Efficiency of enzymatic and other alternative clarification and fining treatments on turbidity and haze in cherry juice

Several alternative strategies were examined for improving conventional juice fining procedures for cherry juice clarification and fining in laboratory-scale experiments: Centrifugation of freshly pressed juice from 1000g to 35,000g induced decreased turbidity according to a steep, negative power function. Individual and interactive effects on turbidity and haze formation in precentrifuged and uncentrifuged cherry juice of treatments with pectinase, acid protease, bromelain, gallic acid, and gelatin-silica sol were investigated in a factorial experimental design with 32 different parameter combinations. Gelatin-silica sol consistently had the best effect on juice clarity. Centrifugation of cherry juice (10,000g for 15 min) prior to clarification treatment significantly improved juice clarity and diminished the rate of haze formation during cold storage of juice. Both treatment of precentrifuged cherry juice with Novozym 89L protease and co-addition of pectinase and gallic acid improved cherry juice clarity and diminished haze levels. None of the alternative treatments produced the unwieldy colloids notorious to gelatin-silica sol treatment. The data suggest that several alternative clarification strategies deserve further consideration in large-scale cherry juice processing. Precentrifugation of juice before clarification and fining is immediately recommended.     

草莓胡萝卜复合澄清汁后浑浊机理研究

为探讨草莓胡萝卜复合澄清汁的后浑浊机理，研究了化学试剂、贮藏温度和顶隙大小对草莓胡萝卜复合澄清汁的澄清度、褐变指数、黄烷醇类多酚含量、可溶性蛋白含量的影响。结果表明：贮藏温度越高，包装顶隙越大，氧气存留越多，随贮藏时间延长，果蔬汁浑浊形成越多，褐变程度越大，黄烷醇类多酚降解程度越大，可溶性蛋白损失越多；果蔬汁中添加尿素和十二烷基磺酸钠(SDS)，都能减少果蔬汁中浑浊的产生，抑制产品褐变，减缓黄烷醇类多酚的降解，减少可溶性蛋白的损失，说明氢键和疏水键都是和浑浊有关的化学键。蔗糖的添加对果蔬汁浑浊的形成没有太     

Protease-assisted clarification of black currant juice: synergy with other clarifying agents and effects on the phenol content

Conventional clarification with gelatin and silica sol removes a considerable amount of antioxidant phenolics from berry juices. This study examined the clarification and haze-diminishing effects of alternative clarification strategies on black currant juice including centrifugation and addition of acidic protease and pectinolytic enzyme preparations and gallic acid. Centrifugation of freshly pressed juice (10,000 g for 15 min) resulted in a approximately 95% reduction of immediate turbidity and had a decreasing effect on haze development in the juice during cold storage without significantly compromising the total phenols levels. The extent of clarification and haze diminishment varied after individual treatments with five different acidic proteases, but one of the protease preparations, Enzeco, derived from Aspergillus niger, consistently tended to perform best. The individual and interactive effects on juice turbidity, total phenols, and total anthocyanin contents of clarification treatments involving the use of two selected acid proteases (Enzeco and Novozyme 89L), a pectinase (Pectinex BE 3-L), and gallic acid were evaluated in a full factorial 2(4) experimental design. Haze development during cold storage decreased when gallic acid or any of the enzyme preparations were employed individually, but negative interaction effects resulted when the pectinase was employed in combination with any of the proteases. After 28 storage days at 2 degrees C, the lowest levels of haze formation were achieved when the Enzeco protease preparation, added at 0.025 g/L, was added with 0.050 g/L of gallic acid and allowed to react in the juice for 90 min at 50 degrees C. The corresponding anthocyanin reduction was approximately 12% (compared to approximately 30% with gelatin silica sol treatment). The data support the hypothesis that phenol-protein interactions are involved in juice turbidity development during cold storage of berry juices and demonstrate that precentrifugation and protease-assisted clarification show promise as an alternative, phenolics-retaining clarification strategy in black currant juice processing.     

