HPLC-DAD-ESIMS analysis of phenolic compounds in bayberries (Myrica rubra Sieb. et Zucc.)

Qualitative analysis of the ethyl acetate extracts from three bayberry cultivars, Xiangshan, Biqi and Dongkui, was performed by means of a hyphenated technique of HPLC coupled to photodiode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESIMS). Three phenolic compounds were identified (gallic acid, protocatechuic acid, and quercetin 3-glucoside) and seven others (two myricetin hexoside and two myricetin deoxyhexoside derivatives; quercetin hexoside and quercetin deoxyhexoside derivatives; kaempferol hexoside derivative) partially identified. Quantification of phenolic compounds was performed by HPLC-DAD, which revealed that gallic acid (2.6–7.0 mg/kg FW) was the major phenolic acid in all analysed cultivars. Myricetin glycosides (71.1 mg/kg FW) were the major flavonol glycosides in cultivar Xiangshan and quercetin glycosides (117.8 mg/kg FW) were the major ones in cultivar Biqi. Cultivar Dongkui had medium contents of quercetin glycosides (48.0 mg/kg FW) and myricetin glycosides (53.2 mg/kg FW). Kaempferol glyosides (3.1–4.6 mg/kg FW) were the lowest contents of flavonol glycosides in the assayed bayberries. These results are relevant not only from a nutritional point of view, but also in the control of color stability and haze formation during bayberry juice processing and storage.     

Identification and characterisation of low-molecular-weight phenolic compounds in bayberry (Myrica rubra Sieb. et Zucc.) leaves by HPLC-DAD and HPLC-UV-ESIMS

Leaves of each of two bayberry cultivars, Biqi and Dongkui, were divided into three categories by age, namely immature, intermediate, and mature. Phenolic compounds were analysed by the methods of HPLC-DAD and HPLC-UV-ESIMS. Gallic acid and EGCG were identified positively, and 13 other compounds (flavan-3-ol monomers, prodelphinidin oligomers, and flavonol glycosides) were partially identified. Gallic acid (7.5–87.8 mg/100 g) was the only phenolic acid detected and flavan-3-ols were abundant. Myricetin deoxyhexoside (535.4–853.0 mg/100 g) was the major flavonol glycoside. Among the three categories, immature leaves of both cultivars recorded the highest level of total phenolics, irrespective of whether they were measured by the Folin–Ciocalteu method (19404.0 mg/100 g in Biqi and 19626.0 mg/100 g in Dongkui) or as the sum of individual phenolic compounds (2255.9 mg/100 g in Biqi and 1797.1 mg/100 g in Dongkui). The results showed that bayberry leaves are a potentially rich source of beneficial phenolics.     

Chemical composition,anthocyanins, non-anthocyanin phenolics and antioxidant activity of wild bilberry (Vaccinium meridionale Swartz) from Colombia

Berries of Vaccinium meridionale Swartz native to Colombia were analysed for chemical composition, total phenolic content, anthocyanin content, and antioxidant activity. In addition, high-performance liquid chromatography with photodiode array detection (HPLC–DAD) and HPLC-electrospray ionisation tandem mass spectrometry (ESI–MS/MS) were used to determine anthocyanin and phenolic composition. Anthocyanin content was 329.0 ± 28.0 mg cyanidin 3-glucoside equivalents/100 g (fresh weight) FW and total phenolic content was 758.6 ± 62.3 mg gallic acid equivalent/100 g FW. Cyanidin 3-galactoside was the major anthocyanin while the most abundant non-anthocyanin phenolic was chlorogenic acid.     

