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.     

Phenolics and antioxidant properties of bayberry (Myrica rubra Sieb. et Zucc.) pomace

Five cultivars of Myrica rubra, Biqi, Wandao, Dongkui, Dingao, and Zaodamei, were collected to analyze the phenolic compounds and evaluate the antioxidant properties of bayberry pomaces. The main anthocyanin was cyanidin-3-o-glucoside (3073.3–6219.2 mg/kg dry weight (DW)) and the main flavonol was quercetin-3-o-glucoside (296.2–907.9 mg/kg dry weight). Quercetin and myricetin were also found in the bayberry pomaces, and quercetin deoxyhexoside and myricetin deoxyhexoside were tentatively identified. The dominant phenolic acids were gallic acid (102.9–241.7 mg/kg dry weight) and protocatechuic acid (29.5–57.2 mg/kg dry weight). Other phenolic acids such as p-hydroxybenzoic, vanillic, caffeic, p-coumaric, and ferulic acids were also present in the bayberry pomaces, whereas, chlorogenic acid was only detected in Dongkui (1.58 mg/kg dry weight). The antioxidant activity of Wandao was the strongest of the five cultivars, whereas the activity of Dongkui was the weakest, and a significant positive relationship was observed between antioxidant activity and total phenolic content or total anthocyanins.     

Chemical characterization and antioxidant capacity of berries from Clidemia rubra (Aubl.) Mart. (Melastomataceae)

The flora of Latin America attracts gaining interest as it provides a plethora of still unexplored or under-utilized fruits that can contribute to human well-being due to their nutritional value and their content of bioactive compounds. Clidemia rubra (Aubl.) Mart. is a shrub belonging to the family of the Melastomataceae that grows preferably in a tropical climate. This paper comprises a nutritional characterization of the berries from Clidemia rubra and provides data on the phenolic compounds as well as the antioxidant capacity of the fruit. Findings in macronutrients like protein, carbohydrates, and fat were comparable to that of common berry fruits. Clidemia rubra berries seemed to be a good source for dietary fibers and some minerals (Ca, Mn, and Zn). In contrast, contents of titratable acids and ascorbic acid were low. The polyphenolic profile was determined by using HPLC-MS/MS in comparison to standard compounds. Noteworthy amounts of cyanidin 3-O-rutinoside (39.43 ± 1.66 mg/100 g fresh weight (FW)), delphinidin 3-O-rutinoside (23.74 ± 1.18 mg/100 g FW), cyanidin 3-O-glucoside (11.68 ± 0.56 mg/100 g FW), and delphinidin 3-O-glucoside (6.08 ± 0.35 mg/100 g FW) were found. Non-anthocyanin phenolic constituents were phenolic acids (gallic, protocatechuic, p-hydroxy-benzoic, vanillic, and caffeic acid), flavan-3-ols (epigallocatechin, epigallocatechin gallate, and epicatechin gallate), and 11 different myricetin and quercetin derivatives of which quercetin 3-O-arabinoside (5.26 ± 0.16 mg/100 g FW) and quercetin 3-O-rhamnoside (5.06 ± 0.08 mg/100 g FW) were dominating. Anthocyanins and ascorbic acid were mainly responsible for the antioxidant capacity of Clidemia rubra berries assessed with the total oxidant scavenging capacity (TOSC) assay.     

Polyphenol oxidase from bayberry (Myrica rubra Sieb. et Zucc.) and its role in anthocyanin degradation

Polyphenol oxidase (PPO) was extracted from bayberry (Myrica rubra Sieb. et Zucc. cv. Biqi), and its partial biochemical characteristics were studied. Stable and highly active PPO extracts were obtained using insoluble polyvinylpolypyrrolidone (PVPP) in sodium phosphate, pH 7.0, buffer. The highest PPO activity was observed in the ripe fruits. Optimum pH and temperature for bayberry PPO activity were pH 6.0 and T = 30 °C with 0.1 M catechol as substrate. PPO showed activity using the substrates of catechol, gallic acid and protocatechuic acid, but no activity with the substrates (+)-catechin, p-coumaric acid or caffeic acid. K_m and V_max values were 313 mM and 3.26ΔOD/min/g FW, respectively, with catechol as the substrate. Bayberry PPO did not act directly on cyanidin 3-glucoside but the rate of anthocyanin degradation was stimulated by the addition of gallic acid.     

Comparative antioxidant activity of extracts from leaves,bark and catkins of Salix aegyptiaca sp.