Sulfate--a candidate for the missing essential factor that is required for the formation of protein haze in white wine

Protein haze formation in white wine is dependent on the presence of both wine protein and other unknown wine components, termed factor(s) X. The ability to reconstitute protein haze upon heating artificial model wine solutions (500 mg/L thaumatin, 12% ethanol, 4 g/L tartaric acid) to which candidate components were added was employed to identify factor(s) X. No protein haze was formed in the absence of additives. The individual or combined addition of caffeic acid, caftaric acid, epicatechin, epigallocatechin-O-gallate, gallic acid, or ferulic acid at typical white wine concentrations did not generate protein haze. However, PVPP fining of commercial wines resulted in a reduction in protein haze, suggesting that phenolic compounds may play a modulating role in haze formation. To elucidate the nature of the unknown factor(s) wine was fractionated and fractions were back-added to model wine and tested for their essentiality. Wine fractions were generated by ultrafiltration, reverse-phase chromatography, and mixed-mode anion-exchange and reverse-phase chromatography. The only purified fraction containing the essential component(s) was free of phenolic compounds, and analysis by mass spectrometry identified sulfate anion as the dominant component. Reconstitution with KHSO4 using either commercially available thaumatin or wine proteins confirmed the role of sulfate in wine protein haze formation. The two main wine proteins, thaumatin-like protein and chitinase, differed in their haze response in model wines containing sulfate. Other common wine anions, acetate, chloride, citrate, phosphate, and tartrate, and wine cations, Fe(2+/3+) and Cu(+/2+), when added at typical white wine concentrations were not found to be essential for protein haze formation.     

Characterization of haze-active proteins in apple juice

The nature of the haze-active protein (HAP) in apple juice was investigated. Heat treatment removed protein indiscriminately while polyvinylpolypyrrolidone (PVPP) treatment was fairly specific for proteins of 15 and 28 kDa. Presumably, the PVPP bound to polyphenols, which in turn were complexed with protein. Three candidate apple HAPs were isolated. Two were extracted from juice with acetone and fractionated by hydrophobic interaction chromatography and solid phase extraction with C18 (HAP I) or SAX (HAP II) material. Hydroxyproline-rich protein was extracted from apple tissue (HAP III). The order of haze formation with tannic acid was gliadin > HAP III > HAP II > HAP I > bovine serum albumin, which shows increasing haze formation with increasing proline content. The sizes of HAP I, II, and III were 28, 15, and 12 kDa; the first two corresponded to the sizes of proteins removed by PVPP treatment and are involved in juice haze formation.     

Effect of processing techniques at industrial scale on orange juice antioxidant and beneficial health compounds

Phenolic compounds, vitamin C (L-ascorbic acid and L-dehydroascorbic acid), and antioxidant capacity were evaluated in orange juices manufactured by different techniques. Five processes at industrial scale (squeezing, mild pasteurization, standard pasteurization, concentration, and freezing) used in commercial orange juice manufacturing were studied. In addition, domestic squeezing (a hand processing technique) was compared with commercial squeezing (an industrial FMC single-strength extraction) to evaluate their influences on health components of orange juice. Whole orange juice was divided into soluble and cloud fractions after centrifugation. Total and individual phenolics were analyzed in both fractions by HPLC. Commercial squeezing extracted 22% more phenolics than hand squeezing. The freezing process caused a dramatic decrease in phenolics, whereas the concentration process caused a mild precipitation of these compounds to the juice cloud. In pulp, pasteurization led to degradation of several phenolic compounds, that is, caffeic acid derivatives, vicenin 2 (apigenin 6,8-di-C-glucoside), and narirutin (5,7,4'-trihydroxyflavanone-7-rutinoside) with losses of 34.5, 30.7, and 28%, respectively. Regarding vitamin C, orange juice produced by commercial squeezing contained 25% more of this compound than domestic squeezing. Mild and standard pasteurization slightly increased the total vitamin C content as the contribution from the orange solids parts, whereas concentration and freezing did not show significant changes. The content of L-ascorbic acid provided 77-96% of the total antioxidant capacity of orange juice. Mild pasteurization, standard pasteurization, concentration, and freezing did not affect the total antioxidant capacity of juice, but they did, however, in pulp, where it was reduced by 47%.     

