Total Phenolic and Phytosterol Compounds and the Radical Scavenging Activity of Germinated Australian Sweet Lupin Flour

In addition to their favourable nutritional profile, legumes also contain a range of bioactive compounds such as phenolic compounds and phytosterols which may protect against chronic diseases including cancer and cardiovascular disease. Germination of some legume seeds has been previously reported to increase the concentration of the bioactive compounds. In this study, the effect of germination of Australian Sweet Lupin (ASL) seeds for 9 days on the concentration of some bioactive compounds and the radical scavenging activity in the resulting flour was determined. The concentration of total phenolic compounds in methanolic extracts of germinated ASL flour was determined using Folin Ciocalteu reagent and phytosterols in oil extracts were analyzed by gas–liquid chromatography. The methanolic and oil extracts were also used to determine radical scavenging activity toward 2,2-diphenyl-1-picrylhydrazyl. In the methanolic extracts of germinated ASL flour, phenolic contents and the antioxidant activity were significantly increased following germination (700 and 1400 %, respectively). Analysis of the oil extracts of germinated ASL flour revealed that the concentration of phytosterols and the antioxidant activity were also increased significantly compared to ungerminated ASL flour (300 and 800 %, respectively). The relative proportion of phytosterols in germinated ASL flour was: β-sitosterol (60 %), stigmasterol (30 %) and campesterol (10 %). Germination increases the concentration of bioactive compounds and the radical scavenging activity in the germinated ASL flour.     

超声波和微波对柑橘杂交种果皮中酚类物质浸提及其抗氧化能力的影响

以柑橘杂交品种葡萄柚、茂谷橘橙果皮为材料,研究超声波和微波对柑橘果皮提取物中酚类物质及其抗氧化活性的影响。结果表明:葡萄柚果皮中含有丰富的柚皮苷、芦丁、橙皮素等酚类物质,茂谷橘橙果皮中含有丰富的槲皮素、川陈皮素、橙皮素等酚类物质。葡萄柚果皮提取液的抗氧化活性高于茂谷橘橙。葡萄柚果皮超声波提取液中酚类物质的含量和抗氧化活性高于微波提取液。柑橘果皮提取物中酚类物质的含量与其DPPH自由基和羟自由基清除能力呈正相关。此结果说明超声波提取法可提高葡萄柚果皮中酚类物质的提取率和提取液抗氧化能力。     

Black currant polyphenols: Their storage stability and microencapsulation

Interest in the consumption of fresh fruits is, to a large extent, due to its content of bioactive nutrients and their importance as dietary antioxidants. There is a growing demand for delivery of antioxidants through functional foods with the related challenge of protecting their bioactivity during food processing and subsequent passage through the gastrointestinal tract. This study focuses on the evaluation of concentration of bioactive compounds in black currant berries (Ribes nigrum L.) and retention of black currant polyphenol compounds and their antioxidant activity after microencapsulation by spray-drying. The polyphenol contents and antioxidant activities were assessed for 5 black currant cultivars grown in Canada in fresh and stored fruits at −20 °C for 9 months. The Ben Alder cultivar was the richest in anthocyanins and flavonols content with high antioxidant potential measured with ABTS and DPPH radicals (4.5 and 2.4 mM/100 g). Black currant polyphenols were stable during 9 month of storage at −20 °C. The results of this study indicated that in black currants storage did not affect negatively the antioxidant capacity. The good stability of bioactive compounds of black currants during freezer storage allows prolonging the storage time of these berries. Maltodextrins with different DE (dextrose equivalent): DE11, DE18, DE21 and inulin were selected as wall material. The stability of encapsulated polyphenols during 12 months storage at 8 °C and 25 °C was evaluated. Compared to maltodextrins DE18 and DE21, maltodextrin DE11 had not only higher drying yield but also offered better protection for phenolics during storage. Radical scavenging activity studies demonstrated significant antioxidant activity of microencapsulated powders before and after storage. The black currant polyphenol microcapsules described in this study represent a promising food additive for incorporation into functional foods due to antioxidant content.     

