Effects of phenolic compounds in blueberries and muscadine grapes on HepG2 cell viability and apoptosis

Although blueberries and muscadine grapes have high contents of polyphenols, few studies have been conducted to assess their potential effects on cancer cells. The objective of this study was to systematically evaluate the effects of different fractions of phenolic compounds in blueberries and muscadine grapes on HepG2 liver cancer cell viability and apoptosis. Three cultivars of blueberries (‘Briteblue’, ‘Tifblue’ and ‘Powderblue’) and four cultivars of muscadine grapes (‘Carlos’, ‘Ison’, ‘Noble’, and ‘Supreme’) were assessed in this study. Polyphenols were extracted and further separated into phenolic acids, tannins, flavonols, and anthocyanins using a HLB cartridge and LH-20 column. The major compounds of different fractions were characterized. The phenolic acid fractions of muscadine grapes and blueberries showed a 50% inhibition of HepG2 cell population growth at the level of 1–2 mg/mL. The greatest inhibitory effects were observed from the anthocyanin fractions with 50% inhibitions of cancer cell population growth at concentrations of 70–150 and 100–300 μg/mL in blueberries and muscadine grapes, respectively. The flavonol and tannin fractions showed intermediate activities. In addition, DNA fragmentation was measured by using a Cell Death Detection ELISA kit to assess the induction of apoptosis. The anthocyanin fraction resulted in a two- to fourfold increase in DNA fragmentation compared to control in both muscadine grapes and blueberries. These findings of inhibition of cancer cell growth and induction of apoptosis suggest that blueberries and muscadine grapes may contribute to reduction in liver cancer risk.     

圆叶葡萄种子发育过程中多酚积累特性

以栽培于广西地区2 个圆叶葡萄品种‘Noble’和‘Carlos’为实验材料,以真葡萄品种‘赤霞珠’和‘雷司令’为对照,使用超高效液相色谱-串联三重四极杆飞行时间质谱,分析其种子发育过程中多酚组成和积累特性。结果表明：在4种葡萄发育过程的种子样品中共检测到106种多酚,包括48种鞣花酸及前体物、24种黄酮醇、20种羟基苯甲酸、9种黄烷-3-醇、3种芪类和2种羟基肉桂酸。鞣花酸及前体物（主要是前体物）和黄烷-3-醇（主要是没食子酰化体和单体）分别是圆叶葡萄和真葡萄种子中最主要的多酚类型,但其含量在不同年份和品种间具有较大差异。前体物含量在圆叶葡萄‘Noble’幼果期种子中较高,然后逐渐下降,完全转色后再次上升;‘Carlos’则在浆果转色前种子中含量较高,随后略有下降。黄烷-3-醇在真葡萄‘雷司令’种子中2013年随着浆果发育含量显著升高,转色时含量达到最高,随后有所下降,2014年不受发育期的影响;‘赤霞珠’则分别在2013年幼果期和2014年果实完熟时的种子中含量最高。     

Changes in Free Amino Acid and Total Nitrogen Concentrations During Maturation of Muscadine Grapes (V. rotundifolia)

Free amino acids, ammonia and total nitrogen were followed during maturation of berries of Carlos and Noble muscadine grapes (V. rotundifolia). Chemical analysis was done on deseeded berries. Ammonia decreased in the berries of both cultivars during maturation. Total nitrogen content fluctuated and was not significantly correlated with maturity in either cultivar. Threonine (THR) and histidine + gamma aminobutyric acid (HIS + GABA) were the predominant free amino acids in both cultivars at an immature berry stage. At full berry maturity, arginine (ARG) was the predominant amino acid in Carlos, having increased over 100-fold during the sampling period while ARG, alanine (ALA), THR, and HIS + GABA accounted for 72.3% of the total free amino acids. ALA was the predominant free amino acid in mature berries of Noble and accounted for 30.4% of the total free amino acids. Of the additional cultivars sampled only at their normal harvest date, berries of Pride, Magnolia and Regale contained nearly equal amounts of ARG and ALA as the predominant amino acids. Mature Dixie berries had twice as much ALA (29.8%) as ARG (12.8%). In general, the free amino acids in berries of muscadine grapes were found in concentrations similar to those reported in the literature for V. vinifera and V. labrusca cultivars. However, the large increase in proline concentration reported to occur during maturation of berries of V. vinifera cultivars was not found in any of the muscadine cultivars examined.     