Phenolic antioxidants and antiatherogenic effects of Marula (Sclerocarrya birrea Subsp. caffra) fruit juice in healthy humans

Antioxidant activity and composition of Israeli-grown marula ( Sclerocarrya birrea subsp. caffra) fruit juice and health-promoting aspects of juice consumption on serum lipids and lipoproteins pattern in healthy volunteers were studied. Marula juice was found to contain high vitamin C and potassium levels and low sugar concentration (267 mg dL (-1), 328 mg dL (-1), and 7.3 g dL (-1), respectively). The juice contains a significant level of phenolics (56 mg of pyrogallol equiv dL (-1)) and was found to be a potent antioxidant (382 mg of vitamin C equiv dL (-1)). The antioxidant activity was resistant to pasteurization regimens and long-term freezing and slowly decreased during refrigeration, losing up to 14% of its capacity after 4 weeks. Three-week administration of the juice as a food supplement to healthy subjects significantly reduced their serum total cholesterol (by 8%), LDL-cholesterol concentration (by 17%), and triglyceride level (by 7%), increased their serum HDL-cholesterol level (by 10%), and attenuated serum oxidative stress. Upon a 4 week "washout" period, most of these parameters returned toward baseline values. Separation of the juice soluble phenolics by HPLC produced potent antioxidant fractions, tentatively containing hydrolyzable tannins, catechins, and hydroxycinnamic acid derivatives, which could be responsible for the observed protection against atherosclerosis risk factors following marula fruit juice consumption.     

Processing strawberries to different products alters contents of vitamin C, total phenolics, total anthocyanins, and antioxidant capacity

Strawberries were processed to juice, nectar, wine, and puree. For investigation of the antioxidant capacity as well as the contents of ascorbic acid, total phenolics and total anthocyanins, samples were taken after different stages of production to determine the effects of processing. The content of vitamin C was measured spectrophotometrically. The total phenolic content was analyzed by using the Folin-Ciocalteu method, and the amount of total anthocyanins was determined by using the pH-differential method. Two different methods-the trolox equivalent antioxidant capacity assay and the ferric reducing antioxidant power test-were used to determine the hydrophilic antioxidant capacity. This study showed the decrease of all investigated parameters within processing strawberries to different products. The content of ascorbic acid decreased with production time and processing steps, especially during heat treatment. The investigations on total phenolics in strawberry products proved fining to be a mild method to clarify berry juices and wines without removing high amounts of total phenolics. Fermentation did not lead to heavy losses of total phenolics, probably due to polymerization and condensation of monomer phenolics such as anthocyanins. Total anthocyanins and the hydrophilic antioxidant capacity decreased while using high temperatures. Anthocyanins also decreased considerably during the processing of wines, mainly caused by fermentation and pasteurization.     

Chemical composition of clarified bayberry (Myrica rubra Sieb. et Zucc.) juice sediment

Clarified bayberry juice turned hazy upon storage at 25 degrees C for 6 months, and the chemical composition of centrifugally separated sediment was analyzed. Bayberry juice haze was mainly protein-tannin haze. The lyophilized sediment contained 20.4 +/- 4.3% of protein, 70.2 +/- 2.6% of total polyphenols, 7.2% of monosaccharides, and 6.7 +/- 0.6% of ash. Amino acid analyses and molecular weight distribution estimation indicated that bayberry proteins were haze-active proteins with a molecular weight less than 8 kDa. Gallic acid, quercetin hexoside, quercetin deoxyhexoside, and quercetin were found in the methanol-dissolved sample, while gallic acid, protocatechuic acid, cyanidin, ellagic acid, and quercetin were detected in the acid-hydrolyzed sample. Ellagic acid was the dominant individual phenolic (9.9 +/- 0.19 g/100 g dry weight, 55.3% of the total amount) in the sediment. Monosaccharides of rhamnose, arabinose, mannose, glucose, and galactose in the sediment were most probably the glycoside moieties of the anthocyanins, flavonols, and elllagitannins. Metal ions of calcium, magnesium, potassium, iron, and copper also indicated the heterogeneous characteristics of the sediment.     

Contribution of tomato phenolics to antioxidation and down-regulation of blood lipids

This study was performed to understand the characteristics and biological activities of phenolics in tomatoes and to examine the effect of tomato on the regulation of blood lipids. Tomatoes of both big and small sizes were used fresh, after blanching, or after blanching and heating. Moreover, a human clinical trial was conducted to examine plasma antioxidation, status of blood lipids, and phenolic responses after ingestion of fresh tomato, tomato juice, and a lycopene drink. The contents of tomato phenolics were increased by 34% for small tomato and by 23% for big tomato after treatment by blanching and heating at 100 degrees C for 30 min. Tomato phenolics showed fair antioxidant activity (57-71%) and also synergistically promoted the antioxidation (81-100%) of tomato carotenoids. In the human clinical study, total antioxidant capacity and phenolic contents in plasma were increased after administration of fresh tomato and tomato juice, but no significant difference was found for lycopene drink consumption. Triglyceride levels and low-density lipoprotein cholesterol were decreased after administration of fresh tomato and tomato juice, and high-density lipoprotein cholesterol was increased.     

