HPLC–MS identification and quantification of flavonol glycosides in 28 wild and cultivated berry species

Berries and red fruits are rich dietary sources of polyphenols with reported health benefits. More than 50 different flavonols (glycosides of quercetin, myricetin, kaempferol, isorhamnetin, syringetin and laricitrin) have been detected and quantified with HPLC–MSⁿ in fruits of blueberry, bilberry, cranberry, lingonberry, eastern shadbush, Japanese wineberry, black mulberry, chokeberry, red, black and white currants, jostaberry, red and white gooseberry, hardy kiwifruit, goji berry, rowan, dog rose, Chinese and midland hawthorn, wild and cultivated species of blackberry, raspberry, strawberry and elderberry. The phenolic constituents and contents varied considerably among the analyzed berry species. Elderberry contained the highest amount of total flavonols (450–568 mg kg⁻¹ FW), followed by berry species, containing more than 200 mg kg⁻¹ FW of total: chokeberry (267 mg kg⁻¹), eastern shadbush (261 mg kg⁻¹), wild grown blackberry (260 mg kg⁻¹), rowanberry (232 mg kg⁻¹), american cranberry (213 mg kg⁻¹) and blackcurrants (204 mg kg⁻¹). Strawberry (10.5 mg kg⁻¹) and white currants (4.5 mg kg⁻¹) contained the lowest amount of total flavonols. Quercetins represent the highest percentage (46–100%) among flavonols in most analyzed berries. In wild strawberry and gooseberry the prevailing flavonols belong to the group of isorhamnetins (50–62%) and kaempferols, which represent the major part of flavonols in currants (49–66%). Myricetin glycosides could only be detected in chokeberry, rowanberry and species from the Grossulariaceae, and Adoxaceae family and Vaccinium genus. Wild strawberry and blackberry contained from 3- to 5-fold higher total flavonols than the cultivated one.     

Berry fruit teas: Phenolic composition and cytotoxic activity

A detailed phenolic composition analysis of dry chokeberry, bilberry and black currant fruit teas prepared on the most common ways — decoction and infusion, was performed, together with evaluation of cytotoxic activity. The most abundant in phenolics were chokeberry teas, followed by bilberry and black currant teas, while the highest anthocyanin amount was determined in bilberry samples. LC/DAD/MS method was used for identification of 17 anthocyanins, 11 flavonoids and 4 hydroxycinnamic acid derivatives. Quantitative analysis of investigated teas was carried out by HPLC analysis, and major phenolic compounds in berry fruit teas were chlorogenic acids, quercetin glycosides and anthocyanin glycosides. Berry teas were investigated for cytotoxic activity against cervix carcinoma, melanoma, colon, and chronic myelogenous leukemia cell lines, and chokeberry decoction was the most effective against all cell lines. According to the results obtained in this study, pure chokeberry, bilberry and black currant teas could be valuable sources of flavonoids and anthocyanins.     

CHEMICAL AND ANTIOXIDANT PROPERTIES OF CULTIVATED AND WILD FRAGARIA AND RUBUS BERRIES

ABSTRACT

Content of individual sugars, organic acids, total phenolics (TPH), some important phenolic compounds (kaempferol, myricetin, quercetin, and ellagic acid) and total antioxidant capacity (TAC) in the fruits of strawberry, raspberry and blackberry were studied. A comparison was made between cultivars and wild relatives of each species (Fragaria vesca L., Rubus idaeus L. and Rubus fruticosus L.). The main sugars found were fructose and glucose both in the fruits of wild species and the studied cultivars. Citric acid was determined to be the major organic acid in most of tested berries with the exception of blackberry cultivars, where malic acid was dominant. The content of individual phenolic compounds varied among the wild species, as well as among the studied cultivars. Ellagic acid content was higher in F. vesca, R. idaeus and R. fruticosus (122.5 µg/g FW, 12.71 µg/g FW and 61.7 µg/g FW, respectively) than that obtained in analyzed cultivars. Overall, TPH expressed higher values in the wild strawberry and blackberry species in comparison to the studied cultivars, and consequently, the highest levels of TAC were recorded in F. vesca (5.78 mg asc/g FW), followed by R. fruticosus (4.95 mg asc/g FW).

