HPLC-DAD-ESIMS analysis of phenolic compounds in nectarines, peaches, and plums

The phenolic compounds of 25 peach, nectarine, and plum cultivars were studied and quantified by HPLC-DAD-ESIMS. Hydroxycinnamates, procyanidins, flavonols, and anthocyanins were detected and quantified. White and yellow flesh nectarines and peaches, and yellow and red plums, were analyzed at two different maturity stages with consideration of both peel and flesh tissues. HPLC-MS analyses allowed the identification of procyanidin dimers of the B- and A-types, as well as the presence of procyanidin trimers in plums. As a general rule, the peel tissues contained higher amounts of phenolics, and anthocyanins and flavonols were almost exclusively located in this tissue. No clear differences in the phenolic content of nectarines and peaches were detected or between white flesh and yellow flesh cultivars. There was no clear trend in phenolic content with ripening of the different cultivars. Some cultivars, however, had a very high phenolic content. For example, the white flesh nectarine cultivar Brite Pearl (350-460 mg/kg hydroxycinnamates and 430-550 mg/kg procyanidins in flesh) and the yellow flesh cv. Red Jim (180-190 mg/kg hydroxycinnamates and 210-330 mg/kg procyanidins in flesh), contained 10 times more phenolics than cultivars such as Fire Pearl (38-50 mg/kg hydroxycinnamates and 23-30 mg/kg procyanidins in flesh). Among white flesh peaches, cultivars Snow King (300-320 mg/kg hydroxycinnamates and 660-695 mg/kg procyanidins in flesh) and Snow Giant (125-130 mg/kg hydroxycinnamates and 520-540 mg/kg procyanidins in flesh) showed the highest content. The plum cultivars Black Beaut and Angeleno were especially rich in phenolics.     

Composition of phenolic compounds in a Portuguese pear (Pyrus communis L. var. S. Bartolomeu) and changes after sun-drying

The composition of phenolic compounds of a Portuguese pear cultivar (Pyrus communis L. var. S. Bartolomeu) was determined by HPLC after thioacidolysis. The average concentration of phenolic compounds in pear harvested at commercial maturity stage was 3.7 g per kg of fresh pulp. Procyanidins were the predominant phenolics (96%), with a mean degree of polymerization (mDP) of 13-44; hydroxycinnamic acids (2%), arbutin (0.8%), and catechins (0.7%) were also present. The most abundant monomer in the procyanidin structures was (-)-epicatechin (99%), which was found as extension and terminal units; (+)-catechin (1%) was found only as a terminal unit. Sun-drying of these pears caused a decrease of 64% (on a dry pulp basis) in the total amount of native phenolic compounds. Hydroxycinnamic acids and procyanidins showed the largest decrease; the B2 procyanidin was not found at all in the sun-dried pear. Less affected were arbutin and catechins. In the sun-dried pear, the procyanidins with high mDP became unextractable in the solvents used.     

From plums to prunes: influence of drying parameters on polyphenols and antioxidant activity

Prunes, which are industrially obtained by dehydrating fresh plums at 85-90 degrees C for 18 h, contain higher levels of phenolic compounds than most other fruits. Prune phenolics have shown beneficial effects on human health. Reports are available in the literature on ascorbic acid, phenol composition, and antioxidant activity of fresh plums and prunes, but there is a lack of publications on the influence of drying parameters on the phenolic compounds and antioxidant activity. A study was carried out on two plum cultivars using two sets of air-drying temperatures: (i) air temperature at 85 degrees C until 50% of prune moisture level and then the temperature was lowered to 70 degrees C; (ii) air temperature at 60 degrees C. Whole fresh and dried fruits were assessed for phenolics (catechins, hydroxycinnamic acids, anthocyanins, and flavonols), ascorbic acid, and antioxidant activity (all parameters were calculated on a dry matter basis). Analysis of data shows that chlorogenic and neochlorogenic acid changes were affected by both process parameters and cultivar. Drying destroyed anthocyanins, and there was a significant decrease in flavonols. Ascorbic acid was drastically reduced in relation to process temperature. The most striking result was that drying at 85 degrees C doubled antioxidant activity in both cultivars, while contradictory results were found for 60 degrees C processed plums.     

