Characterisation of anthocyanins in the black soybean (Glycine max L.) by HPLC-DAD-ESI/MS analysis

The aim of this study was to isolate and identify the anthocyanins in the black seed coated soybean (cv. Cheongja 3, Glycine max (L.) Merr.) using reverse phase C-18 open column chromatography and high-performance liquid chromatography (HPLC) with diode array detection and electro spray ionization/mass spectrometry (DAD-ESI/MS) analysis, respectively. Anthocyanins were extracted from the coat of black soybeans with 1% TFA in methanol, isolated by RP-C-18 column chromatography, and their structures elucidated by 1D and 2D NMR spectroscopy. The isolated anthocyanins were characterised as delphinidin-3-O-glucoside (3), cyanidin-3-O-glucoside (5), petunidin-3-O-glucoside (6), pelargonidin-3-O-glucoside (7) and cyanidin (9). Furthermore, four minor anthocyanins were detected and identified as catechin-cyanidin-3-O-glucoside (1), delphinidin-3-O-galactoside (2), cyanidin-3-O-galactoside (4), and peonidin-3-O-glucoside (8) based on the fragmentation patterns of HPLC-DAD-ESI/MS analysis.     

Identification and characterisation of anthocyanins in the antioxidant activity-containing fraction of Liriope platyphylla fruits

The objective of this study was to investigate antioxidant activities and anthocyanin profiles in the fruits of Liriope platyphylla, where these are considered functional substances in Korea. The acidic methanol extract of this species exhibited potent antioxidant activities, showing 83.9% DPPH scavenging activity and 92.5% ABTS scavenging activity at a concentration of 0.5 mg/ml. Moreover, anthocyanins were identified by reversed-phase C₁₈ column chromatography, NMR spectroscopy, and HPLC-DAD-ESI/MS analysis. Seven anthocyanins were characterised, including delphinidin-3-O-glucoside (1), delphinidin-3-O-rutinoside (2), cyanidin-3-O-glucoside (3), petunidin-3-O-glucoside (4), petunidin-3-O-rutinoside (5), malvidin-3-O-glucoside (6), and malvidin-3-O-rutinoside (7). Among these, petunidin-3-O-rutinoside (5) (7302.2 μg/g) and malvidin-3-O-rutinoside (7) (5776.1 μg/g) were the predominant anthocyanins, whereas the least prevalent anthocyanin was found to be cyanidin-3-O-glucoside (3) (64.9 μg/g). Therefore, our results suggest that strong antioxidant activities of the acidic methanol extract of L. platyphylla fruits are correlated with high anthocyanin contents, particularly the petunidin-3-O-rutinoside (5) and malvidin-3-O-rutinoside (7).     

云南丽江和吉林靖宇11种越橘果实花色苷组分的分析

采用高效液相色谱法对云南丽江、吉林靖宇11种越橘果实花色苷组分进行测定。通过标准品的分析,建立了飞燕草素-3-半乳糖苷、飞燕草素-3-葡萄糖苷、矢车菊素-3-半乳糖苷、矢车菊素-3-葡萄糖苷、锦葵色素-3-半乳糖苷、锦葵色素-3-葡萄糖苷的回归方程,相关系数为0.9939~0.9968,精密度RSD为2.00%~3.73%,回收率在98.90%~100.99%,方法准确可行。通过6种标准品的对比,供试的15个样品中,所有越橘品种均有飞燕草素-3-半乳糖苷、矢车菊素-3-半乳糖苷、锦葵色素-3-半乳糖苷3种花色苷,飞燕草素-3-半乳糖苷平均含量为223.99 μg/g、锦葵色素-3-半乳糖苷平均含量为153.34 μg/g,二者占总花色苷的65%。采用欧氏距离聚类分析表明,集群1为飞燕草素-3-半乳糖苷含量较高的三种越橘,代表品种为丽江雷戈西、丽江奥尼尔、靖宇杜克;集群2为6种花色苷总含量较高的越橘品种,代表品种为丽江北陆与靖宇早蓝,说明飞燕草素-3-半乳糖苷含量与花色苷总含量是评价越橘花色苷特点的重要因子。     

Identification of anthocyanins and distribution of flavonoids in tamarillo fruit (Cyphomandra betaceae (Cav.) Sendt.)

The major tamarillo (Cyphomandra betaceae) anthocyanin pigments were isolated and identified as pelargonidin-3-rutinoside, pelargonidin-3-glucoside, cyanidin-3-rutinoside, cyanidin-3-glucoside, delphinidin-3-rutinoside and delphinidin-3-glucoside. The intense purple-coloured jelly surrounding the seeds contained the greatest concentration of anthocyanins, delphinidin-3-rutinoside being the major pigment. Flavones, flavonols and leucoanthocyanins were also present in this material. The yellow-coloured flesh contained flavones and low concentration of anthocyanins. The major anthocyanin of the skins is cyanidin-3-rutinoside; flavones and leucoanthocyanins are also present. It is suggested that the presence of leucoanthocyanins in pigment extracts induced degradation of anthocyanins during isolation and purification.     