PRACTICAL APPLICATIONS

In recent years, both wild and cultivated berries have become very attractive for consumers because of potentially beneficial phytochemicals contained in these fruits. Fruit nutritional quality can be described by standard quality parameters (sugars and organic acids), and the analysis of antioxidant capacity influenced by specific related compounds. The importance of flavonoids and other phenolics have been suggested to play a preventive role in the development of cancer and heart disease. A significant positive correlation observed in this study between total phenolics and total antioxidant capacity indicate the need for the use of wild species in the breeding programs of small fruits, especially strawberry and blackberry, in order to increase their nutritive value and the health benefits.     

Solid-phase extraction of berries’ anthocyanins and evaluation of their antioxidative properties

Solid-phase extraction (SPE) was used to obtain anthocyanin-rich extracts from five berry species: chokeberry, elderberry, black currant, blackberry and blueberry. During SPE more than 94.4% of the sugars and more than 88.5% of the acids present in the crude extracts were separated. The SPE resulted in 90–95.6% anthocyanins recovery. The antioxidative properties of the anthocyanin-rich extracts were tested by measuring their oxygen radical absorption capacity (ORAC), hydroxyl radical averting capacity (HORAC), total peroxyl radical trapping antioxidant parameter (TRAP), scavenging of nitric oxide and inhibition of lipid peroxidation. Elderberry extract revealed the highest ORAC value of 5783 μmol TE/g. Chokeberry extract was the most potent inhibitor of lipid peroxidation and had the highest TRAP value of 4051 μmol TE/g. Blueberry extract had the highest HORAC result – 1293 μmol GAE/g and was the most powerful scavenger of NO. The high antioxidant activity according to all antioxidant assays revealed opportunities to apply these preparations as antioxidants.     

Wild Prunus Fruit Species as a Rich Source of Bioactive Compounds

Sugars, organic acids, carotenoids, tocopherols, chlorophylls, and phenolic compounds were quantified in fruit of 4 wild growing Prunus species (wild cherry, bird cherry, blackthorn, and mahaleb cherry) using HPLC‐DAD‐MSn. In wild Prunus, the major sugars were glucose and fructose, whereas malic and citric acids dominated among organic acids. The most abundant classes of phenolic compounds in the analyzed fruit species were anthocyanins, flavonols, derivatives of cinnamic acids, and flavanols. Two major groups of anthocyanins measured in Prunus fruits were cyanidin‐3‐rutinoside and cyanidin‐3‐glucoside. Flavonols were represented by 19 derivatives of quercetin, 10 derivatives of kaempferol, and 2 derivatives of isorhamnetin. The highest total flavonol content was measured in mahaleb cherry and bird cherry, followed by blackthorn and wild cherry fruit. Total phenolic content varied from 2373 (wild cherry) to 11053 mg GAE per kg (bird cherry) and ferric reducing antioxidant power antioxidant activity from 7.26 to 31.54 mM trolox equivalents per kg fruits.     

5种野生浆果的体外抗氧化和抗细胞增殖活性

选取东北大兴安岭地区的特色浆果花楸(Sorbus pohuashanensis H.)、蓝莓(Vaccinium uliginosum L.)、蓝靛果(Lonicera caerulea L.)、沙棘(Hippophae rhamnoides L.)、草莓(Fragaria orientalis Los.)的成熟果实,采用80% 丙酮提取,分别测定多酚、黄酮、花色苷含量,并研究其抗氧化活性和抗细胞增殖活性。结果表明：花楸的多酚和黄酮含量最高;蓝靛果的花色苷含量最高,为(367.5 ± 8.7)g/100g;5 种浆果都具有较好的体外抗氧化活性,花楸显示了最高的抑制癌细胞增殖效应。     

Berry components inhibit α-glucosidase in vitro: Synergies between acarbose and polyphenols from black currant and rowanberry