Muscadine Grape Polyphenoloxidase: Partial Purification by High Pressure Liquid Chromatography and Some Properties

A rapid method of isolating grape polyphenoloxidase (PPO) was developed, using two muscadine cultivars, “Noble” and “Welder”. PPO extracts from acetone powders were partially purified followed by filtration through 0.2μ membrane filter by HPLC. Reactivity of the active fraction per unit protein, increased 10-fold for Noble and 33-fold for Welder cultivars over values for the corresponding crude extracts. Electrophoretic analysis and activity stain showed that the enzyme displayed a single band in Welder and two bands for Noble. Optimum pH for PPO activity in Welder was pH 4.5 while that for Noble was pH 4.5–5.5. The enzyme in the Noble cultivar was relatively more active at alkaline pH. Activites of the enzymes were stable at temperatures up to 30°C for both cultivars.     

Influence of pressing method on juice stilbene content in muscadine and bunch grapes

ABSTRACT:  Juice stilbene contents of 2 muscadine ( Vitis rotundifolia) cultivars, Noble and Carlos, and 2 bunch grape ( Vitis labrusca) cultivars, Mid South and Miss Blanc were evaluated after crushing and pressing. Four commercial pressing methods were used to produce juice: (1) cold press of fresh berries (CP); (2) hot press of fresh berries (HP); (3) cold press of frozen and thawed berries (FCP); and (4) cold press of pectolytic enzyme treated berries (ECP). Free run juice (FRJ) from the crusher without pressing was also tested. An HPLC with a UV detector was used to verify the presence and quantity of resveratrol, piceid, and their isomers. For the muscadine juices, total stilbenes were greater for HP and FCP than for FRJ and CP treatments. Trans -resveratrol was only detected in HP juices of the muscadines at levels near the detection limits. The bunch grape cultivars behaved similar to the muscadines in the processing treatments. HP juice had the greatest total stilbene levels and was significantly greater than FRJ, CP, and FCP. Total stilbene levels in juices from the 2 bunch grape cultivars, were higher (up to 5 times greater depending on pressing method) than for the muscadine grape cultivars. Total stilbene levels in muscadine juices treated with pectinase before pressing (ECP) were similar to those of the FCP samples. Juice from FCP and ECP treated Noble muscadine had total stilbene levels greater than FRJ and CP but less than HP levels. Juices from HP-, ECP-, and FCP-treated Carlos muscadine had total stilbene levels greater than for FRJ and CP juices.     

Fatty acid composition,oxidative stability,antioxidant and antiproliferative properties of selected cold-pressed grape seed oils and flours

Cold-pressed chardonnay, muscadine, ruby red, and concord grape seed oils and their defatted flours were studied for their fatty acid composition, oxidative stability and antioxidant and antiproliferative activities. The phenolic profiles of the seed flours were also measured. The most abundant fatty acid in the oils was linoleic acid, ranging from 66.0 g/100 g of total fatty acids in ruby red seed oil to 75.3 g/100 g of total fatty acids in concord seed oil. The oils were also high in oleic acid and low in saturated fat. Ruby red grape seed oil recorded the highest oxidative stability index of 40 h under the accelerated conditions. Total phenolic content (TPC) was up to 100 times lower in the oils than in the flours. Lutein, zeaxanthin, cryptoxanthin, β-carotene, and α-tocopherol levels were also measured. DPPH radical-scavenging capacity ranged from 0.07 to 2.22 mmol trolox equivalents (TE)/g of oil and 11.8 to 15.0 mmol TE/g of flour. Oxidative stability of menhaden fish oil containing extracts of the seed flours was extended by up to 137%. HPLC analysis was conducted to determine the levels of free soluble, soluble conjugated and insoluble bound phenolics in the seed flours. The phenolic compounds analyzed included catechin, epicatechin, epicatechin gallate, quercetin, gallic acid, and procyanidins B1 and B2. Antiproliferative activity was tested against HT-29 colon cancer cells. All of the seed flours and muscadine seed oil registered significant (P < 0.05) inhibition of cancer cell growth. The results from this study demonstrate the potential of developing value-added uses for these seed oils and flours as dietary sources of natural antioxidants and antiproliferative agents for optimal health.     