The Content of Phenolic Compounds and Radical Scavenging Activity Varies with Carrot Origin and Root Color

The contents of phenolic compounds and radical scavenging activities were assessed in a carrot collection comprising 35 cultivars, landraces and breeding populations. The accessions originated from various world regions and they represented Eastern and Western carrot gene pools. In two-year field trial carrot roots of orange, red, yellow, white and purple color were cultivated, freeze-dried and analyzed for phenolic content by Folin-Ciocalteu assay and UV/Vis assay. Radical scavenging activity in the extracts was determined with a stable DPPH radical. Carrots developing purple roots possessed on average 9 times more phenolics than roots of other colors. Furthermore, they were rich in anthocyanins that caused very high antiradical activity. Red carrots showed higher antioxidant activity than orange, yellow and white carrots and in the season of lower rainfall they accumulated higher amounts of phenolic compounds. Carrots of Asian origin belonging to Eastern gene pool were more often purple or red and richer in phenolics and had higher antiradical activity than those from the Western gene pool with mainly orange roots.     

Hydrothermal and biotechnological treatments on nutraceutical content and antioxidant activity of rice bran

The effect of processing such as steaming, germination and parboiling on nutraceuticals and in vitro bioactive properties of rice bran from three different rice varieties namely Jyothi (pigmented), IR64 and Sona masuri (non-pigmented) were investigated. Within the varieties envisaged, pigmented Jyothi variety contained higher levels of vitamin E, soluble, bound and total polyphenol, flavanoids, free radical scavenging activity and total antioxidant activity. Direct steam exposure of bran resulted in an increase in, ether extractives and oryzanol, as well as retention of all the vitamin E components, bound polyphenols, flavonoids and decrease in soluble and total polyphenol content, free radical scavenging activity and total antioxidant activity compared to native. Parboiling as well as germination of paddy resulted in an increase, in the content of ether extractives and oryzanol, whereas other bioactive properties decreased compared to native. Hence it may be concluded that bioactive components and antioxidant properties were significantly higher in Jyothi bran compared to the other two paddy brans, and processing leads to changes in bioactive properties with maximum retention of bioactive components in the steamed bran.     

Carotenoids,Phenolic Compounds and Tocopherols Contribute to the Antioxidative Properties of Some Microalgae Species Grown on Industrial Wastewater

This study aimed at investigating the potential of microalgae species grown on industrial waste water as a new source of natural antioxidants. Six microalgae from different classes, including Phaeodactylum sp. (Bacillariophyceae), Nannochloropsis sp. (Eustigmatophyceae), Chlorella sp., Dunaniella sp., and Desmodesmus sp. (Chlorophyta), were screened for their antioxidant properties using different in vitro assays. Natural antioxidants, including pigments, phenolics, and tocopherols, were measured in methanolic extracts of microalgae biomass. Highest and lowest concentrations of pigments, phenolic compounds, and tocopherols were found in Desmodesmus sp. and Phaeodactylum tricornuotom microalgae species, respectively. The results of each assay were correlated to the content of natural antioxidants in microalgae biomass. Phenolic compounds were found as major contributors to the antioxidant activity in all antioxidant tests while carotenoids were found to contribute to the 1,1-diphenyl-2-picryl-hydrazil (DPPH) radical scavenging activity, ferrous reduction power (FRAP), and ABTS-radical scavenging capacity activity. Desmodesmus sp. biomass represented a potentially rich source of natural antioxidants, such as carotenoids (lutein), tocopherols, and phenolic compounds when cultivated on industrial waste water as the main nutrient source.     

Effect of Genotype on the Sprouting of Pomegranate (<Emphasis Type="Italic">Punica granatum</Emphasis> L.) Seeds as a Source of Phenolic Compounds from Juice Industry by-Products