Polyphenol-rich extracts from certain berries inhibited α-glucosidase activity in vitro. The two most effective berry extracts, from black currant and rowanberry, inhibited α-glucosidase with IC(50) values respectively of 20 and 30μg GAE/ml and were as effective as the pharmaceutical inhibitor, acarbose. These berry extracts differed greatly in their polyphenol composition: black currant was dominated by anthocyanins (～70% of total) whereas rowanberry was enriched in chlorogenic acids (65% total) and had low levels of anthocyanins. Both black currant and rowanberry extracts potentiated the inhibition caused by acarbose and could replace the inhibition lost by reducing the acarbose dose. However, no additive effects were noted when black currant and rowanberry extracts were added in combination. The mechanisms underlying the synergy between acarbose and the berry polyphenols and the lack of synergy between the berry components are discussed. These extracts exhibited the potential to replace acarbose (or reduce the dose required) in its current clinical use in improving post-prandial glycaemic control in type 2 diabetics. As a result, these polyphenols may offer a dietary means for type 2 diabetics to exercise glycaemic control.     

Developmental changes of carbohydrates,organic acids,amino acids,and phenolic compounds in ‘Honeycrisp’ apple flesh

The developmental changes of carbohydrates, organic acids, amino acids and phenolic compounds in ‘Honeycrisp’ apple flesh were investigated using GC–MS and HPLC. A total of 12 carbohydrates, 8 organic acids, 20 amino acids, and 18 phenolic compounds were identified and quantified. Each metabolite showed characteristic changes during fruit development, but in general, concentrations of most sugars and sugar alcohols either increased or remained unchanged whereas concentrations of most organic acids, amino acids and phenolic compounds decreased with fruit development, indicating that most sugars and sugar alcohols are synthesised and/or accumulate at a faster or similar rate relative to fruit growth whereas organic acids, amino acids and phenolics are synthesised and/or accumulate at a slower rate relative to fruit growth. On a whole fruit basis, the content of most metabolites increased with fruit development. In the flesh of mature ‘Honeycrisp’ apple, fructose and sucrose and sorbitol are the major sugars and sugar alcohol; malic acid is the major organic acid; aspartic acid, asparagine, glutamic acid, proline, threonine and γ-aminobutyric acid are the major amino acids; and procyanidin B1, procyanidin B2, chlorogenic acid, catechin and epicatechin are the major phenolic compounds, respectively.     

Complex approach to evaluation of natural quality of coloured juices

A novel multidimensional approach to estimation of fruit and berry colored juice authenticity is proposed. It includes determination of quality (determination of 5-hydroxymethylfurfurol) and authenticity (determination of organic acids, hydrocarbons, anthocyanin pigments and artificial colorants) parameters. Both experimental and literature data on chemical composition of 12 colored fruits and berries are summarized. New composition characteristics of pomegranate, bilberry, red currant, blackberry and lingonberry are presented.     

Berry antioxidants: small fruits providing large benefits

Small berry fruits are consumed because of their attractive colour and special taste, and are considered one of the richest sources of natural antioxidants. Their consumption has been linked to the prevention of some chronic and degenerative diseases. The term ‘berry fruits’ encompasses the so‐called ‘soft fruits’, primarily strawberry, currants, gooseberry, blackberry, raspberry, blueberry and cranberry. The objective of this review is to highlight the nutraceutical value of berries and to summarize the factors affecting berry fruit antioxidants. Particular attention is given to postharvest and processing operation factors that may affect fruit phytochemical content. The structure–antioxidant relationships for phenolic compounds – the main group of antioxidants in this fruit group – are presented and major areas for future research are identified. © 2013 Society of Chemical Industry     

Comparative study of primary and secondary metabolites in apricot (Prunus armeniaca L.) cultivars

BACKGROUND: Apricot is one of the most popular Prunus species grown in the temperate zone and great variation in the contents of primary and secondary metabolites has been reported in its fruit. However, little is known on the phenolic profiles and antioxidant potential of important local cultivars. The objective of the study was to identify valuable parental lines for breeding of cultivars with high health‐promoting effects. RESULTS: Primary metabolites (sugars, organic acids) and secondary metabolites (phenolics) were quantified by high‐performance liquid chromatography–mass spectrometry (HPLC/MS) in fruit of 13 apricot cultivars: total sugars ranged from 59.2 to 212.5 g kg^?1 fresh weight (FW) and total organic acids from 4.2 to 20.8 g kg^?1 FW. Four hydroxycinnamic acids and three flavonols were quantified; their content was significantly higher in skin compared to pulp. Similarly, antioxidative potential was significantly higher in skin and ranged from 125.4 to 726.5 mg ascorbic acid equivalents kg^?1 FW. A positive correlation between total phenolic content and antioxidant potential was determined. Multivariate analysis subdivided the cultivars into five major groups, mostly influenced by the content of individual and total phenolics as well as the antioxidant potential of apricot skin and pulp. CONCLUSION: In regard to high phenolic content in connection with antioxidant potential, three cultivars stand out as the richest in analyzed quality parameters: ‘Mula Sadik’, ‘Bergeron’ and ‘Chuan Zhi Hong’. They also contained high amounts of primary metabolites and should be promoted for further planting and included in apricot breeding programs. Copyright © 2010 Society of Chemical Industry     