Fruit quality and bioactive compounds relevant to human health of sweet cherry (Prunus avium L.) cultivars grown in Italy

The fruit quality characteristics, phenolic compounds and antioxidant capacities of 24 sweet cherry (Prunus avium L.) cultivars grown on the mountainsides of the Etna volcano (Sicily, Italy) were evaluated. High-performance liquid chromatographic methods were used to identify and quantify sugars, organic acids and phenolics. A total of seven phenolic compounds were characterised as hydroxycinnamic acid derivatives (neochlorogenic acid, p-coumaroylquinic acid and chlorogenic acid) and anthocyanins (cyanidin 3-glucoside, cyanidin 3-rutinoside, pelargonidin 3-rutinoside and peonidin 3-rutinoside). The total anthocyanin content ranged from 6.21 to 94.20 mg cyanidin 3-glucoside equivalents/100 g fresh weight (FW), while the total phenol content ranged from 84.96 to 162.21 mg gallic acid equivalents/100 g FW. The oxygen radical absorbance capacity (ORAC) assay indicated that fruit of all genotypes possessed considerable antioxidant activity. The high level of phenolic compounds and antioxidant capacity of some sweet cherry fruits implied that they might be sources of bioactive compounds that are relevant to human health.     

Preliminary characterisation of cornelian cherry (Cornusmas L.) genotypes for their physico-chemical properties

Fruit weight, antioxidant capacity, total anthocyanins, total phenolics, ascorbic acid, soluble solid content (SSC), reducing sugar and acidity of a number of selected cornelian cherry (Cornusmas L.) genotypes of varied pigmentation were investigated. Two methods, namely β-carotene bleaching and ferric reducing antioxidant power (FRAP) were used to determine total antioxidant capacity, while Folin–Ciocalteu reagent was used to determine total phenols. Fruit weight, SSC and ascorbic acid content of genotypes were 2.09–9.17; 12.53–21.17% and 29–112 mg/100 g, respectively. Antioxidant activity and total phenolic content varied among genotypes and 44-18 genotype had the highest antioxidant capacity using both methods. This genotype also had the highest total phenolic (74.8 mg GAE/g DW) and total anthocyanin (115 mg cyanidin-3-glucoside equivalents /100 g FW) content. There are linear relationships between antioxidant capacities and total phenols. The present study demonstrates the potential of certain cornelian cherry genotypes, notably 44-18, for improvement of nutritional value through germplasm enhancement programmes.     

Phenolic and antioxidant composition of by-products from the cider industry: Apple pomace

Eleven different cider apple pomaces (six single-cultivar and five from the cider-making industry) have been analysed for low molecular phenolic profiles and antioxidant capacity. The Folin index ranged between 2.3 and 15.1 g gallic acid per kg of dry matter. Major phenols were flavanols, dihydrochalcones (phloridzin and phloretin-2′-xyloglucoside), flavonols and cinnamic acids (chlorogenic and caffeic acids). The group of single-cultivar pomaces had higher contents of chlorogenic acid, (-)-epicatechin, procyanidin B2 and dihydrochalcones, whereas the industrial samples presented higher amounts of up to four unknown compounds, with absorption maxima between 256 and 284 nm. The antioxidant capacity of apple pomace, as determined by the DPPH and FRAP assays, was between 4.4 and 16.0 g ascorbic acid per kg of dry matter, thus confirming that apple pomace is a valuable source of antioxidants. PLSR analysis gave reliable mathematical models which allowed to predict the antioxidant activity of apple pomace as a function of the phenolic profile. The variables with the higher modelling power were phloridzin > procyanidin B2 > rutin + isoquercitrin > protocatechuic acid > hyperin.     

Antioxidant activity and phenolics of an endophytic Xylaria sp. from Ginkgo biloba

The objective of this study was to evaluate the antioxidant activity of cultivated fruiting bodies of an endophytic Xylaria sp. (strain number YX-28), from Ginkgo biloba. The results indicated that the methanol extract exhibited strong antioxidant capacity in both 2,2-diphenyl-1-picrylhydrazyl (DPPH) analysis and β-carotene–linoleic acid model system. Total phenolic and flavonoid contents extracted by different solvents were determined using Folin–Ciocalteu procedure and the flavonoid–aluminium method. The results showed that total phenolic and flavonoid contents were the highest in methanol extract (54.51 ± 1.05 mg gallic acid equivalent/g dry weight and 86.76 ± 0.58 mg rutin equivalent/g dry weight), while the hexane extract was the lowest (9.71 ± 0.57 mg GAE/g dw and 10.14 ± 0.76 mg RE/g dw, respectively). The correlation coefficients from regression analysis showed a positive relationship between total phenolic content in the extracts and DPPH activity (R² = 0.7336), as well as between total flavonoid content and DPPH activity (R² = 0.9392). Furthermore, GC/MS method was used to confirm the presence of phenolics with antioxidant activity in the methanol extract and resulted in the identification of 41 compounds, esters, phenolics, alkanes, carboxylates and alcohols being the main components. In conclusion, cultivated fruiting bodies of Xylaria sp.YX-28 may have potential as natural antioxidant.     