Characterization and quantitation of phenolic compounds in new apricot (Prunus armeniaca L.) varieties

Thirty-seven apricot varieties, including four new releases (Rojo Pasión, Murciana, Selene, and Dorada) obtained from different crosses between apricot varieties and three traditional Spanish cultivars (Currot, Mauricio, and Búlida), were separated according to flesh color into four groups: white, yellow, light orange, and orange (mean hue angles in flesh were 88.1, 85.0, 77.6, and 72.4, respectively). Four phenolic compound groups, procyanidins, hydroxycinnamic acid derivatives, flavonols, and anthocyanins, were identified by HPLC-MS/MS and individually quantified using HPLC-DAD. Chlorogenic and neochlorogenic acids, procyanidins B1, B2, and B4, and some procyanidin trimers, quercetin 3-rutinoside, kaempferol 3-rhamnosyl-hexoside and quercetin 3-acetyl-hexoside, cyanidin 3-rutinoside, and 3-glucoside, were detected and quantified in the skin and flesh of the different cultivars. The total phenolics content, quantified as the addition of the individual compounds quantified by HPLC, ranged between 32.6 and 160.0 mg 100 g(-1) of edible tissue. No correlation between the flesh color and the phenolic content of the different cultivars was observed.     

Role of the waxy skin layer in moisture loss during dehydration of prunes

Permeability studies of water through the skin and flesh of d'Agen plums have been carried out by radiotracer and PGSE NMR techniques as a function of moisture content. The results have shown that the diffusion coefficient of water through the skin layer increases as the fruit is dried at 70 degrees C or above. By contrast, the water diffusion through the fruit becomes more hindered as moisture is lost and structural collapse of the cell layers takes place. Values for diffusion coefficients of water at 21 degrees C through the fruit of 1 x 10(-)(9) m(2) s(-)(1) for fresh plums and 2.0 x 10(-)(10) m(2) s(-)(1) for fruit dried to 50% moisture content (wet basis) were found. Structural studies using scanning electron microscopy techniques were also performed. These showed that drying induces marked changes to the waxy skin layer of the fruit as well as the fruit flesh. These results are discussed in terms of the important role that the plum skin has in modulating the moisture loss process during dehydration, particularly at early stages of drying.     

Contribution of individual polyphenolics to total antioxidant capacity of plums

To study the effect of polyphenolics on antioxidant capacities of plums, the amounts of total phenolics, total flavonoids and individual phenolic compounds, and vitamin C equivalent antioxidant capacity (VCEAC) of eleven plum cultivars was determined. There was a good linear relationship between the amount of total phenolics and total antioxidant capacity (r2 = 0.9887). The amount of total flavonoids and total antioxidant capacity also showed a good correlation (r2 = 0.9653). Although the summation of individual antioxidant capacity was lower than the total antioxidant capacity of plum samples, there was a positive correlation (r2 = 0.9299) of total antioxidant capacity of plum samples with the sum of the VCEACs calculated from individual phenolics. Chlorogenic acids and glycosides of cyanidin, peonidin, and quercetin were major phenolics among eleven plum cultivars. The antioxidant capacity of chlorogenic acids and anthocyanins showed higher correlation (r2) of 0.7751 and 0.6616 to total VCEAC, respectively, than that of quercetin glycosides (r2 = 0.0279). Chlorogenic acids were a major source of antioxidant activity in plums, and the consumption of one serving (100 g) of plums can provide antioxidants equivalent to 144.4-889.6 mg of vitamin C.     

Antioxidant capacities,phenolic compounds,carotenoids, and vitamin C contents of nectarine,peach, and plum cultivars from California