<Emphasis Type="Italic">Aronia melanocarpa</Emphasis> phenolics and their antioxidant activity

The purpose of the present study was to evaluate small and high molecular phenolics (tannins) and antioxidant activity of Aronia melanocarpa berries, juice and pomace in order to find new potential sources of natural antioxidants. The fruits of Aronia melanocarpa Elliot were collected in the middle of October at a plantation near Wroclaw, Poland. The pomace has a much higher content of phenolics in comparision to juice and fruits. Results showed that polymeric proanthocyanins, predominantly of (−)epicatechin, are the major class of polyphenolic compounds in chokeberry, represent 66% of fruits polyphenols. The average concentration ranged from 1578.79 mg/100 g of DW for chokeberry juice up to 8191.58 mg/100 g in pomace. The concentration of phenolic acids (chlorogenic and neochlorogenic acids) in juice was higher than in pomace. Anthocyanins in Aronia melanocarpa are second phenolic compound group and represent about 25% of total polyphenols, mixture of four different cyanidin glycosides: 3-galactoside, 3-glucoside, 3-arabinoside and 3-xyloside. The higher antioxidant activity expressed as TEAC was measured in pomace >fruit >juice.     

Antioxidant activity and antimicrobial effect of berry phenolics--a Finnish perspective

In Finland, berries are part of the traditional diet significantly contributing to the intake of flavonoids and other phenolic compounds. Compositional data on phenolic compounds in berries has been rapidly accumulating and included in the national food composition database. Among the different bioactive substances in berries, phenolic compounds including flavonoids, tannins, and phenolic acids have received considerable interest due to their effects in food and health. A great amount of in vitro evidence exists showing that berry phenolics are powerful antioxidants. However, the antioxidant effect of berry phenolics is strongly dependent on the choice of berry raw material, as the antioxidant activity differs between the different phenolic constituents, including anthocyanins, ellagitannins, and proanthocyanidins. In foods, the antioxidant effect is also influenced by the structure of food. Tannin-containing berries exhibit antimicrobial properties against pathogenic bacteria, thus offering many new applications for food industry. Much of the interest in berry phenolics has focused on cranberries and both cultivated and wild blueberries, although also other berries including black currants, cloudberries, lingonberries, and red raspberries possess promising bioactivities that may have relevance to human health. Antioxidant activity of berry phenolics, in addition to other mechanisms, may contribute to human health, but the possible relationship remains yet to be scientifically substantiated.     

Effects of Food Processing on Blueberry Antiproliferation and Antioxidant Activity

Cultivated highbush (Vaccinium corymbosum L.) and wild lowbush (Vaccinium angustifolium Ait.) blueberries are excellent sources of phytochemicals that are believed to have significant biological activity. The objective of this study was to determine whether incorporation of blueberries into food products affects their phenolic content or antioxidant and antiproliferation activity. Several blueberry fruit‐containing products including fresh, individually quick frozen (IQF), freeze‐dried, spray‐dried, heat‐dried, cooked, juice concentrate, pie filling, and jam were fractionated to remove sugars and isolate groups of phytochemicals based on solubility. The fractions were analyzed for total phenolics and assayed for ferric reducing antioxidant power (FRAP) antioxidant activity, 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical scavenging activity, and hepa‐1c1c7 antiproliferative activities. For both cultivated and wild berries, fresh and IQF berries had the highest total phenols, antioxidant activity, and antiproliferation activity. Whole freeze‐dried wild blueberries also retained significant antiproliferative activity in 2 fractions eluted with acetone (fraction 4, 4% of control cell growth at 20 (μg/mL) and 50% aqueous acetone (fraction 5, 69% of control cell growth at 20 (μg/mL) and ranked close to the activities recorded for fresh (30% of control cell growth at 20 (μg/mL for fraction 5) and IQF whole fruit (27% of control cell growth at 20 (μg/mL for fraction 5). Products that were heat‐processed retained most of the antioxidant activity and total phenolics found in unprocessed whole fruit. However, the heat‐treated products lacked or had diminished antiproliferation activity, suggesting that although products may be high in phenolic compounds and antioxidant activity, some forms of bioactivity may be compromised by harsh processing methods.     