Phenolic Compounds from the Leaves of Vitis labrusca and Vitis vinifera L. as a Source of Waste Byproducts: Development and Validation of LC Method and Antichemotactic Activity

The cultivation of Vitis (Vitaceae) grape varieties is one of the most important economic activities in agribusiness in southern Brazil. Vitis varieties are rich in polyphenolic compounds with several pharmacological and biological activities, such as antioxidant action. In this context, this study analyzed qualitatively and quantitatively the anthocyans and flavonoids found in the leaves of grape varieties Vitis vinifera and Vitis labrusca. For this purpose, vine leaf extracts were prepared and the chemical profile of each was characterized by LC/MS-MS. Two high performance liquid chromatography-validated methods were performed using UV/VIS-LC-DAD detector to quantify phenolic compounds. The main anthocyanins isolated from vine leaves were cyanidin-3-O-glucoside and peonidin-3-O-glucoside. The flavonoids identified were rutin, quercetin-3-O-galactoside, quercetin-3-O-glucoside, and quercetin-3-O-glucuronide, which was the predominant compound. The Waters X-Terra® RP18 column allowed the effective separation of quercetin-3-O-glucuronide from the other flavonoids for the first time, besides the partial separation of quercetin-3-O-galactoside from quercetin-3-O-glucoside. Furthermore, another phenolic compound was confirmed by MS spectrometry, using direct infusion, as being trans-caftaric acid. The present study also investigates the antichemotactic activity in vitro of grape crude extracts, fractions, and isolated compounds. It was demonstrated that almost all fractions and isolated compounds showed increased antichemotactic effect in response to LPS with a more pronounced values of IC₅₀ for anthocyanins fraction, rutin, quercetin-3-O-galactoside, and trans-caftaric acid (0.9, 1.6, 3.7, and 5.1 ng/mL, respectively).     

Identification and quantification of anthocyanins from the grains of black rice (Oryza sativa L.) varieties

The purpose of this research was conducted to determine anthocyanin content from the grains of 10 Korean black rice varieties. Moreover, the primary constituents including protein and oil were evaluated. Anthocyanins, cyanidin-3-O-glucoside (1) and peonidin-3-O-glucoside (2), were isolated and elucidated using reversephase C₁₈ chromatography, nuclear magnetic resonance (NMR) spectroscopy, and high performance liquid chromatography (HPLC) with diode array detection and electrospray ionization/mass spectrometry (DAD-ESI/MS). Anthocyanin showed significant differences and cyanidin-3-O-glucoside (1) (52.1±6.3-1,601.0±8.5 μg/g) exhibited a markedly higher content than peonidin-3-O-glucoside (2) (ND-82.6±1.2 μg/g). Among varieties, ‘Heugjinjubyeo’ showed the highest anthocyanin (1: 1,601.0±8.5, 2: 82.6±1.2 μg/g), whereas ‘Heughyang’ was the lowest (1: 52.1±6.3 μg/g, 2: ND). Protein and oil exhibited the minor differences and their contents ranged from 10.7±0.0 to 14.1±0.1% and from 2.1±0.0 to 2.9±0.0%. Overall results suggest that anthocyanin can be a key factor in functional property of black rice and ‘Heugjinjubyeo’ may be very important source concerning nutritional value.     

Characterisation of phenolic phytochemicals and quality changes related to the harvest times from the leaves of Korean purple perilla (Perilla frutescens)

The objective of this study was to investigate the profiles of phenolic phytochemicals in the leaves of Korean purple perilla (cv. Bora, Perilla fructescens) using reversed-phase C₁₈ column chromatography and HPLC with DAD-ESI/MS analysis. Changes in their contents were also the first reported through eight different harvest times during two months. They were characterised as five anthocyanins and three phenolic acids including cyanidin-3,5-di-O-glucoside (1), cyanidin-3-O-glucoside (2), cyanidin-3-O-(6-O-caffeoyl)glucoside-5-O-glucoside (3), cyanidin-3-O-(6-O-coumaroyl)glucoside-5-O-glucoside (4), cyanidin-3-O-(6-O-coumaroyl)glucoside (5), caffeic acid (6), rosmarinic acid (7), and rosmarinic acid methylester (8). Significant differences were observed between individual and total phytochemical contents, especially, cyanidin-3-O-(6-O-coumaroyl)glucoside-5-O-glucoside (4) and rosmarinic acid (7) exhibited the predominant constituents. Among different harvest times, the highest content was found with 82.473 mg/g on 21st September, while the lowest was 39.000 mg/g on 17th August. These results may be useful in determining the optimal harvest time at which phenolic phytochemicals reaches a maximum level in mid-September.     