Leaves, bark and catkins of Salix aegyptiaca L. were extracted into solvents of increasing polarity from cyclohexane (non-polar), butanol, ethanol and water (polar) and analysed for their antioxidant capacity, total phenol and flavonoids. The highest antioxidant activity (19 μg/ml IC₅₀ for inhibition of DPPH radical activity), total phenolic content (212 mg gallic acid equivalents/g of dried extract) and total flavonoid (479 mg catechin equivalents/g of dried extract) was observed in the ethanolic extract of bark. HPLC identification of phenolic compounds from the extracts indicated the presence of gallic acid, caffeic acid, vanillin and p-coumaric acid, myricetin, catechin, epigallocatechin gallate, rutin, quercetin as well as salicin. Our data indicates the presence of high amounts of phenols and flavonoids in different parts of S. aegyptiaca species and propose that extracts from this plant may be utilised as a source of health promoting antioxidants.     

Phenolic profiles of in vivo and in vitro grown Coriandrum sativum L.

Coriandrum sativum L. is a source of a variety of polyphenols and other phytochemicals, related to its high antioxidant activity and to its use for indigestion, rheumatism, and prevention of lipid peroxidation damage. Plant cell cultures are a means to study or to produce some active metabolites, such as polyphenols. This technique was applied to the investigation of coriander, and a detailed analysis of individual polyphenols in vivo and in vitro grown samples was performed. The in vivo vegetative parts showed quercetin derivatives as the main flavonoids and quercetin-3-O-rutinoside (3296 mg/kg dw) was the main polyphenol found in this part of coriander. The fruits revealed only phenolic acids and derivatives, caffeoyl N-tryptophan hexoside (45.33 mg/kg dw) being the most abundant phenolic derivative. In vitro samples also gave a high diversity of polyphenols, being C-glycosylated apigenin (2983 mg/kg dw) the main compound. Anthocyanins were only found in clone A, which was certainly related to its purple pigmentation, and peonidin-3-O-feruloylglucoside-5-O-glucoside was the major anthocyanin found (1.70 μg/kg dw). In vitro culture can be used to explore new industrial, pharmaceutical, and medicinal potentialities, such as the production of secondary metabolites like flavonoids.     

Use of HPLC–DAD–ESI/MS to profile phenolic compounds in edible wild greens from Portugal

In some Mediterranean areas traditional wild greens are responsible for a significant percentage of total dietary antioxidant intakes. Asparagus acutifolius L. (wild asparagus), Bryonia dioica Jacq. (white bryony) and Tamus communis L. (black bryony) are important examples of those edible wild greens widely consumed. This study aimed to determine the phenolic profile and composition of edible vernal early shoots of those species. Wild asparagus and black bryony revealed glycosides of flavonols as the main phenolic compounds, while white bryony contained C-glycosylated flavones. Black bryony was the wild green that possessed the highest content of phenolic compounds (2200 mg/kg). Amongst the 11 flavonols found in this sample, kaempferol glycosides were the main compounds (1760 mg/kg). In the sample of wild asparagus, quercetin 3-O-rutinoside was the main flavonol found (263 mg/kg). Five flavones and one flavonol were found in the white bryony sample, apigenin 6-C-glucoside-7-O-glucoside being the major compound (1550 mg/kg).     

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%).     

Phenolic content and antioxidant capacity of tropical highland blackberry (Rubus adenotrichus Schltdl.) during three edible maturity stages

Tropical highland blackberry (Rubus adenotrichus Schltdl.) is a good source of antioxidants and contains appreciable levels of phenolic compounds, mainly ellagitannins and anthocyanins. This study examined the influence of three ripening stages on phenolic contents. Major anthocyanin pigments increased from 0.20 (red fruit) to 1.34 mg g⁻¹ fresh weight (FW) (fully ripe fruit), whereas ellagitannins and ellagic acid derivatives dropped from 3.8 to 2.2 mg ellagic acid equivalents g⁻¹ (FW). Flavonols also dropped from 5.1 to 2.0 mg quercetin equivalents 100 g⁻¹ (FW). Consequently, values for total phenolic compounds ranged from 5.8 to 5.2 mg gallic acid equivalents g⁻¹ (FW), showing no specific trend. Antioxidant activity (H-ORAC) increased from 38.29 to 64.00 μmol of Trolox equivalents g⁻¹ (FW) during ripening. When compared with other commercial cultivars, R. adenotrichus stands out for high H-ORAC value, although comparatively it possesses low anthocyanin content and average total phenolic content.     

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.     