五种寒地果树果实的多酚含量、抗氧化活性及抗α-淀粉酶活性分析

为了探究寒地果树资源果实中的营养价值及体外活性,本试验测定了软枣猕猴桃、黑果腺肋花楸、草原樱桃、白穗醋栗和蔓越莓五种果实的总酚、总花色苷含量,比较了其抗氧化及抗α-淀粉酶活性,并进行了相关性分析,结果表明:不同寒地果实的总酚、总花色苷含量之间存在显著差异(P<0.05)。总酚含量为13.21~66.43 mg GAE/100 g,其中黑果腺肋花楸含量最高,白穗醋栗次之;总花色苷含量为0.15~46.45 mg C3G/100 g,黑果腺肋花楸含量最高,草原樱桃次之。另外5种果实的抗氧化能力也有所差异。用DPPH法、ABTS法、ORAC法、FRAP法测得白穗醋栗的抗氧化活性最强。用高通量淀粉浊度法测得白穗醋栗的抗α-淀粉酶活性最强,IC₅₀为0.122 mg/mL,高于阿卡波糖(0.16 mg/mL),其AE值为2034.18 mmol AE/g,高于阿卡波糖(1550 mmoL AE/g),且是其他果实的42~127倍。相关性分析发现五种寒地果实的总酚、总花色苷含量和抗氧化活性之间存在极显著正相关(P<0.01),而与抗α-淀粉酶活性之间无显著性相关关系。     

Identification and quantitative analysis of sugars and non-volatile organic acids in Chinese gooseberry fruit (Actinidia chinensis Planch.).

The free sugars and non-volatile organic acids in Chinese gooseberry fruit were extracted and separated by lead salt and ion-exchange procedures. Sugars and acids were identified and quantitatively determined by gas-liquid chromatography (g.l.c.) of their trimethylsilyl (TMS) derivatives. Glucose, fructose, sucrose and trace amounts of sorbitol accounted for the total sugars. Citric, quinic and malic are the major acids, the outstanding feature being the high content of quinic acid in mature fruit. Small amounts of phosphoric, ascorbic, glucuronic, galacturonic, oxalic, succinic, fumaric, oxalacetic and p-coumaric acids are also present. Qualitative and quantitative analysis of acids prepared as lead salts agreed closely with the determination of acids isolated by the ion-exchange procedure. Identifications of sugars and acids were confirmed by paper and thin-layer chromatography. Reducing sugars, total sugars and titratable acidity were determined by AOAC methods and the results compared with those from g.l.c.     

中国野生葡萄果实基本品质、酚类物质含量及其抗氧化活性分析

中国野生葡萄资源丰富,但对其利用有限;为揭示不同种野生葡萄的品质差异及进一步利用奠定理论基础,本研究对中国野生葡萄的10个种(46个株系)和不同对照品种(4份材料)的基本品质、酚类物质含量及其抗氧化活性指标进行评价。结果表明:1)除刺葡萄外,中国野生葡萄的单果质量小于‘赤霞珠’;中国野生葡萄的皮果比高于‘赤霞珠’。2)中国野生葡萄和对照之间糖的组成相似,果糖与葡萄糖所占的比例没有差异,总糖含量显著低于‘赤霞珠’和‘户太8号’;除刺葡萄外,中国野生葡萄的总酸含量显著高于对照‘赤霞珠’和‘户太8号’,中国野生葡萄的酒石酸/苹果酸的比值低于‘赤霞珠’,说明中国野生葡萄属于高苹果酸型;河岸葡萄和圆叶葡萄的糖酸含量与中国野生葡萄的糖酸含量较接近;‘赤霞珠’和‘户太8号’属于高糖低酸型,中国野生葡萄属于低糖高酸型。3)中国野生葡萄的抗氧化活性和酚类物质的含量变异较大,大部分的野生葡萄与‘赤霞珠’的酚类物质含量和抗氧化活相近,但中国野生葡萄的花色苷含量高于对照的花色苷含量。综上所述,中国野生葡萄中的燕山葡萄、蘡薁葡萄和刺葡萄的品质较好,其与‘赤霞珠’和‘户太8号’的糖酸组分、酚类物质含量及抗氧化活性接近。     