Genotypic variation in composition and antioxidant activity was evaluated using 25 cultivars, 5 each of white-flesh nectarines, yellow-flesh nectarines, white-flesh peaches, yellow-flesh peaches, and plums, at the ripe (ready-to-eat) stage. The ranges of total ascorbic acid (vitamin C) (in mg/100 g of fresh weight) were 5-14 (white-flesh nectarines), 6-8 (yellow-flesh nectarines), 6-9 (white-flesh peaches), 4-13 (yellow-flesh peaches), and 3-10 (plums). Total carotenoids concentrations (in microg/100 g of fresh weight) were 7-14 (white-flesh nectarines), 80-186 (yellow-flesh nectarines), 7-20 (white-flesh peaches), 71-210 (yellow-flesh peaches), and 70-260 (plums). Total phenolics (in mg/100 g of fresh weight) were 14-102 (white-flesh nectarines), 18-54 (yellow-flesh nectarines), 28-111 (white-flesh peaches), 21-61 (yellow-flesh peaches), and 42-109 (plums). The contributions of phenolic compounds to antioxidant activity were much greater than those of vitamin C and carotenoids. There was a strong correlation (0.93-0.96) between total phenolics and antioxidant activity of nectarines, peaches, and plums.     

Characterization and estimation of proanthocyanidins and other phenolics in coffee pulp (Coffea arabica) by thiolysis-high-performance liquid chromatography

Fresh and 3-day-old coffee pulp of the Arabica variety were analyzed for polyphenol composition followed by characterization by two different methods. The first method consisted in subjecting coffee pulp powder to direct thiolysis. For the second method, coffee pulp was subjected to successive solvent extractions, followed by thiolysis. Quantification of phenolic compounds was then achieved by high-performance liquid chromatography (HPLC) analysis of thiolysis products. Four major classes of polyphenols were identified: flavan-3-ols (monomers and procyanidins), hydroxycinnamic acids, flavonols, and anthocyanidins. Differences in concentration of procyanidins were observed between fresh and 3-day-old coffee pulp. Constitutive units were mainly epicatechin, representing more than 90% of the proanthocyanidin units, with average degrees of polymerization in the range of 3.8-9.1. Monomer to hexamer units of flavan-3-ols from fresh coffee pulp were separated by normal-phase HPLC. Molecular size of oligomeric proanthocyanidins was obtained by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Results obtained confirm the presence of oligomers of the flavan-3-ol (-)-epicatechin.     

Phenolic composition of the Brazilian seedless table grape varieties BRS Clara and BRS Morena

The detailed phenolic composition (anthocyanins, flavonols, hydroxycinnamic acid derivatives, stilbenes, and flavan-3-ols) in the skin and flesh of the new BRS Clara and BRS Morena seedless table grapes has been studied using HPLC-DAD-ESI-MS/MS. The two grapes, especially BRS Morena, contained high amounts of phenolic compounds, mainly located in their skins and qualitatively not different from those found in Vitis vinifera grapes. In addition, BRS Morena (a teinturier variety) showed qualitatively different phenolic compositions in its skin and flesh, mainly affecting the anthocyanin and flavonol profiles. Consistent with high phenolic contents, high antioxidant capacity values were registered for both grape varieties, especially for BRS Morena. Proanthocyanidins and hydroxycinnamoyl-tartaric acids were the major phenolic compounds found in BRS Clara and were also important in BRS Morena, although anthocyanins were the main phenolic compounds in the latter case. These results suggest that the entire grapes, including the skin, may potentially possess properties that are beneficial to human health. In this context, the BRS Morena grape can be considered as a high resveratrol producer.     

Radical scavenging activities of peels and pulps from cv. Golden Delicious apples as related to their phenolic composition

The relationship between phenolic composition and radical scavenging activity of apple peel and pulp was investigated in fruit produced according to both organic and integrated agricultural methods. Apple tissue extracts were subjected to high-performance liquid chromatography separation, which showed that as compared with pulps, peels are richer in almost all of the quantified phenolics. Flavonols, flavanols, procyanidins, dihydrochalcones, and hydroxycinnamates were the identified phenolic classes in peel tissue, and the most abundant compounds were epicatechin, procyanidin B2, and phloridzin. Pulps were poorer in phytochemicals. Their major phenolics were procyanidins and hydroxycinnamates. Flavonols in amounts <20 mg kg(-1) fresh weight (fw) were also found. In both peels and pulps, integrated production samples were richer in polyphenols. Among the 14 compounds identified, only phloridzin had a tendency to appear higher in organic peels. The total antioxidant capacities (TAC) of extracts were evaluated using the 1,1-diphenyl-2-picrylhydrazyl radical assay and were expressed as Trolox equivalents. Integrated peels gave the highest TAC (18.56 mM kg(-1) fw), followed by organic peels (TAC = 14.96), integrated pulps (TAC = 7.12), and organic pulps (TAC = 6.28). In peels, the top contributors to the antioxidant activity were found to be flavonols, flavanols, and procyanidins, which accounted for about 90% of the total calculated activity whereas in pulps, the TAC was primarily derived from flavanols (monomers and polymers) together with hydroxycinnamates. A good correlation between the sum of polyphenols and the radical scavenging activities was found. Among the single classes of compounds, procyanidins (in peels and pulps) and flavonols (in peels) were statistically correlated to the TAC.     