Physicochemical and antioxidative properties of red and black rice varieties from Thailand,China and Sri Lanka

Nine red and three black rice varieties from Thailand, China and Sri Lanka were analysed to determine their proximate composition and their physicochemical and antioxidant properties. Four groups of rice varieties with different amylose contents were identified. Cyanidin 3-glucoside and peonoidin 3-glucoside were confirmed as the dominant anthocyanins in black rice varieties with contents ranging from 19.4 to 140.8 mg/100 g DM and 11.1–12.8 mg/100 g DM, respectively. Total phenolic content (TPC) differed significantly between the varieties, but not between the colours. Highest TPC was found in the red Thai rice Bahng Gawk (BG) with 691 FA equivalent mg/100 g DM, which showed as well the highest antioxidant properties. In red varieties, the major phenolic acids in the free form were ferulic, protocatechuic and vanillic acid, whereas in black varieties protocatechuic acid was dominant followed by vanillic and ferulic acid. In the bound form, ferulic acid was predominant in both colours, where contents differed significantly, followed by p-coumaric and vanillic acid. The antioxidative capacity did not differ significantly between both colours but amongst genotypes. Antioxidant capacity of rice varieties ranged within 0.9–8.1 mmol Fe(II)/100 g DM for FRAP and 2.1–12.3 mmol TEAC/100 g DM. DPPH scavenging ability ranged from 13.0% to 76.4% remaining DPPH.     

Inhibitory effect of raspberries on starch digestive enzyme and their antioxidant properties and phenolic composition

Seven primocane fall-bearing raspberry (Rubus idaeus L.) cultivars, Nova (red), Dinkum (red), Heritage (red), Autumn Britten (red), Josephine, Anne (yellow), Fall Gold (yellow) were analysed for potential health promoting properties including their inhibitory effect on starch and fat digestive enzymes, antioxidant activities, and phenolic composition. The tested raspberry extracts showed no detectable inhibition of pancreatic α-amylase and lipase. However, all the extracts exhibited potent inhibition of α-glucosidase with IC₅₀ from 16.8 to 34.2 μg/mL. Four phenolic compounds, ellagic acid, cyanidin-diglucoside, pelargonidin-3-rutinoside, and catechin were identified as the active α-glucosidase inhibitors. The raspberry extracts also possessed significant antioxidant activities with oxygen radical absorbance capacities (ORAC) ranging from 136.7 to 205.2 μmol Trolox equivalents (TE)/g dry weight fruit and DPPH radical scavenging activities from 305 to 351 μmol TE/g. The total phenolic content of raspberry cultivars varied significantly from 40.9 to 98.5 mg of gallic acid equivalents/g dry weight. The anthocyanin content varied widely from 0.1 to 9.5 mg cyanidin 3-glucoside equivalents/g. Nine phenolic acids were quantified in raspberries and their total amounts varied from 157.3 to 713.5 μg/g. The enzyme inhibition and antioxidant properties of raspberry cultivars were not correlated with their total phenolic, anthocyanin, and phenolic acid content. Overall, ‘Dinkum’ and ‘Josephine’ raspberry varieties possess higher total phenolic content, ORAC, DPPH radical scavenging activity, and α-glucosidase inhibitory activity than other five cultivars.     