高寒荒漠区延后栽培葡萄品质分析

为评价鲜食葡萄在高寒荒漠区延后栽培条件下的品质表现,以适应性较好的8个品种为研究对象,测定葡萄感官品质、内在品质、功效成分和糖酸品质,采用主成分分析法对果实品质进行综合分析和评价。结果表明,‘红地球’和‘紫地球’单果质量10 g以上,果粒大,能够保持原有的大果特征;无核品种较有核品种果粒小;8个品种果实果形指数均> 1,呈长椭圆形,除绿色品种外,果实着色较深;果皮颜色越深的品种,果皮花青素、类黄酮、总酚含量越高。2个无核品种(‘红宝石无核’和‘紫甜无核’)和‘红地球’可溶性固形物含量较高,在21%以上;糖酸组分及含量方面,果实中的糖分主要是果糖和葡萄糖,蔗糖未检出;酒石酸和苹果酸是主要有机酸,柠檬酸和草酸含量极微。酸甜风味方面,‘红宝石无核’口感最甜,其次为‘红地球’,‘瑞都香玉’和‘紫地球’口感较淡,‘摩尔多瓦’口感偏酸。通过主成分分析法提取了5个独立的主成分,累计贡献率87. 396%,以主成分为评价指标,对浆果品质进行综合评价,‘红宝石无核’、‘红地球’和‘玫瑰香’果实综合品质最好。     

The Proteins of Muscadine Grapes

The concentrations, molecular weights and isoelectric points of proteins present in muscadine grape cultivars were determined. The total protein contents of these grapes, are consistent with values reported in literature for V. vinifera grape proteins. Molecular weights of most of the protein fractions were, however, relatively high when compared with those of other grape cultivars. Isoelectric’ points of muscadine proteins ranged between pH 5.6 and pH 7.6.     

Antimicrobial effect of water-soluble muscadine seed extracts on Escherichia coli O157:H7

Water-soluble extracts were prepared from purple (cultivar Ison) and bronze (cultivar Carlos) muscadine seeds with or without heating. The Ison extracts had strong antimicrobial activity against a cocktail of three strains of Escherichia coli O157: H7. This extract had higher acidity (pH 3.39 to 3.43), total phenolics (2.21 to 3.49 mg/ml), tartaric acid (5.6 to 10.7 mg/ml), tannic acid (5.7 to 8.1 mg/ml), and gallic acid (0.33 to 0.59 mg/ml) than did the Carlos extracts. Heat treatment on both extracts increased antimicrobial activity, possibly because of increased acidity, tartaric acid, total phenolics, and individual phenolics. Heating of Ison extracts increased ellagic acid up to 83%. Up to 10.7 mg/ml tartaric acid alone was not as effective against E. coli O157:H7 as were water-soluble seed extracts. This finding suggests the involvement of other factors, such as tannic and gallic acids, in inactivation of this pathogen. Water-soluble muscadine seed extracts may be useful for incorporation into juice and other beverage products as natural preservatives.     

不同品种红麻籽油脂肪酸组成的GC-MS分析

分析9个不同品种红麻籽油脂肪酸组成及相对含量的差异。采用索氏提取法提取红麻籽油,并对其进行甲酯化处理后采用气相色谱-质谱联用(GC-MS)技术分析和鉴定红麻籽油脂肪酸组成及相对含量。结果表明:红麻籽含油率在19.63%~22.06%之间;红麻籽油的主要脂肪酸以亚油酸、油酸、软脂酸和硬脂酸为主,其中亚油酸甲酯(Z,Z)含量在31.98%~38.91%,油酸甲酯(E)含量在31.30%~36.93%,软脂酸甲酯含量在19.81%~23.02%,硬脂酸甲酯含量在3.32%~4.22%。因此,红麻籽油是一种典型的油酸-亚油酸型油脂。     

The fatty acid and tocopherol constituents of the seed oil extracted from 21 grape varieties (Vitis spp.)