Pomegranate (Punica granatum L.) fruits are used mainly by the juice industry, for which seeds are a by-product to be disposed of, though they could potentially be a source of bioactive compounds. In this work, germination (total germination percentage, G; mean germination time, MGT; time to reach 80% of germination, TG80; seedling shoot length, fresh weight and dry matter), and nutritional value (total phenolics, TP; total flavonoids, TF; total non-tannins, TNT; antioxidant activities) of pomegranate seeds and sprouts were determined on four commercial pomegranate cultivars (Akko, Dente di Cavallo, Mollar de Elche and Wonderful). Seeds were removed from ripe fruits and incubated in plastic trays containing sterile cotton wetted with distilled water. Sprout shoots were harvested when they reached the complete cotyledon expansion, i.e., the ready-to-eat stage. Akko showed the best germination performance (G = 98%; MGT = 14 days after sowing, DAS; TG80 = 16 DAS), followed by Mollar de Elche. Sprouting dramatically increased TP, TF, TNT and antioxidant activity in all genotypes, with the highest values recorded in Mollar de Elche and Dente di Cavallo. Overall, based on germination performance, Akko and Mollar de Elche would be the best cultivars for sprouting. Sprouting pomegranate seeds appears to be a suitable way of utilizing by-products of the juice industry to obtain bioactive compounds.     

Antioxidant activity of small grain cereals caused by phenolics and lipid soluble antioxidants

In this study, the content of soluble, free forms of phenolic compounds (total phenolics, flavonoids, PVPP (polyvinylpolypyrrolidone) bound phenolics, proanthocyanidins and phenolic acids), as well as the content of carotenoids and tocopherols, were determined in whole grains of bread and durum wheat, rye, hull-less barley and hull-less oat, each represented with four genotypes. Antioxidant activity was evaluated as radical scavenging activity with DPPH (2,2-diphenyl-1-picrylhydrazyl) reagent, as well as by hydrogen transfer reaction (reduction power) based on the reduction of Fe³⁺. Generally, a considerable variation in antioxidant activities and phytochemical contents was observed between the cereals. Remarkably higher DPPH radical scavenging ability and reducing power were detected in hull-less barley, followed by rye and hull-less oat and durum and bread wheat, indicating that small grain species have different major antioxidants with different properties. Hull-less barley had the highest content of total free phenols, flavonoids, PVPP bound phenolics and contained flavan-3-ols, not found in other species. Hull-less oat had the highest content of tocopherols, very high content of yellow pigments and PVPP bound phenolics. Ferulic acid was the major free phenolic acid in small grain cereals tested. The relationship between the content of soluble phenols, as well as reducing power and DPPH^• scavenging activity are also considered.     

Nutritional and Pharmaceutical Properties of Bioactive Compounds in Organic and Conventional Growing Kiwifruit

The bioactivity of two kiwifruit’s cultivars growing under organic and conventional conditions were studied and compared. The bioactive compounds were extracted with water and ethanol using similar conditions which are applied in pharmaceutical applications and for daily fruit consumption such as tea drink. Antioxidant radical scavenging assays [ferric-reducing/antioxidant power (FRAP); cupric reducing antioxidant capacity (CUPRAC); 2, 2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)], fourier transform infrared (FT-IR) and ultraviolet spectroscopy, two (2D-FL) and three-dimensional (3D-FL) fluorometry were used for the detection of biologically active metabolites derived from kiwifruits (total phenols, flavonoids, chlorophylls, carotenoids and ascorbic acid). The correlation between the total phenol content (TPC) and other bioactive compounds, and their total antioxidant capacities (TAC) was calculated for studied kiwifruit’s extracts. The interaction between drugs and human serum albumin (HSA) plays an important role in the distribution and metabolism of drugs. The properties of kiwifruit’s phenol extracts showed their ability to quench HSA, forming the complexes similar to the ones between the proteins and pure flavonoids such as quercetin. The cultivar ‘Bidan’ exhibited significantly higher TAC than the classic ‘Hayward’. In conclusion, for the first time ‘Bidan’ organic kiwifruit was analyzed and compared with widely consumed ‘Hayward’, using its bioactive and fluorescence properties. The influence of physiologically active kiwifruit’s compounds on human health, through our investigations in vitro and scientifically proven information, was explained. Relatively high content of bioactive compounds, high antioxidant and fluorescence properties of kiwifruit justify its use as a source of valuable antioxidants.     