Effect of methyl jasmonate on quality and antioxidant activity of postharvest loquat fruit

BACKGOUND: Loquat fruit is rich in natural antioxidants and has shown a remarkably high antioxidant activity. To search for an effective method for maintaining or even improving antioxidant activity during postharvest storage, we investigated the effect of 10 µmol L^?1 methyl jasmonate (MeJA) treatment on levels of major individual sugars and organic acids, total phenolics, total carotenoids, total flavonoids and antioxidant activity in loquat fruit during storage at 1 °C for 35 days. RESULTS: The MeJA‐treated fruit exhibited significantly lower levels of respiration rate, ethylene production, phenylalanine ammonia‐lyase and polyphenol oxidase activities, and higher levels of sugars, organic acids, total phenolics and total flavonoids than control fruit. Meanwhile, the treatment also maintained significantly higher antioxidant activity as measured by the scavenging capacity against 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), superoxide and hydroxyl radicals, and by the reducing power test compared to the control. There was a significant positive linear relationship between total phenolic content and DPPH radical scavenging activity. CONCLUSION: MeJA treatment can improve the quality and functional properties of harvested loquat fruit by maintaining a higher level of antioxidants and enhancing antioxidant activity. Copyright © 2009 Society of Chemical Industry     

Screening of selected flavonoids and phenolic acids in 19 berries

Selected flavonoids (kaempferol, quercetin, myricetin) and phenolic acids (p-coumaric, caffeic, ferulic, p-hydroxybenzoic, gallic and ellagic acids) were simultaneously detected from 19 berries using a simple High Performance Liquid Chromatographic (HPLC) method. These phenolics have been proposed to have beneficial effects on health as antioxidants and anticarcinogens. Marked differences were observed in the phenolic profiles among the berries, with certain similarities within families and genera. The major phenolic compound analyzed in the genus Vaccinium was quercetin in lingonberry and cranberry, and its level was high also in blueberries and bilberry. In the genus Ribes, quercetin was the main compound in gooseberry, red currant and black currant. Ellagic acid was the main phenolic compound in the berries of the genus Rubus (red raspberry, Arctic bramble and cloudberry) and genus Fragaria (strawberry). Our data suggest that berries have potential as good dietary sources of quercetin or ellagic acid.     

Phytochemicals in fruits of two Prunus domestica L. plum cultivars during ripening

BACKGROUND: Plums are a rich source of nutritive and bioactive compounds. The objective was to evaluate the impact of fruit tissue (flesh and peel), ripening stage and cultivar on the composition of phytochemicals in plums (Prunus domestica L.) and to analyse the relation between total antioxidant activity (TAA) and the measured variables. RESULTS: The content of total sugars and total organic acids was higher in flesh but total phenolic content (TPC) and TAA was statistically higher in plum peel. The composition of sugars depended on fruit tissue, ripening stage and the cultivar. Ripening and fruit tissue affected the composition and concentrations of organic acids. TPC of ‘Haganta’ peel increased during ripening but there was no significant time trend for ‘Jojo’ peel and flesh of both cultivars. The composition of phenolics in peel depended on the cultivar, but in flesh on the cultivar and ripening. Ripening resulted in increased total anthocyanins and TAA in peel. TAA_peel was positively correlated with TPC_peel, total anthocyanins_peel, cyanidin 3‐glucoside_peel and peonidin 3‐rutinoside_peel in the cultivar ‘Haganta’. CONCLUSION: The composition of phytochemicals in P. domestica fruit depends on fruit tissue, ripening stage and the cultivar. Peel contributes to the TAA on average 20 times more than flesh. © 2012 Society of Chemical Industry