BACKGROUND: Fatty acids and tocopherols in appropriate quantities are invaluable attributes that are desirable in seeds of agricultural products. Studies have generally focused on the evaluation of the oil and tocopherol components of oil crops. Recently, investigations revealed that the grape seed has robust potential in the production of healthy fatty acids as well as tocopherols. This study was thus conducted to determine the oil and tocopherol components of grape seeds, obtained from various grape cultivars of different species, including two rootstock varieties. RESULTS: The grape seed oil concentration of the studied varieties ranged from 7.3 to 22.4%. The determined fatty acid profiles of the genotypes conformed to the pattern described in the literature for grapes. Linoleic acid is the major component comprising 53.6–69.6% of the total, followed by oleic (16.2–31.2%), palmitic (6.9–12.9%) and stearic (1.44–4.69%). The oils of all the seeds analysed showed a preponderance of α‐tocopherol (ranging from 260.5 to 153.1 mg kg^?1 oil extract). β‐Tocopherol, γ‐tocopherol and δ‐tocopherol were also detected with the general means of 0.98, 22.2 and 0.92 mg kg^?1, respectively. Linoleic acid showed a significantly negative correlation with all the fatty acids analysed. The strongest negative correlation existed between linoleic and oleic acids (r = ? 0.834, P < 0.01). CONCLUSION: Present investigations indicated that oil content, fatty acid composition and tocopherol constituents of grape seed show great variation among the genotypes. Markedly higher proportions of linoleic acid with considerable amounts of tocopherols found in the oil samples suggest that grape seed is a good source for culinary, pharmaceutical and cosmetic uses. Copyright © 2012 Society of Chemical Industry     

Changes in Concentration of Certain Elements During Maturation of Muscadine Grapes (Vitis rotundifolia)

Changes in percent ash and certain elements were followed during maturation of berries of two cultivars of muscadine grapes (Vitis rotundifolia). Analyses were done on deseeded berries. Percent ash increased significantly during maturation of Carlos berries, but did not increase significantly during maturation of Noble berries. Potassium was the element in highest concentration in both cultivars during maturation and at harvest. Potassium, calcium, phosphorus, and sodium were the major elements found to be significantly correlated with maturation and were found in concentrations similar to those reported in the literature for V. vinifera grapes. Iron, copper, zinc, and manganese were minor components. Four additional muscadine cultivars, sampled only at full maturity, had similar concentrations of elements to those found in ripe berries of Carlos and Noble. The averaged values of ash (%) and elements (ppm) for all six cultivars when sampled at their normal harvest dates are: ash = 0.57; Na = 58.6 (Carlos and Noble only); P = 99.6; K = 1641.0; Ca = 124.2; Mg = 59.2; Mn = 7.2; Cu = 0.5; Zn = 0.5; Fe = 3.0.     

Fatty acids and triacylglycerols in the seed oils of Saudi Arabian date (Phoenix dactylifera L.) palms

Total lipids, fatty acids and triacylglycerols in seeds of the four Saudi Arabian date cultivars ‘Anbra’, ‘Megadwel’, ‘Sacai’ and ‘Sfwai’ were analysed. Total lipid contents ranged between 7.3 and 8.6%. Oleic, lauric and myristic acid represented the most abundant fatty acids in date seed oil. HPLC-DAD-ESI-MSⁿ analysis enabled the identification of 40 triacylglycerols with equivalent carbon number (ECN) 36–54, of which 17 were detected for the first time. Triacylglycerols mainly comprised LaOL, LaLaL, LaML, MML/LaPL and LaOO. ‘Anbra’ oil was characterised by highest proportions of saturated fatty acids and triacylglycerols with ECN 36 and 48–54. Unsaturated fatty acids prevailed in the three remaining varieties. Elevated proportions of triacylglycerols with medium ECNs were found in ‘Megadwel’ and ‘Sfwai’ oils. Aside from highest total lipid contents, ‘Sacai’ seeds predominantly contained triacylglycerols with low ECNs. Multivariate statistical analyses elucidated varietal influences on the fatty acid and triacylglycerol composition of date seed oil.     