Evaluation of the Total Antioxidant Capacity,Polyphenol Contents and Starch Hydrolase Inhibitory Activity of Ten Edible Plants in an In vitro Model of Digestion

The total phenolics contents, total antioxidant capacity (TAC) and starch hydrolase inhibitory activity of the aqueous extracts of 10 edible plants and the stability of these parameters after the gastric and duodenal digestion in an in vitro model was investigated. The TAC was evaluated using the oxygen radical absorbance capacity (ORAC) assay, ferric reducing antioxidant power (FRAP) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH?) and 2, 2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS?+) radical scavenging assays. Characterization and quantification of five polyphenol compounds which were previously identified to be present in all the selected plants were carried out. None of the extracts showed a decrease in the total phenolics content or the ORAC and FRAP values following digestion. None of the quantified phenolic compounds had decreased during any of the digestion phases – an observation which was deemed as beneficial in terms of therapeutic properties. Overall, the parameters analyzed were relatively stable throughout the digestive process in all the extracts.

     

Comparative Study of Lipophilic and Hydrophilic Antioxidants from <Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">In vitro</Emphasis> Grown <Emphasis Type="Italic">Coriandrum sativum</Emphasis>

Coriander is commonly used for medicinal purposes, food applications, cosmetics and perfumes. Herein, the production of antioxidants in vegetative parts (leaves and stems) of in vivo and in vitro grown samples was compared. In vitro samples were clone A- with notorious purple pigmentation in stems and leaves and clone B- green. Seeds were also studied as they are used to obtain in vivo and in vitro vegetative parts. Lipophilic (tocopherols, carotenoids and chlorophylls) and hydrophilic (sugars, ascorbic acid, phenolics, flavonols and anthocyanins) compounds were quantified. The antioxidant activity was evaluated by radical scavenging activity, reducing power and lipid peroxidation inhibition. The in vivo sample showed the highest antioxidant activity mainly due to its highest levels of hydrophilic compounds. Otherwise, in vitro samples, mainly clone A, gave the highest concentration in lipophilic compounds but a different profile when compared to the in vivo sample. Clones A and B revealed a lack of β-carotene, β- and δ-tocopherols, a decrease in α-tocopherol, and an increase in γ-tocopherol and clorophylls in comparison to the in vivo sample. In vitro culture might be useful to explore the plants potentialities for industrial applications, controlling environmental conditions to produce higher amounts of some bioactive products.     

Characterizing the pigmented traditional rice cultivars grown in temperate regions of Kashmir (India) for free and bound phenolics compounds and in vitro antioxidant properties

The purpose of the research was to identify the phenolic and flavonoid compounds of seven different traditional pigmented whole rice cultivars grown in the temperate regions of Kashmir so as to study their relationship with in vitro antioxidant capacities. The completely pigmented rice cultivars were found to have higher phenolic, flavonoid, anthocyanin contents and exhibited higher antioxidant capacities than the light colored and sparely colored rice cultivars. A total of 40 compounds had been identified in the analyzed rice cultivars that were found to be distributed in 6 major categories with 6-phenolics, 6-flavonoids, 11-hydroxycinnamic acid derivatives, 7-hydroxybenzoic acid derivatives, 3-anthocyanins and 7-flavonoid glucosides of different flavonoid compounds. Among the free and bound fractions for each cultivars the light and sparsely colored depicted higher content of phenolics and in vitro antioxidant properties in bound faction, while the completely pigmented cultivars showed higher antioxidant properties in free fractions. The anthocyanins quercetin-3-O-galactoside, cyanidin-3-O-rutinoside and pelargonidin-3-O-diglucoside had been identified by LC-MS existing in the free fractions of the analyzed rice cultivars whereas, the free fraction of acetone + H₂O possessed higher percentage of phenolic compounds as compared to methanolic extracts and bound fractions. The black colored cultivars possessed higher DPPH scavenging activity and lipid peroxidation inhibition.     

野葛愈伤组织提取物体外清除自由基活性的研究

为确定野葛愈伤组织提取物的清除自由基和抗氧化活性及开发新型天然抗氧化剂提供试验依据,采用1,1-二苯基苦基苯肼(DPPH.)的有机自由基体系以及羟自由基和超氧阴离子自由基两种无机活性氧自由基体系,检测野葛愈伤组织提取物的体外清除自由基和抗氧化活性,并与野葛根提取物、葛根素、Vc和茶多酚进行对比研究。结果表明:野葛愈伤组织提取物对DPPH.自由基的清除能力较强,与野葛根提取物、茶多酚的清除能力相当,且显著优于葛根素和Vc;野葛根提取物的羟自由基(.OH)清除活性最高,茶多酚和野葛愈伤异黄酮次之,Vc对.OH的清除活性稍弱;野葛愈伤组织提取物的羟自由基清除活性随处理浓度增加而提高;野葛愈伤组织提取物、野葛根提取物和Vc均具有较强的清除O-2.能力,均显著高于葛根素和茶多酚。说明野葛愈伤组织提取物具有较强的清除自由基和抗氧化活性。     