Genotypic effects on the phytochemical quality of seeds and sprouts from commercial broccoli cultivars

Determining ontogenic diversity in vegetables, in general or Brassicas in particular, regarding the bioactive content, is an interesting task since the most promising Brassicaceae foods could have the highest nutritional and health-promoting properties. Therefore, vitamin C, phenolic compounds, glucosinolate content, and total antioxidant capacity were evaluated in commercial cultivars of broccoli (Brassica oleracea var. italica; cv. Nubia, cv. Marathon and cv. Viola) seeds and during the sprouting period. Vitamin C was not detected in dormant seeds and its content increased with the germination, reaching values ranging from 53 (cv. Nubia) to 64 (cv. Marathon) mg/100 g FW, at the end of the monitored period (14days). The total glucosinolate content in seeds and 3-day-old sprouts was higher in cv. Marathon (1005 and 556 mg/100 g FW, respectively), however cv. Viola sprouts registered the highest glucosinolate content 7 and 14 days after sowing (235 and 208 mg/100 g FW, respectively). Aliphatic glucosinolates were more affected by genetic factors than the indolic glucosinolates, being glucoraphanin the predominant glucosinolate in cv. Nubia and cv. Marathon, whereas glucoiberin was the major glucosinolate in cv. Viola. The flavonoid and total phenolic content was significantly higher in cv. Viola. Also, seeds of this cultivar showed the highest antioxidant capacity (2.7 mg Trolox/g FW).     

Polyphenol contents and in vitro antioxidant activities of lyophilised aqueous extract of kiwifruit (Actinidia deliciosa)

The aim of this study was to determine the antioxidant potency and total phenolic and flavonoid contents of kiwifruit (Actinidia deliciosa) in vitro by analysing the radical scavenging activity of lyophilised water extract from kiwifruit (LEK) for 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picryl-hydrazyl (DPPH), N,N-dimethyl-p-phenylenediamine (DMPD), and superoxide anion radical (O₂⁻) as well as the total reducing power by FRAP and CUPRAC assays and the metal chelating activities. LEK showed efficient radical scavenging activity with DPPH, ABTS, DMPD, and O₂⁻ radicals; ferric (Fe³⁺) and cupric (Cu²⁺) ion reducing power and metal chelating activities. Moreover, the amounts of phenolic compounds, such as caffeic acid, ferulic acid, syringic acid, ellagic acid, catechol, pyrogallol, p-hydroxy benzoic acid, vanillin, p-coumaric acid, gallic acid, quercetin, ??-tocopherol and ascorbic acid, in LEK were quantified by LC-MS-MS. The results show that pyrogallol (2070.0 mg/kg LEK) is the main phenolic compound responsible for the antioxidant and radical scavenging activities of LEK. Finally, total phenolic and flavonoid contents were determined as gallic acid (GAE) and quercetin equivalents (QE). The GAE and QE values in LEK were 16.67 ± 2.83 ??g GAE/mg and 12.95 ± 0.52 ??g QE/mg, respectively. The results suggest that consumption of kiwifruit (A. deliciosa) can be beneficial effects due to its antioxidant properties.     

Antioxidant and hepatoprotective activity of ethanolic extract of leaves of Solidago microglossa containing polyphenolic compounds

The antioxidative and hepatoprotective potential of Solidago microglossa D.C, a widely used medicinal plant from Brazil was investigated. The leaf extract showed inhibition against thiobarbituric acid reactive species (TBARS) induced by different prooxidants (10 μM FeSO₄ and 5 μM sodium nitroprusside SNP) in rat liver, brain and phospholipid homogenates from egg yolk. Moreover, the free radical scavenging activities of the extract was evaluated by the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) (IC_50, 3.8 ± 0.5 μg/ml) and hydroxyl radical on benzoic acid hydroxylation (IC_50, 32.3 ± 1.3 μg/ml) and deoxyribose (IC_50, 39.1 ± 2.4 μg/ml) assays. The ethanolic extract showed significant hepatoprotective activity against paracetamol (250 mg/kg) induced liver damage in mice in a dose dependent manner. The phenolic composition and their quantification by high performance liquid chromatography (HPLC) resulted in the identification of gallic acid and flavonoids: quercetrin (quercetin-3-O-rhamnoside), rutin (quercetin-3-O-rutinoside) and quercetin.     

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.     

The effects of solvents on the phenolic contents and antioxidant activity of Stypocaulon scoparium algae extracts

Water, water/methanol (1/1), methanol and ethanol crude extracts from a brown alga Stypocaulon scoparium were examined for total phenolic contents (TPC) using Folin–Ciocalteu method. DPPH scavenging assay was performed to measure the radical scavenging activities (RSA) of the extracts. Results showed a significant association between the antioxidant potency and the TPC. The aqueous extract showed both, the highest antioxidant activity and highest phenolic contents. The identification and quantification of phenolic antioxidants were carried out with a rapid and simple method of reverse phase high performance liquid chromatography (RP-HPLC). This method was developed for the simultaneous analysis of 14 polyphenols, namely gallic acid, catechin, epicatechin, rutin, p-coumaric acid, myricetin, quercetin and protocatechuic, vanillic, caffeic, ferulic, chlorogenic, syringic and gentisic acids. The chromatographic separation of 14 polyphenols was achieved in less than 40 min by RP-HPLC (Varian, Pursuit XRs C18 column, 5 μm, 250 mm × 4.6 mm) using linear gradient elution of methanol and water (0.1% formic acid) with a flow rate of 1 ml/min. Gallic acid was by far the predominant polyphenol.     