Phenolic profile of Asturian (Spain) natural cider

The polyphenolic composition of natural ciders from the Asturian community (Spain), during 2 consecutive years, was analyzed by RP-HPLC and the photodiode-array detection system, without previous extraction (direct injection). A total of 16 phenolic compounds (catechol, tyrosol, protocatechuic acid, hydrocaffeic acid, chlorogenic acid, hydrocoumaric acid, ferulic acid, (-)-epicatechin, (+)-catechin, procyanidins B2 and B5, phloretin-2'-xyloglucoside, phloridzin, hyperin, avicularin, and quercitrin) were identified and quantified. A fourth quercetin derivative, one dihydrochalcone-related compound, two unknown procyanidins, three hydroxycinnamic derivatives, and two unknown compounds were also found. Among the low-molecular-mass polyphenols analyzed, hydrocaffeic acid was the most abundant compound, representing more than 80% of the total polyphenolic acids. Procyanidins were the most important family among the flavonoid compounds. Discriminant analysis was allowed to correctly classify more than 93% of the ciders, according to the harvest year; the most discriminant variables were an unknown procyanidin and quercitrin.     

Nutrients and antioxidant molecules in yellow plums (Prunus domestica L.) from conventional and organic productions: a comparative study

Yellow plums (Prunus domestica L) conventionally and organically grown in the same farm were selected to study the influence of different agronomic practices on antioxidant vitamins (ascorbic acid, vitamin E, beta-carotene) and phenolics (total polyphenols, phenolic acids, flavonols) concentration. Conventional plums were grown on tilled soil. Three organic cultivations were performed: tilled soil, soil covered with trifolium, and soil covered with natural meadow. Differences in macronutrients were marginal, whereas antioxidant vitamins and phenolic compounds concentration markedly differed among cultivations. Ascorbic acid, alpha-, gamma-tocopherols, and beta-carotene were higher in organic plums grown on soil covered with natural meadow. The highest phenolic acids content was detected in plums grown on soil covered with trifolium. Total polyphenols content was higher in conventional plums. Quercetin was higher in conventional plums, but myrecitin and kaempferol were higher in organic plums. Under the same cultivar and climate conditions, the type of soil management turned out of primary importance in influencing the concentration of health-promoting compounds.     

Characterization and quantitation of low and high molecular weight phenolic compounds in apple seeds

The phenolic constituents of seeds of 12 different apple cultivars were fractionated by sequential extraction with aqueous acetone (30:70, v/v) and ethyl acetate after hexane extraction of the lipids. Low molecular weight phenolic compounds were individually quantitated by RP-HPLC-DAD. The contents of extractable and nonextractable procyanidins were determined by applying RP-HPLC following thiolysis and n-butanol/HCl hydrolysis, respectively. As expected, the results revealed marked differences of the ethyl acetate extracts, aqueous acetone extracts, and insoluble residues with regard to contents and mean degrees of polymerization of procyanidins. Total phenolic contents in the defatted apple seed residues ranged between 18.4 and 99.8 mg/g. Phloridzin was the most abundant phenolic compound, representing 79-92% of monomeric polyphenols. Yields of phenolic compounds significantly differed among the cultivars under study, with seeds of cider apples generally being richer in phloridzin and catechins than seeds of dessert apple cultivars. This is the first study presenting comprehensive data on the contents of phenolic compounds in apple seeds comprising extractable and nonextractable procyanidins. Furthermore, the present work points out a strategy for the sustainable and complete exploitation of apple seeds as valuable agro-industrial byproducts, in particular as a rich source of phloridzin and antioxidant flavanols.     