Stable isotopic carbon composition of apples and their subfractions--juice, seeds, sugars, and nonvolatile acids

The 13C:12C ratios of 8 authentic apple juice samples and their subfractions were determined by mass spectrometry. Apples from Argentina, Mexico, New Zealand, and the United States were processed into juice; pulp was collected from the milled fruit and seeds were collected from the press-cake. Sugars, nonvolatile acids, and phenolics were isolated from the juice by treatment with ion-exchange resins and polyvinylpyrrolidone (PVPP). The mean value for all juice samples was -24.2% which is close to the values reported by other investigators. Juice from apples grown in Argentina, Mexico, and New Zealand did not differ from U.S. samples. The isotopic composition of the subfractions ranged from -22.0 to -31.0%. The values for the pulp were essentially the same as for juice. The sugar fraction was slightly less negative than the juice; the nonvolatile acid and phenolic fractions were more negative. The levels of nonvolatile acids and phenolics in apple juice are low, however, so these compounds contribute little to overall delta 13C values in juice.     

Simple and rapid method for the analysis of phenolic compounds in beverages and grains

A new method for the detection of phenolics in food systems was developed. This method is based on interactions of phenolics with Fast Blue BB diazonium salt in alkali pH, forming azo complexes, with the absorbance measured at 420 nm after 60 min. The linear regression correlations (R(2)) of gallic acid calibration standards were >0.99. The phenolic content (gallic acid equivalent) of samples analyzed yielded higher ratios (1.7-6.6) of the total phenolics by Fast Blue BB to Folin-Ciocalteu methods in most beverages and grain samples, but in flaxseed and some juice blends, the ratios were <1. The lower ratios suggest the presence of non-phenolic reducing constituents measured with the Folin-Ciocalteu method as "total phenolics". This method is simple and inexpensive and can be used to rapidly assess the total phenolics of foods and beverages.     

Antioxidant and antiproliferative activities of raspberries

Raspberries are rich in phenolic phytochemicals. To study the health benefits of raspberries, four fresh raspberry varieties (Heritage, Kiwigold, Goldie, and Anne) were evaluated for total antioxidant and antiproliferative activities. The total amount of phenolics and flavonoids for each of the four raspberry varieties was determined. The Heritage raspberry variety had the highest total phenolic content (512.7 +/- 4.7 mg/100 g of raspberry) of the varieties measured followed by Kiwigold (451.1 +/- 4.5 mg/100 g of raspberry), Goldie (427.5 +/- 7.5 mg/100 g of raspberry), and Anne (359.2 +/- 3.4 mg/100 g of raspberry). Similarly, the Heritage raspberry variety contained the highest total flavonoids (103.4 +/- 2.0 mg/100 g of raspberry) of the varieties tested, followed by Kiwigold (87.3 +/- 1.8 mg/100 g of raspberry), Goldie (84.2 +/- 1.8 mg/100 g of raspberry), and Anne (63.5 +/- 0.7 mg/100 g of raspberry). The color of the raspberry juice correlated well to the total phenolic, flavonoid, and anthocyanin contents of the raspberry. Heritage had the highest a/b ratio and the darkest colored juice, and the Anne variety showed the lowest phytochemical content and the palest color. Heritage raspberry variety had the highest total antioxidant activity, followed by Kiwigold and Goldie, and the Anne raspberry variety had the lowest antioxidant activity of the varieties tested. The proliferation of HepG(2) human liver cancer cells was significantly inhibited in a dose-dependent manner after exposure to the raspberry extracts. The extract equivalent to 50 mg of Goldie, Heritage, and Kiwigold fruit inhibited the proliferation of those cells by 89.4 +/- 0.1, 88 +/- 0.2, and 87.6 +/- 1.0%, respectively. Anne had the lowest antiproliferative activity of the varieties measured but still exhibited a significant inhibition of 70.3+/- 1.2% with an extract equivalent to 50 mg of fruit. The antioxidant activity of the raspberry was directly related to the total amount of phenolics and flavonoids found in the raspberry (p < 0.01). No relationship was found between antiproliferative activity and the total amount of phenolics/flavonoids found in the same raspberry (p > 0.05).     