Phytochemical composition and antioxidant capacity of various botanical parts of the fruits of Prunus × domestica L. from the Lorraine region of Europe

Mirabelle plums represent a famous fruit from the Lorraine region, however little is known about their phytochemical composition. The oil of the fruit contained predominantly oleic acid (59%) and linoleic acid (29%). The total content of phenolic antioxidants in the whole fruits was 5.338 g/kg with 456 mg/kg (9%), 701 mg/kg (13%) and 4159 mg/kg (78%) detected in the peels, flesh and pits respectively. The peels contained solely 3,4-dihydroxybenzoic acid (270 mg/kg) and rutin (186 mg/kg), the flesh exclusively echinoids (723 mg/kg), whereas the pits contained a rich variety of phenolic compounds (4.2 g/kg) dominated by amygdalin (3.8 g/kg), but with significant contributions from vanillin (102 mg/kg), guajacyl-glycerin-coniferyl aldehyde isomers (87 mg/kg), dehydro-diconiferyl aldehyde (52 mg/kg), and vanillin diglucoside (48 mg/kg). Of the major phenolic compounds tested across a range of in vitro assays, rutin was the superior antioxidant.     

Biotransformation of phenolics (isoflavones,flavanols and phenolic acids) during the fermentation of cheonggukjang by Bacillus pumilus HY1

Changes in β-glycosidase, esterase activities, isoflavone, flavanols and phenolic acid during the fermentation of Korean whole soybean fermented food cheonggukjang by Bacillus pumilus HY1 were investigated. The levels of aglycones, flavanols and gallic acid increased, while the β-glucosidase activity, esterase activity, glycosides content (except for acetylglycosides) and flavanol gallates decreased. Total isoflavone content slightly decreased after 60 h of fermentation, while total flavanol and phenolic acid content increased. The highest levels of daidzein (aglycone type) and acetyldaidzin (glycoside type) were recorded after 48 h of fermentation. The levels of catechin, epicatechin and gallic acid also increased during fermentation. However, total contents of glycosides, malonylglycosides and flavanol gallates decreased by about 80%, 90% and 60% during 60 h of fermentation, respectively. In addition, total phenolic content increased markedly during fermentation, while levels of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity increased. Hence, it would be beneficial for the food industry if components of cheonggukjang could be separated and developed into functional products.     

Flavonoid constituents and their contribution to antioxidant activity in cultivars and hybrids of rabbiteye blueberry (Vaccinium ashei Reade)

Fruit from 42 blueberry (Vaccinium spp.) cultivars, including 36 rabbiteye cultivars (Vaccinium ashei Reade), three V. ashei hybrid derivatives, and three northern highbush (Vaccinium corymbosum L.) standards were evaluated for antioxidant capacity, individual flavonoid content, and the contribution of each identified phenolic compound to total antioxidant activity. Considerable variation was found in flavonoid content, antioxidant activity, and their contribution to total antioxidant activity among cultivars. Among 42 blueberry cultivars, the rabbiteye ‘Early May’ contained the highest amount of chlorogenic acid, myricetin 3-arabinoside, quercetin derivatives, and delphinidin-, cyanidin-, petunidin-, and malvidin-basis anthocyanins. ‘Early May’ cultivar also had the highest antioxidant activity (88.2 μmol TE/g fw). ‘Owen’, ‘Bluegem’, ‘Clara’, Climax’, and ‘Centurion’ were among the other rabbiteye cultivars that also had high levels of flavonoids and antioxidant activities. In contrast, the pink-fruited V. ashei hybrid, ‘Pink Lemonade’, had the lowest content of flavonoids and lowest antioxidant activity. The mean flavonoid content and antioxidant activity of rabbiteye cultivars was higher than those among northern highbush and V. ashei hybrids. The antioxidant activity of V. ashei hybrid derivatives was derived mainly from chlorogenic acid, myricetin, and quercetin, which contributed 62.5% of total antioxidant activity, whereas anthocyanins (malvidin, petunidin, delphinidin, and cyanidin) were the main contributors to the antioxidant activity of rabbiteye cultivars (76.2%) and northern highbush standards (76.8%). Blueberry cultivars identified to have high phenolic content and high antioxidant activity could be used as parents for future blueberry breeding programmes to develop new blueberry cultivars with higher antioxidant activity and further improve human health.