydroxymethyl DON lactone, norDON D, norDON E, and norDON F. The significance of the DON degradation products was checked by analyzing commercially available food samples. norDON A, B, and C were detected in 29-66% of the samples in mean concentrations ranging from 3 to 15 microg/kg. Furthermore, cell culture experiments using IHKE cells showed that the compounds that were detected in food samples are less cytotoxic in the formazan dye cytotoxicity assay compared to DON. Whereas DON revealed a median effective concentration (EC50) at 1.1 micromol/L, all other compounds did not show any significant effect up to 100 micromol/L. These findings indicate that the degradation of DON under thermal treatment might reduce the toxicity of DON contaminated food.     

Quercetin-dependent reduction of salivary nitrite to nitric oxide under acidic conditions and interaction between quercetin and ascorbic acid during the reduction

A salivary component, nitrate, is reduced to nitrite in the oral cavity. Polyphenols in foods are mixed with nitrite in the saliva to be swallowed into the stomach. An objective of the present study is to elucidate reactions between a polyphenol quercetin and a nitrite under acidic conditions. Nitric oxide, which is formed by the reactions between nitrous acid and quercetin or ascorbic acid (AA), can be measured using an oxygen electrode in the saliva as well as a buffer solution. The initial oxidation of quercetin was inhibited by AA, and quercetin enhanced the oxidation of AA, suggesting AA-dependent reduction of quercetin radicals, which might be formed during the oxidation of quercetin by nitrous acid. On the basis of the above results, the usefulness of an oxygen electrode for the measurement of nitrite-dependent nitric oxide formation under acidic conditions is proposed and the possible mechanism of reduction of nitrous acid by quercetin and the interaction between quercetin and AA, which is a normal component in the gastric juice, for the reduction of nitrous acid is discussed.     

Thermal degradation studies of glucose/glycine melanoidins

Nondialyzable and water-insoluble melanoidins, isolated from a glucose/glycine model reaction mixture, which was prepared in a standardized way according to the guidelines of the COST Action 919, were heated at different temperatures ranging from 100 to 300 degrees C. Among the volatile compounds, which were analyzed by SPME and GC-MS, pyrazines, pyridines, pyrroles, and furans were detected. In general, total amounts of volatile compounds increased with the temperature. When water-insoluble melanoidins were heated, especially at higher temperatures, this resulted in a higher diversity of isolated compounds. For furans, pyrroles, pyrazines, and carbonyl compounds a maximum was observed in the case of high molecular weight melanoidins around 200-220 degrees C. Pyridines and total oxazoles, however, were generated in higher yields with increasing temperatures. These results demonstrate the possibility of producing some flavor-significant volatiles from heated standard melanoidins at temperatures relevant to food preparation and contribute to the flavor aspects originating from melanoidins.     

Thermal degradation of commercial grade sodium copper chlorophyllin

Sodium copper chlorophyllin (SCC), a water-soluble commercial derivative of chlorophyll, has gained importance as a food colorant and dietary supplement with apparent chemopreventive activities. The thermal stability of SCC was studied to assess the potential application of this chlorophyll derivative for use in thermally processed foods and supplements. Thermal degradation of an aqueous 500 ppm SCC solution was monitored between 25 and 100 degrees C by a loss of absorbance at 627 nm. Decomposition was also followed by reversed phase C18 HPLC with photodiode array detection to monitor the loss of Cu(II)Chlorin e4, the major component of commercial grade SCC. The rate of thermal degradation of SCC was found to follow first-order reaction kinetics. HPLC analysis confirmed the ultraviolet and visible absorbance data and also demonstrated loss of the major SCC component, Cu(II)Chlorin e4, at a rate faster than that of overall SCC. The activation energy was estimated using the Arrhenius equation and found to be 13.3 +/- 0.8 and 16.0 +/- 2.1 kcal/mol for the thermal degradation of SCC and Cu(II)Chlorin e4, respectively. The observed temperature sensitivity of SCC was determined to be similar to that of natural chlorophyll and raises the possibility of color deterioration when used in food products where mild to severe thermal treatment is applied. Furthermore, the implication of rapid loss of Cu(II)Chlorin e4, a reported bioactive component of SCC, upon heating may result in alteration of potential dietary benefits such as antimutagenic and antioxidant activity.     

Thermal degradation of sulforaphane in aqueous solution.

Sulforaphane, a cancer chemopreventive agent identified from broccoli, was degraded in an aqueous solution at 50 and 100 degrees C. The reaction mixtures were extracted with methylene chloride and analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). Dimethyl disulfide, S-methyl methylthiosulfinate, S-methyl methylthiosulfonate, methyl (methylthio)methyl disulfide, 1,2,4-trithiolane, 4-isothiocyanato-1-(methylthio)-1-butene, and 3-butenyl isothiocyanate were identified as volatile decomposition products. After methylene chloride extraction, the aqueous layer was dried and silica gel column chromatography was used to separate and purify the nonvolatile decomposition products. The major thermal degradation compound was determined by (1)H NMR, (13)C NMR, and FAB-MS as N, N'-di(4-methylsulfinyl)butyl thiourea. A possible mechanism for the formation of these products is proposed.