Organic acids,antioxidant capacity,phenolic content and lipid characterisation of Georgia-grown underutilized fruit crops

Four underutilized Georgia-grown fruit crops, namely loquat (Eriobotrya japonica), mayhaw (Crataegus sp.), fig (Ficus carica), and pawpaw (Asimina triloba), and their leaves were analysed for total polyphenols by Folin–Ciocalteau method, and antioxidant capacity by ferric-reducing antioxidant power (FRAP) and Trolox-equivalent antioxidant capacity (TEAC) assays. Organic acids and phenolic compounds were identified by RP-HPLC. For lipid profile, fruits were separated into two fractions – seed and fruit (i.e., without seed); lipid was extracted using the Folch method and analysed for fatty acids, phytosterols, tocopherols, and phospholipids. The major organic acid identified in all samples was malic acid (177–1918 mg/100 g FW). The predominant phenolic acids in all the fruits were gallic (1.5–6.4 mg/100 g FW) and ellagic (0.2–33.8 mg/100 g FW), and the most abundant flavonoid was catechin (12.2–37.8 mg/100 g FW). Total lipid content varied from 0.1% in mayhaw fruit to 21.5% in pawpaw seed. Linoleic acid was the predominant fatty acid in all of the samples (28.2–55.7%).     

Microwave-assisted extraction of phenolics with maximal antioxidant activities in tomatoes

Microwave-assisted extraction (MAE) was investigated for extraction of phenolic compounds from tomato with maximised antioxidant activities using response surface methodology (RSM) coupled with a central composite design, and in vitro antioxidant assays (FRAP and ORAC). MAE was more efficient for greater antioxidant activities and higher total phenolic contents than solvent extraction. The optimal MAE processing parameters were 96.5 °C, 2.06 min, 66.2% ethanol for FRAP, and 96.5 °C, 1.66 min, 61.1% ethanol for ORAC. The models were successfully applied to 20 tomato cultivars, whose total phenolic contents (TPC) and indexes (TPI) were 489.30–997.45 mg gallic acid equivalent/100 g dry weight (DW) and 281.34–468.52 mg/100 g DW, respectively. Eight phenolic compounds were identified. Individual phenolics were 6.10–42.73 mg/100 g DW. The FRAP, but not the ORAC value showed good correlation with the TPC or TPI. The methodologies developed and the knowledge acquired in this study will provide useful information to tomato breeders and food processors.     

Nutritional composition of underutilized bayberry (Myrica rubra Sieb. et Zucc.) kernels

The kernels of five bayberry cultivars (Biqi, Zaodamei, Ding-ao, Dongkui and Wandao), grown in Zhejiang Province, China, were analyzed for their proximate composition, protein fractionation, amino acid profile, fatty acid composition and mineral contents. The antinutritive compounds, tannin and cyanide, were also quantified. These bayberry kernels possessed 25.0–27.64% DW protein. A majority of storage protein in bayberry kernels was of the globulin form. The kernel protein was rich in methionine, arginine, aspartic and glutamic acids while limiting amino acid was lysine. The most outstanding feature was the abundant fat content (62.5–68.1% DW) of kernels. Approximately 84.9–90.1% of the fatty acids were unsaturated with oleic acid (43.3–50.7%) and linoleic acid (34.1–46.8%). Bayberry kernels were good sources of magnesium, potassium and calcium. These results may offer a scientific basis for use of the under-exploited bayberry seeds.     

Changes of phenolic acids and antioxidant activities during potherb mustard (Brassica juncea, Coss.) pickling

Phenolic acids in potherb mustard (Brassica juncea, Coss.) were determined and the effects of pickling methods on the contents of total free phenolic acids, total phenolic acids, total phenolics, and antioxidant activities were investigated. Gallic acid, protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid were identified in the present study. The contents of total free phenolic acids, total phenolic acids and total phenolics in fresh potherb mustard were 84.8 ± 0.58 μg/g dry weight (DW), 539 ± 1.36 μg/g DW, and 7.95 ± 0.28 mg/g DW, respectively. The total free phenolic acids increased during the pickling processes, but the total phenolic acids, total phenolics, and antioxidant activities decreased. However, after 5 weeks of fermentation, all the pickling methods retained over 70% of total phenolic contents and above 65% of antioxidant capacities. The results indicated that pickling processes were relatively good methods for the preservation of phenolic acids and antioxidants for potherb mustard.     