Phenolic composition of kiwifruit juice

Phenolic compounds in kiwifruit pulp were separated and characterized by reversed-phase HPLC, and the effect of juice processing on the phenolic composition was studied. Fractionation of phenolic compounds was achieved through selective elution from C-18 cartridges prior to preconcentration and subsequent separation by HPLC. Strongly acidic compounds were identified as derivatives of coumaric and caffeic acids, including chlorogenic acid, protocatechuic acid, and a derivative of 3,4-dihydroxybenzoic acid. The weakly acidic fraction contained epicatechin, catechin, and procyanidins (B3, B2, or B4 and oligomers). Flavonols were present as the glycosides of quercetin (glucoside, rhamnoside, and rutinoside) and kaempferol (rhamnoside and rutinoside). Phenolic compounds were present, at levels of <1-7 mg/L, in clarified juice. The concentration of phenolics was highest after high-temperature short-time treatment (HTST) of juice. Hydrolysis of hydroxycinnamic acids occurred after enzyme addition and HTST treatment. The flavonol glycoside composition is the best identifier of kiwifruit juice.     

Almond (Prunus dulcis (Mill.) D.A. Webb) skins as a potential source of bioactive polyphenols

An exhaustive study of the phenolic composition of almond ( Prunus dulcis (Mill.) D.A. Webb) skins was carried out in order to evaluate their potential application as a functional food ingredient. Using the HPLC-DAD/ESI-MS technique, a total of 33 compounds corresponding to flavanols, flavonols, dihydroflavonols and flavanones, and other nonflavonoid compounds were identified. Peaks corresponding to another 23 structure-related compounds were also detected. MALDI-TOF MS was applied to characterize almond skin proanthocyanidins, revealing the existence of a series of A- and B-type procyanidins and propelargonidins up to heptamers, and A- and B-type prodelphinidins up to hexamers. Flavanols and flavonol glycosides were the most abundant phenolic compounds in almond skins, representing up to 38-57% and 14-35% of the total quantified phenolics, respectively. Due to their antioxidant properties, measured as oxygen-radical absorbance capacity (ORAC) at 0.398-0.500 mmol Trolox/g, almond skins can be considered as a value-added byproduct for elaborating dietary antioxidant ingredients.     

Quantification of polyphenolics and their antioxidant capacity in fresh plums

Total phenolics, total flavonoids, and antioxidant capacity of 11 cultivars of fresh plums were determined using spectrophotometric methods. Identification and quantification of individual polyphenolics were performed using reversed-phase high-performance liquid chromatography equipped with a diode array detector. The total phenolic contents of various cultivars widely varied from 125.0 to 372.6 mg/100 g expressed as gallic acid equivalents. The level of total flavonoids in fresh plums ranged between 64.8 and 257.5 mg/100 g expressed as catechin equivalents. Antioxidant capacity, expressed as vitamin C equivalent antioxidant capacity (VCEAC), ranged from 204.9 to 567.0 mg/100 g with an average of 290.9 mg/100 g of fresh weight. Cv. Beltsville Elite B70197 showed the highest amounts of total phenolics and total flavonoids and the highest VCEAC. A positive relationship (correlation coefficient r (2)() = 0.977) was presented between total phenolics and VCEAC, suggesting polyphenolics would play an important role in free radical scavenging. The level of IC(50) value of superoxide radical anion scavenging activity of the plum cultivars ranged from 13.4 to 45.7 mg of VCEAC/100 g. Neochlorogenic acid was the predominant polyphenolic among fresh plums tested. Flavonols found in plum were commonly quercetin derivatives. Rutin was the most predominant flavonol in plums. Various anthocyanins containing cyanidin aglycon and peonidin aglycon were commonly found in all plums except for cv. Mirabellier and NY 101.     