Phenolics,Sugars, Antimicrobial and Free-Radical-Scavenging Activities of <Emphasis Type="Italic">Melicoccus bijugatus</Emphasis> Jacq. Fruits from the Dominican Republic and Florida

Edible fruits of the native South American tree Melicoccus bijugatus Jacq. are consumed fresh or in traditional food, drink and medicinal preparations. Some therapeutic effects of these fruits may be due to phenolics and sugars. Aqueous acetone, methanol or ethanol tissue extracts of different cultivars or collections of M. bijugatus fruits from the Dominican Republic and Florida were analyzed for total phenolics and free radical scavenging activity by UV-vis spectroscopy, sugars by gas chromatography, and antimicrobial activity by the disc diffusion assay. Total phenolics and free radical scavenging activities ranked: seed coat > embryo > pulp extracts. Montgomery cultivar fruits had the highest total phenolics. For sugars: pulp > embryo and highest in Punta Cana fruit pulp. In all extracts: sucrose > glucose and fructose. Glucose:fructose ratios were 1:1 (pulp) and 0.2:1 (embryo). Pulp extracts had dose-response antibacterial activity and pulp and embryo extracts had antifungal activity against one yeast species. Phenolics and sugars were confirmed with thin-layer chromatography and nuclear magnetic resonance. Sugar-free pulp fractions containing phenolics had slightly more antimicrobial activity than H₂O-soluble pulp fractions with sugars. Results indicate M. bijugatus fruits contain phenolics, sugars and other H₂O-soluble compounds consistent with therapeutic uses. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Chemical and Functional Characterization of Kañiwa (C<Emphasis Type="Italic">henopodium pallidicaule</Emphasis>) Grain,Extrudate and Bran

Cereals provide a good source of dietary fibre and other important compounds with nutritional potential, such as phenolic compounds, antioxidants, minerals and vitamins. Although native Andean cereals are known to have high nutritional value, their minor components have not been studied thoroughly. In this study, two varieties of a native Andean crop, kañiwa (Chenopodium pallidicaule), were investigated as sources of dietary fibre and specific antioxidant compounds. Two products, an extrudate and bran, were also prepared and their functional properties and bioactive compounds were determined. Both varieties were rich in total dietary fibre and lignin, and the phenolic components analyzed had high antioxidant activity. The extrudates had good functional properties, such as degree of gelatinization, sectional expansion index and water solubility index; the bran was high in bioactive compounds, such as total phenolics. In conclusion, kañiwa may offer an alternative to traditional cereals as a health-promoting food ingredient.     

Color and Antioxidant Characteristics of Some Fresh Fig (Ficus carica L.) Genotypes from Northeastern Turkey

Fruit skin color, total phenolics, total anthocyanins, soluble solids content, titratable acidity and total antioxidant capacity in fresh fruits of a number of local and well-known fig (Ficus carica L.) genotypes and cultivars grown in northeastern Turkey were determined. TEAC (Trolox equivalent antioxidant capacity) and FRAP (ferric reducing antioxidant power) assays were used to determine total antioxidant capacity. Fruit skin color of genotypes were found to be very diverse, i.e., light green, light purple, purple, dark purple and black. The content of total phenolics, soluble solids content (SSC) and titratable acidity greatly varied in the range from 24 to 237?mg of gallic acid equivalent per 100?g fresh weight, 18.60 to 26.30?% and 0.16 to 0.47?% in local genotypes and studied cultivars. In general, total antioxidant capacities determined by two methods expressed higher values in the local fig genotypes compared with the cultivars. The results suggested that genotype is the main factor that determines difference in the composition of bioactive compounds in figs and provide information on putative health benefits locally grown genotypes.     