The analysis of phenolic constituents in glabrous canaryseed groats

Nineteen glabrous canaryseed samples, comprising brown- and yellow-coloured seeds, were investigated to determine the nature of phenolic constituents present. Total phenolic content (TPC) was determined, using the Folin–Ciocalteau assay. Flavonoid and phenolic acid compositions were determined, using high performance liquid chromatographic and mass spectrometric (LC–MS/MS) techniques. TPC ranged from 174 to 209 mg/100 g for canaryseed wholemeal samples. The canaryseed bran contained twice as much TPC as the wholemeal. The brown- and yellow-coloured whole canaryseeds exhibited the same flavonoid profiles. LC–MS/MS analysis showed that the canaryseed acetone extract was rich in flavonoid glycosides, with the bran being mainly composed of O-pentosyl isovitexin and the flour having a compound at m/z 468. No proanthocyanidins were detected in the 19 samples. Ferulic acid was the dominant phenolic acid, followed by caffeic and p-coumaric acids. The wholemeal obtained from the brown-coloured group had significantly higher contents of ferulic (>196 mg/kg) and caffeic (>96 mg/kg) acids in comparison to the yellow-coloured canaryseed group. The latter had ferulic and caffeic acids at levels less than 165 and 78 mg/kg, respectively, with one exception which had relatively higher levels (190 and 94 mg/kg). Whilst canaryseed flour contained significantly very low levels of ferulic acid (22–34 mg/kg), the bran was enriched in ferulic (593–766 mg/kg), caffeic (304–452 mg/kg) and p-coumaric (119–142 mg/kg) acids.     

Identification and quantification of major phenolic compounds from mango (Mangifera indica, cv. Ataulfo) fruit by HPLC–DAD–MS/MS-ESI and their individual contribution to the antioxidant activity during ripening

Mango (Mangifera indica L.) is an economically important fruit throughout the world. ‘Ataulfo’ mango, a leading cultivar in Mexico, has the highest content of phenolic compounds among several commercial varieties of mango. However, the individual identification and antioxidant contribution of these phenols during ripening of mango fruit is unknown. Qualitative and quantitative analysis of the major phenolic compounds found in ‘Ataulfo’ mango fruit pulp was conducted in four stages of ripeness, using high-performance liquid chromatography coupled to mass spectrometry. The antioxidant contribution of each of the major phenolic compounds was calculated. The major compounds identified were chlorogenic acid (28–301 mg/100 g DW), gallic acid (94.6–98.7 mg/100 g DW), vanillic acid (16.9–24.4 mg/100 g DW), and protocatechuic acid (0.48–1.1 mg/100 g DW). The antioxidant contribution of the four phenolic acids increased during ripening. Gallic acid accounted for the highest contribution (39% maximum value), followed by chlorogenic acid (21% maximum value). This could indicate that these phenolic compounds may have an important role in the antioxidant metabolism in ‘Ataulfo’ mango fruit during ripening, and promoting health benefits to consumers.     

Antioxidant phenolic compounds loss during the fermentation of Chétoui olives

Evolution of phenolic compounds was studied during spontaneous and controlled fermentations of “Chétoui” cultivar olives at three degree of ripeness. Both spontaneous and controlled fermentations led to an important loss of total phenolic compounds with a reduction rate of 32–58%. Consequently, the antioxidant activity decreased by 50–72%. After fermentations, hydroxytyrosol and caffeic acid concentrations increased, whilst protocatechuic acid, ferulic acid and oleuropein concentrations decreased. The hydroxytyrosol concentration in black olives increased from 165 mg/100 g dry weight to 312 and 380 mg/100 g dry weight, respectively, after spontaneous and controlled fermentation. The oleuropein concentration in green olives decreased from 266 mg/100 g dry weight to 30.7 and 16.1 mg/100 g dry weight, respectively, after spontaneous and controlled fermentation. During olive fermentation, phenolic loss is essentially due to the diffusion of these compounds into the brine; the main phenolic compound identified and quantified in the different brines was hydroxytyrosol. To preserve antioxidant quality of table olives it is necessary to use a controlled process to minimise phenolic compound loss.     