Accumulation of bioactive compounds during growth and development of mango ginger (Curcuma amada Roxb.) rhizomes

Accumulation of bioactive compounds and storage components during developmental stages of mango ginger ( Curcuma amada Roxb.) rhizome was investigated from 60 to 240 days, as a function of physiological maturity. Four distinct developmental phases were defined, namely, vegetative phase (up to 60 days from planting), initiation and development phase (60-150 days), maturation phase (150-180 days), and senescence phase (180 days). Difurocumenonol, a bioactive terpenoid compound and phenolics were identified as biomarkers, to determine the optimum physiological maturity to harvest mango ginger rhizome. Accumulation of phenolics was observed in newly initiated rhizomes (after 60 days from planting). The phenolic content was high in mango ginger pulp compared to its juice. Newly initiated rhizome contained no difurocumenonol, and it was observed after 120 days after planting. Peak accumulation of phenolics, difurocumenonol, and total protein were noticed in 180 day old rhizome. Accordingly, the abundance of these components on 180 days was set as an optimum maturity standard for harvest of mango ginger rhizome, compared with a conventional harvest period that ranges from 200 to 240 days.     

Determination of the grape invertase content (using PTA-ELISA) following various fining treatments versus changes in the total protein content of wine. relationships with wine foamability

Proteins have proven to play a major role in the stabilization of foam in Champagne wines despite their low concentration that ranges from 4 to 20 mg/L. The aim of this study was to evaluate the effect of fining on total protein and grape invertase contents of champenois base wines and their foaming properties. Data showed that fining and especially the use of bentonite at doses ranging from 10 to 50 g/hL leads to a significant decrease in the total protein content of wines together with that of the grape invertase content, with such a decrease being very detrimental to the foaming properties of the treated wines in terms of foam height (HM) and foam stability (HS). Only a slight decrease in the total protein content, in the grape invertase concentration, and in the foam quality of wines was observed when using casein (10 and 20 g/hL) or bentonite combined with casein (both at 20 g/hL). Our study thus clearly establishes the good correlation existing between the wine protein concentration and its foaming properties. A remarkable correlation was observed between the decrease in the grape invertase content and the total protein content of wines, following bentonite treatments, suggesting that the grape invertase (which represents at least 10-20% of the wine proteins) follows a similar behavior upon fining to other proteins of Champagne wines, despite the high molecular mass and the highly glycosylated structure of this particular protein. Moreover, the decrease in total protein and grape invertase contents of wine after fining with bentonite was found to be correlated with a decrease in the foaming properties of the corresponding wines (with respectively R(2) = 0.89 and 0.95).     

Haze formation in model beer systems

The interaction of a haze-active protein (gliadin) and a haze-active polyphenol (tannic acid) was studied in a model beer system in order to investigate the principle mechanisms of haze formation at low temperatures. Low concentrations (g/L) of tannic acid, high concentrations of gliadin, and comparatively high temperatures lead to maximum haze values. When considered on a molar basis, the greatest haze levels are displayed at an approximate 1:1 equivalence of polyphenol and protein. The greater part of haze formation was completed within 0.5 h, irrespective of the concentration of gliadin, the concentration of tannic acid, and the temperature of the model solution.     

Clarification of Muscat musts using wheat proteins and the flotation technique

The bovine spongiform encephalopathy (BSE, mad cow) crisis has led some wine-makers to question gelatin as a fining agent and to reject the use of these animal proteins. The search for a substitute for gelatin was begun by comparing vegetable proteins (particularly gluten) with gelatin fining treatments. Clairette de Die (French-controlled appellation) is a sparkling sweet wine. The alcoholic fermentation begins in a tank, continues in a crown-cap bottle, and stops before the complete consumption of sugar. All particles present in the bottle have to be removed before corking, and it is important to have a must as clear as possible. This study concerned the clarifying of a Muscat must, treated with pectinases, using a very efficient flotation technique. The must is fined with bentonite/silica/protein (fish gelatin, wheat gluten, or lupin isolate) to induce flocculation and then pressurized (6 bar). After depressurization, microbubbles cling to flocculates and climb to the top of the flotation tank (this flotation foam is clarified using a rotary filter). At laboratory scale, gluten (20 g/hL) and gelatin (10 g/hL) (each combined with bentonite and silica gel) gave turbidities of 50 and 35 NTU, respectively (6.5 and 4.1% of that of the nontreated must). The Muscat must was also clarified by static settling. The turbidity decreased by 86% for the gluten/bentonite fining and by 60% for the gelatin/bentonite fining. Visually, gluten flocculation takes longer to occur and flocculates sedimentation is longer than with gelati, but the removal of insoluble particles is more complete and leads to lower turbidities. At an industrial scale, gluten (20 g/hL) and gelatin (10 g/hL) (each combined with bentonite and silica gel) gave turbidities of 60 and 48 NTU, respectively. Turbidities measured in the tanks 14 h after the flotation showed a better efficiency for the wheat gluten (24 NTU) compared to gelatin (28 NTU). This is explained by the static settling that completed the clarifying effect of the flotation. The last experiment showed comparable efficiencies for wheat gluten and lupin proteins. BSE caused a situation of crisis (in Europe particularly) leading the public and wine-makers to lose their confidence in the use of gelatin as fining agent and to reject animal proteins in general. It is proposed here that vegetable proteins could efficiently replace gelatin.     