Identification of anthocyanins in muscadine grapes with HPLC-ESI-MS

Muscadine grapes are important fruits grown in the southeastern United States. They are a good source of anthocyanins, but confirmatory analyses of the structure of individual anthocyanins in muscadine grapes using more advanced instrument are lacking. In this study, the anthocyanins of muscadine grapes were analyzed using HPLC-MS with electrospray ionization. Approximately 90% of the total anthocyanins were 3,5-diglucoside of delphinidin, cyanidin and petunidin; the remaining 10% were 3,5-diglucoside of peonidin and malvidin. There was a significant difference of total anthocyanin content among different cultivars of muscadine grapes with purple-skinned muscadine grapes having significantly higher levels of anthocyanins than bronze-skinned muscadine grapes. The total anthocyanin content in purple-skinned Jumbo and Cowart muscadine was 4.1 and 2.6 mg/g dry weight, respectively, while the total anthocyanin content in bronze-skinned Higgin and Carlos was negligible. The anthocyanins were mainly concentrated in the skin of the grapes. The results of the present study confirmed some of the anthocyanins reported by previous studies and also found one anthocyanin that might be falsely identified by previous studies.     

Effect of cold press and soxhlet extraction systems on fatty acid,tocopherol contents,and phenolic compounds of various grape seed oils

In this study, fatty acids, tocopherol contents, and phenolic compounds of the grape seed oils obtained with cold‐pressed and soxhlet extraction systems from several grape seeds were investigated. Linoleic acid contents of cold‐pressed oils varied between 59.61 and 72.13%. In addition, linoleic acid contents of oils obtained with soxhlet system changed between 58.51 and 71.17%. While α‐tocotrienol contents of oil samples extracted with cold‐pressed change between 13.21 and 34.71 mg/100 g, α‐tocotrienol contents of oils obtained by soxhlet system in all grape varieties varied between 8.89 and 33.89 mg/100 g. In addition, while ?‐tocotrienol contents of cold‐pressed oils range from 15.47 to 39.95 mg/100 g, ?‐tocotrienol contents of soxhlet extraction oils in all grape varieties were determined between 13.47 and 35.75 mg/100 g. Both systems' oils are rich in catechin, gallic acid, and chlorogenic acid. While catechin contents of cold‐pressed oils change between 427.80 and 847.14 mg/kg, catechin contents of soxhlet extraction oils in all grape seeds ranged from 537.11 to 836.88 mg/kg.

Practical applications

Cold‐pressed grape seed oil stands out as a suitable alternative to other commonly used vegetable oils because of its higher amounts of essential fatty acid, and many others bioactive compounds. Grape seed oil is being used in various fields such as cosmetics and cooking. Because the cold‐pressing does not need both heat and chemical treatments, cold‐pressed oils are being preferred for natural and safe food products by consumer. The consumption of vegetable oils rather than solid fats is crucial to maintaining health. As known, grape seed oil is an environment friendly oil due to a by‐product of wine and grape juice‐making processes.     

4 个葡萄品种葡萄籽冷榨油的性质与体外抗氧化活性

目的：评价4 种葡萄籽冷榨油脂肪酸组成、总酚含量及体外抗氧化活性。方法：收集单品种葡萄籽榨油,气相色谱法测定脂肪酸组成,Folin-酚法测定总酚含量,用1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picryhydrazyl,DPPH）法、2,2’-联氮双-（3-乙基苯并噻唑林-6-磺酸）二胺盐（2,2’-azinobis(3-ethylbenzothi azoline-6-sulfonic acid)ammonium salt,ABTS）法、Fenton反应和总抗氧化能力法分析其体外抗氧化能力。结果：4 个品种葡萄籽冷榨油均富含亚油酸（72.77%～77.36%）,不饱和脂肪酸含量为88.12%～91.06%;总酚含量为60.77～100.53 μg GAE/g,爱格丽葡萄籽冷榨油总酚含量最高,其次是北冰红、媚丽、赤霞珠葡萄籽冷榨油;4 个品种的葡萄籽冷榨油都有良好的清除DPPH自由基和ABTS＋·的能力,且随着葡萄籽冷榨油浓度的增大,清除能力增强,清除DPPH自由基的IC50变化范围为10.19～13.82 mg/mL,清除ABTS＋·的IC50变化范围为11.62～19.25 mg/mL;清除羟自由基（·OH）的能力范围为13.18～27.08 U/mL;总抗氧化能力范围为1.08～3.68 U/mL。4 个品种的葡萄籽冷榨油的体外抗氧化能力强弱顺序为爱格丽＞北冰红＞媚丽＞赤霞珠。     

Effects of exogenous abscisic acid on antioxidant capacities,anthocyanins, and flavonol contents of muscadine grape (Vitis rotundifolia) skins