In vitro antioxidant activities of three selected brown seaweeds of India

In vitro antioxidant activities of three selected Indian brown seaweeds – viz., Sargassum marginatum, Padina tetrastomatica and Turbinaria conoides – were investigated. Total phenolic content and reducing power of crude methanolic extract were also investigated. The activity of total methanolic extract and five different fractions (viz., petroleum ether (PE), ethyl acetate (EA), dichloromethane (DCM), butanol (BuOH) and aqueous) were studied using total antioxidant activity, DPPH radical scavenging and deoxyribose assays. EA fraction of S. marginatum exhibited higher total antioxidant activity of 39.62 mg ascorbic acid equivalent/g extract (or 0.31 mg ascorbic acid equivalent/g seaweed on dry weight basis) among the all the fractions. Among the fractions obtained from different seaweeds, EA fraction of S. marginatum showed higher DPPH scavenging activity of 23.16%; while PE fraction of T. conoides exhibited lower deoxyribose activity of 47.81%. Higher phenolic content (49.16 mg gallic acid equivalent/g extract or 0.86 mg GAE/g of seaweed on dry weight basis) was noticed in aqueous fraction of T. conoides. Reducing power of crude methanolic extract increased with increasing concentration. Reducing power of T. conoides and P. tetrastomatica were higher compared to standard antioxidant (α-tocopherol). Among the seaweeds, total methanolic extract of T. conoides had significantly higher phenol content (P < 0.05) compared to the other two species. In vitro antioxidant activity of methanolic extracts from all the three seaweeds showed an increase with increasing concentration indicating the dose dependency of these properties.     

Determination of free phenolic acids and antioxidant activity of methanolic extracts obtained from fruits and leaves of Chenopodium album

In this study, determination of phenolic acids as well as investigation of antioxidant activity of methanolic extracts from the fruits and leaves of Chenopodium album is described. Extracts were subjected to acidic hydrolysis in order to obtain total free phenolic acids. However, some of phenolic acids were identified and quantified by HPLC-DAD. The results were confirmed by LC-MS equipped with MS-ESI. In addition, Folin–Ciocalteu method was applied to determine the total phenolic contents. The antioxidant activity of C. album extracts was examined by using DPPH and hydroxyl radical-scavenging activity assays. Results revealed that the leaves extract exhibits better performance in antioxidant assays and in the higher total phenolic contents (3066 mg of GAE/100 g) when compared to fruits extract (1385 mg of GAE/100 g). From these results it has been revealed that the methanolic extracts of C. album from fruits and leaves have great potential as a source for natural health products.     

Microwave-assisted extraction of phenolic compounds from wine lees and spray-drying of the extract

Most research on extraction of phenol compounds from wine by-products and commercial exploitation of extracts use grape seeds and/or skins as raw materials. Looking for alternative antioxidants sources, obtaining antioxidant extracts from wine lees (also known as dregs), a sub-exploited by-product of winemaking process, is here presented. Microwave-assisted extraction (MAE) of phenolic compounds from wine lees has been optimized using the total phenols index, the ORAC values and yield of the extraction as response variables. Under the optimal working conditions, the proposed MAE method provides better extraction efficiency in a much shorter time (17 min) than the conventional extraction method for phenolic compounds (24 h). The liquid extract obtained by MAE was spray-dried. The type and amount of excipients used, as well as the spray-drying temperature, were optimized in order to minimize the oxidation of phenolic compounds and maximize the yield of the spray-drying process. The total phenols index in the dried extract thus obtained was 36.8% (expressed as gallic acid), showing an ORAC value of 3930 μmol TE/g. Additionally, Mv3G, Cm-Mv3G, myricetin, quercetin, quercetin-3-β-glucoside, caffeic acid and p-coumaric acid were quantified in the dry extract by HPLC–DAD. The results indicate that wine lees antioxidant extracts can be a suitable and cheap alternative to those obtained from grape seeds or skins.