Molecular properties and activities of tuber proteins from starch potato cv. Kuras

Potato starch production leaves behind a huge amount of juice. This juice is rich in protein, which might be exploited for food, biotechnological, and pharmaceutical applications. In northern Europe cv. Kuras is dominant for industrial starch production, and juice protein of freshly harvested mature tubers was fractionated by Superdex 200 gel filtration. The fractions were subjected to selected activity assays (patatin, peroxidase, glyoxalases I and II, alpha-mannosidase, inhibition of trypsin, Fusarium protease, and alcalase) and protein subunit size determination by SDS-PAGE and mass spectrometry. Proteins present in SDS-PAGE bands were identified by tryptic peptide mass fingerprinting. Protein complexes such as ribosomes and proteasomes eluted with the void volume of the gel filtration. Large proteins were enzymes of starch synthesis dominated by starch phosphorylase L-1 (ca. 4% of total protein). Five identified dimeric patatin variants (25%) coeluted with four monomeric lipoxygenase variants (10%) at 97 kDa. Protease inhibitor I variants (4%) at 46 kDa (hexamer) inhibited alcalase. Fourteen Kunitz protease inhibitor variants (30%) at 19 kDa inhibited trypsin and Fusarium protease. Carboxypeptidase inhibitor variants (5%) and defensins (5%) coeluted with phenolics. The native sizes and molecular properties were determined for 43 different potato tuber proteins, several for the first time.     

微波预处理原料对苹果汁褐变的影响

为了防止苹果汁在贮藏过程中褐变,采用不同的微波功率和时间预处理原料苹果,研究微波预处理对苹果汁褐变的影响。结果表明,微波处理可以提高苹果汁的色值,降低多酚氧化酶的活性,引起氨基态氮含量的下降,并使苹果汁的酸度略微提高。在微波功率为720～900 W,处理时间75～125 s时,果汁色值较高。室温贮存45 d后,微波功率900 W,时间为100 s处理的果块,果汁色值为67.8,比未经微波处理的高91.5%。微波预处理是防止苹果汁褐变的一种简便安全、合理经济的加工方法。     

Extraction of anthocyanins and other phenolics from black currants with sulfured water

Health benefits of fruits, vegetables, and red wine are attributed to anthocyanins and other phytochemicals. In this research, the extraction of phenolics from black currants was optimized using different SO(2) concentrations (28, 300, 700, 1100, and 1372 ppm), temperatures (6, 20, 40, 60, and 74 degrees C), and solvent to solid ratios (S/S) (6, 20, 40, 60, and 74 mL/g). Surface response methodology was used to optimize yields of anthocyanins and total phenolics, as well as their antiradical and antioxidant activities. The extraction of phenolics varied with the SO(2) concentration, S/S, and temperature. Maximum yields of total phenolics and anthocyanins were obtained at an SO(2) concentration of 1000-1200 ppm and 19 L of solvent/kg of milled frozen berries. The increase of extraction temperature increased the rate of extraction and, thus, times to reach equilibrium for the extraction of total phenolics and anthocyanins were reduced. However, for the extraction of anthocyanins it is recommended that temperatures of 30-35 degrees C be used, as higher temperatures will degrade these compounds. Antioxidant activity was affected by all three experimental variables evaluated; however, the main variable affecting it was S/S. The higher the S/S, the lower the antioxidant index.