Superoxide radical scavenging activity of the major polyphenols in fresh plums

The effect of polyphenols among different varieties of plums on superoxide radical scavenging activity (SRSA) was studied by an enzymatic method and their IC(50) values were determined. We found that the SRSA levels of the polyphenols were closely related to their chemical structures; cyanidin showed the lowest IC(50) among the polyphenols examined, and aglycones are more effective than their glycosides. BY 69-339 cultivar exhibited the lowest IC(50) among the eleven plum cultivars, which means the highest antioxidant activity in scavenging superoxide radicals, followed by French Damson, Cacaks Best, Beltsville Elite B70197, Empress, Castleton, Stanley, NY 6, NY 101, Mirabellier, and NY 9. IC(50) values showed a higher correlation with total flavonoids (r (2) = 0.8699) than total phenolics (r (2) = 0.8355), which indicated that flavonoids might contribute to the total SRSA more directly than other polyphenols. Anthocyanins in plums appeared to be the major contributors to the total SRSA, except for two yellow cultivars having no anthocyanins. Chlorogenic acid was the predominant phenolic acid, and it also exhibited SRSA significantly in the range of 1.0 to 94.9%. Quercetins were the major flavonols in plums. However, they showed relatively low contribution to the total SRSA.     

Fractionation of polymeric procyanidins from lowbush blueberry and quantification of procyanidins in selected foods with an optimized normal-phase HPLC-MS fluorescent detection method

The polymeric procyanidins were fractionated from lowbush blueberry on a Sephadex LH-20 column. The degree of polymerization (DP) for the polymers was determined by thiolysis to be in a range of 19.9 to 114.1. Normal-phase HPLC analysis indicated that the polymeric procyanidins did not contain oligomeric procyanidins with DP < 10. The polymers eluted as a single peak at the end of the chromatogram. The normal-phase HPLC gradient was modified to improve the separation of procyanidin monomers through decamers and to elute all the polymers beyond those as a distinct peak. Monomers through decamers were quantified individually. All the polymers (DP > 10) were quantified using a mixture of purified polymers as an external standard. Polymers were found to be the dominant procyanidins in brown sorghum bran, cranberry, and blueberry. Thiolysis of the polymer peaks indicated that epicatechin was present as extension units in these foods, however, the composition of terminal units varied considerably between catechin and epicatechin, or an A-type dimer linkage in the case of cranberry.     

Phenolic composition of the edible parts (flesh and skin) of Bordô grape (Vitis labrusca) using HPLC-DAD-ESI-MS/MS

The aim of this study was the detailed characterization of the phenolic composition and the determination of the antioxidant activity of the Bord? grape (Vitis labrusca) cultivated in South Brazil. The edible parts of Bord? grapes (flesh and skin) contained 1130 mg/kg of total phenolic compounds (as gallic acid), mainly located in the skins. Anthocyanin content in the skins was high, largely as 3,5-diglucosides (1359 mg/kg, as malvidin 3,5-diglucoside). Total flavonols accounted for 154 μmol/kg, mainly located in the skins and with myricetin 3-glucoside as the principal flavonol in both grape parts. Very low amounts of flavan-3-ol monomers and dimers and low amounts of polymeric proanthocyanidins, with a composition similar to that reported for V. vinifera grape varieties, were found in Bord? grape skins. Hydroxycinnamic acid derivatives mainly derived from caffeic acid and were found in the skins in high amounts, ten times higher than in the flesh (total amount: 483 μmol/kg). Finally, the Bord? grape cultivar can be considered a high resveratrol producer (10.91 mg/kg) and also exhibited a high value of total antioxidant capacity (37.6 ± 1.0 mmol/kg, as Trolox).     

Effect of sample preparation on the measurement of sugars, organic acids, and polyphenols in apple fruit by mid-infrared spectroscopy

The objectives of this study were (i) to test different conditions of freezing, thawing, and grinding during sample preparation and (ii) to evaluate the possibility of using mid-infrared spectroscopy for analyzing the composition of sugars, organic acids, and polyphenols in apples. Seven commercial apple cultivars were chosen for their large variability in composition (total polyphenols from 406 to 1033 mg kg(-1) fresh weight). The different conditions of sample preparation affected only the phenolic compounds and not sugars or organic acids. The regression models of the mid-infrared spectra showed a good ability to estimate sugar and organic acid contents (R(2) ≥ 0.96), except for citric acid. Good predictions were obtained for total phenolic, flavan-3-ols, and procyanidins (R(2) ≥ 0.94) provided oxidation was avoided during sample preparation. A rapid and simple procedure was then proposed for phenolic compounds using sodium fluoride during sample homogenization at ambient temperature and freeze-drying before spectra acquisition.