Polyphenolic Composition and Antioxidant Characteristics of Kumquat (<Emphasis Type="Italic">Fortunella margarita</Emphasis>) Peel Fractions

The polyphenolic composition of two Fortunella margarita (Nagami kumquat) specimens from Greece and Egypt was investigated employing fractionation by solvent partition and liquid chromatography-mass spectrometry. The main groups of phenolics identified in the different fractions generated were C-glycosylated flavones, O-glycosylated flavones, C-glycosylated flavanones, O-glycosylated flavanones, flavonols, chalcones, phenolic acids and derivatives thereof. The antioxidant potency of the fractions was assessed using two representative in vitro tests, including antiradical activity and hydroxyl free radical scavenging activity. It was revealed that the ethyl acetate fractions from both specimens contained the higher polyphenol content and exhibited the better antioxidant characteristics. The results indicated that F. margarita peels may be regarded as a rich source of potentially bioactive polyphenols.     

Antioxidant properties of wheat and rye bran extracts obtained by pressurized liquid extraction with different solvents

Rye and wheat brans are valuable sources of bioactive compounds, which could be used for the development and commercialization of high added value functional ingredients such as dietary antioxidants. The aim of this study was to evaluate antioxidant potential of rye and wheat bran using different polarity solvents. Cereal brans were ground to four different particle size fractions and extracted at 10.3 MPa pressure and 80 °C temperature by consecutive application of hexane, acetone and methanol:water (80:20%). The highest extract yield was obtained from rye bran using methanol-water; particle size in most cases had a significant effect. Antioxidant potential of extracts was assessed by ABTS^+•, DPPH^• scavenging, ORAC and total phenols content (TPC) assays. Extraction solvent had a major influence on TPC and antioxidant activity of the extracts. The most active extracts were obtained using methanol:water; rye bran extracts, in general, were stronger antioxidants than wheat bran extracts. For the majority of assays, reduction of particle size resulted in higher antioxidant activity values. However, ABTS^+• scavenging was found to decrease by decreasing particle size of rye bran used for extraction.     

Oat phenolic content and total antioxidant capacity during grain development

Oats (Avena sativa L.) were revaluated in recent years as a promising crop for improving the nutritional quality of foods, due to their richness in many bioactive compounds, including phenols. These plant secondary metabolites are useful as radical scavenging, and also possess positive biochemical effects against cardiovascular diseases, cancer growth and age-related diseases. Twenty oat cultivars were analyzed for their soluble phenol content (SPC, ranging 0.78–1.09 g_GAE/kg d.m.) and total antioxidant capacity (TAC, ranging 13.99–18.84 mmol TE/kg d.m.). In another experiment, the kinetics of SPC accumulation and TAC in the immature grains of five oat cultivars revealed a marked decrease of both parameters (−48.9% and −72.1%, respectively) from ten days after anthesis to maturity. These results could suggest a possible use of immature oat grains in human nutrition, as it was already proposed for other cereals.     

Antioxidant potential of sesame (Sesamum indicum) cake extract in stabilization of sunflower and soybean oils

Recently natural antioxidants have gained increased interest because natural food ingredients are safer than synthetic ones. Antioxidant activities and protective effects of sesame cake extract (SCE) in stabilizing sunflower oil (SFO) and soybean oil (SBO) were tested. Since different antioxidant compounds have different mechanisms of action, 2,2-azinobis (3-ethylbenzthiazoline sulfonate) (ABTS) radical scavenging activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity, and β-carotene/linoleic acid test system were used to assess the antioxidant efficacy of SCE. Total phenolic, flavonoid and flavonol contents in SCE were 1.94 (mg gallic acid equivalent (GAE) g⁻¹ dry weight (DW)), 0.88 (mg quercetain equivalent (QE) g⁻¹ DW), and 0.40 (mg QE g⁻¹ DW), respectively. Protective effects of SCE in stabilizing SFO and SBO were tested, compared to synthetic antioxidants, by measuring their peroxide values (PV), conjugated dienes (CD), conjugated trienes (CT) and p-anisidine value during accelerated storage. Results indicated that SCE exhibited stronger antioxidant activity in SFO and SBO than butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), while its antioxidant activity was less than that of tert-butyl hydroquinone (TBHQ).