The objective of this study was to evaluate the effect of exogenous abscisic acid (ABA) on the antioxidant capacity and phenolic content of muscadine grape skins (cvs. Noble and Alachua). ABA was applied on grapes during and after veraison. Average berry weight, total soluble solids (⁰Brix) and pH of the juice from both cultivars were not affected by the ABA treatment. Antioxidant capacity was enhanced by 38% and 18% in treated Noble at the first and second sampling, respectively. High performance liquid chromatography analysis revealed a significant increase in individual anthocyanins in treated Noble grapes at both sampling times. However, increase in the content of ellagic acid, myricetin, quercetin and kaempferol was observed at first sampling only. No effects of ABA treatment were seen in Alachua grapes. Our results indicate that exogenous application of ABA enhances the antioxidant capacity, anthocyanins and phenolic content of muscadine grapes but these effects may vary depending upon the cultivar.     

Total lipids content and fatty acid composition of seed oils from six pomegranate cultivars

The fatty acid composition of the seed oils of six pomegranate (Punica granatum L) cultivars was qualitatively and quantitatively determined by gas chromatography. The seeds contained oil in the range 51-152 mg kg^?1 dry matter. Intervarietal differences in fatty acid composition were demonstrated (fatty acid esters as % (w/w) total fatty acid esters. Sour varieties had the highest while sour-sweet varieties had the lowest oil content. Eleven fatty acids were identified. In all varieties, the predominant fatty acids were linoleic (25.2-38.6%) and oleic acid (24.8-35.5%) followed by palmitic (18.2-22.6%), stearic (6.9-10.4%) and linolenic acid (0.6-9.9%). To a lesser extent arachidic (1.1-3.4%) and palmitoleic acid (0.2-2.7%) were also found in all varieties. Lauric, myristic, behenic and lignoceric acids were rarely detected. As far as we know linolenic (18:3), arachidic (20:0), behenic (22:0) or lignoceric (24:0) acids have not been previously reported in the seed oils of edible pomegranate varieties. Lignoceric acid has not previously been found in pomegranate seed oil.     

Fatty Acid Composition of ‘Cerrado’ Seed Oils

The seeds of 28 species from ‘cerrado’, a typical savanna ecosystem of Brazil, were analysed for total lipid contents and fatty acid distribution. The seeds of 10 species presented contents above 150 g kg⁻¹, the highest yield reaching 335 g kg⁻¹. Distribution of fatty acids based on polyunsaturated compounds seems to be rare in seed oils from ‘cerrado’: only three seed oils were found to be based on linoleic acid and none on linolenic acid. Eight seed oils, four of them Fabales, presented palmitic acid as a dominant constituent. Half of the species presented oleic acid based seed oils. Two species stand out for unusual fatty acid distribution: Qualea grandiflora (Vochysiaceae) with 171 g kg⁻¹ of seed oil presenting 723 g kg⁻¹ of lauric acid and Serjania erecta (Sapindaceae) with 256 g kg⁻¹ of seed oil presenting 623 g kg⁻¹ of eicosenoic acid.     

Effect of essential oils treatment on the frozen storage stability of chub mackerel fillets

The effect of bay leaf (BLO), thyme (TO), rosemary (RO), black seed (BSO), sage (SO), grape seed (GSO), flaxseed (FSO) and lemon (LO) essential oil from vegetable extracted on lipid oxidation and some other quality parameter of frozen chub mackerel during frozen storage at ?20°C were examined over a period of 11 months. Taste, odour, texture and overall acceptability of control samples were given ‘unacceptable’ scores by the sixth month. Based primarily on sensory data, the shelf-lives of frozen chub mackerel were found 6 month for samples treated with oil of TO, RO, BSO, SO and LO and 7 month for samples treated with BLO, GSO and FSO. During the 11-months storing process of chub mackerel, the values of pH, total volatile basic nitrogen (TVB-N), trimethylamine nitrogen (TMA-N) both in control group samples and samples treated with oils did not reach to deterioration levels. Thiobarbitüric acid (TBA) and free fatty acid (FFA) values for all treatments remained lower than TBA and FFA values of control samples throughout the 11 month storage period. Particularly, thyme oil treatment is effective in delaying lipid oxidation. Bay leaf, rosemary, sage, lemon, flaxseed and grape seed oils were fallowed.