Identification of phenolic compounds isolated from pigmented rices and their aldose reductase inhibitory activities

Two anthocyanins (cyanidin-3-O-β-glucoside and peonidin-3-O-β-glucoside) and other phenolic (ferulic acid) were, respectively isolated from black and pigmented brown rices (Oryza sativa L. japonica) and their complete structures were determined by spectroscopic analyses (H NMR, C NMR and MALDI MASS). The HPLC profile of anthocyanins extracted from black rice showed cyanidin-3-O-β-glucoside as the first peak (85%) and peonidin 3-O-β-d-glucoside as the second (15%), while that of pigmented brown rice showed ferulic acid as the first peak (85.7%) and tocols as the second (14.3%). Several tocols were isolated and identified from the unsaponifiable fractions of both rices having some difference on their structures and amounts. The aldose reductase inhibitory activity of isolated compounds was in the following decreasing order: cyanidin-3-glucoside > quercetin > ferulic acid > peonidin-3-glucoside > tocopherol.     

Chromatographic Separation of Anthocyanins in Cowberry (Lingonberry) Vaccinium vites-idaea L

The analysis of anthocyanins by a combination of droplet counter-current chromatography, thin-layer chromatography and on-line high-performance liquid chromatography was applied to the acidified ethanol extract of cowberry (lingonberry), Vaccinium vitesidaea L, In addition to 3-galactoside (88.0%), 3-arabinoside (10.6%), and 3-glucoside (1.4%) of cyanidin, delphinidin-3-glucoside (less than 0.1%) was identified. The total anthocyanin content was determined to be 174 mg/100g fresh fruit.     

Pattern recognition of three Vitis vinifera L. red grapes varieties based on anthocyanin and flavonol profiles,with correlations between their biosynthesis pathways

The presence of anthocyanins and flavonols in three selected red grape varieties was investigated, in order to use their polyphenolic characterisation as a fingerprint. Berry skins of Gran Negro grapes were characterised by the presence of high content of malvidin- and peonidin-3-O-glucoside; Mouratón grapes, by the presence of high content of petunidin- and delphinidin-3-O-glucoside; and Brancellao grapes, by the presence of high content of cyanidin-3-O-glucoside. The main flavonols found included the 3-O-glucosides of quercetin, myricetin, kaempferol, laricitrin, isorhamnetin and syringetin. Using cluster analysis and principal components analysis, Gran Negro could be characterised by their content of isorhamnetin-3-O-glucoside and syringetin-3-O-glucoside and, along with Mouratón, by their myricetin conjugates. Flavonol profile could not provide a fingerprint of Brancellao variety. Stepwise discriminant analysis was performed in order to find the polyphenolic compounds, which characterised the selected grape varieties. Finally, anthocyanin and flavonol profiles in red grapes were compared and results confirmed that biosynthesis of flavonols is closely related to that of anthocyanins.     

Phenolic Analyses of Haskap Berries (Lonicera caerulea L.): Spectrophotometry Versus High Performance Liquid Chromatography

Haskap berries (Lonicera caerulea L.) are known for their high phenolics, anthocyanins, and antioxidant potential. The data on the phenolic profile of these fruits are lacking. In this study, the phenolic profiles of three haskap varieties; tundra, berry blue, and indigo gem grown in Nova Scotia, Canada were investigated for the first time using spectrophotometery and high-performance liquid chromatography. Berries were analyzed for total phenolic content/total reducing capacity, total anthocyanin content, and antioxidant potential (2,2-diphenyl-1-picrylhydrazyl radical scavenging activity). The total reducing capacity, total anthocyanin content, and 2,2-diphenyl-1-picrylhydrazyl values were 6.17–8.42 mg gallic acid equivalents/gram fresh weight, 4.49–6.97 mg cyanidin-3-glucoside equivalents/gram fresh weight, and 78.70–89.55%, respectively. The extracts were analyzed by reversed-phase diode array detector–high-performance liquid chromatography through a gradient elution using Synergi 4 µm Max-RP C12 column and the chromatograms were acquired at 520, 360, and 320 nm for athocyanins, flavonoids, and free phenolic acids, respectively. The identified anthocyanins were cyanidin-3-glucoside (82.81–91.99% of the total anthocyanins), cyanidin 3,5-di-glucoside (2.31–4.27%), cyanidin-3-rutinoside (1.54–9.20%), peonidin-3-O-glucoside (0.75–3.44%), and pelargonidin-3-glucoside (0.77–2.98%). Other flavonoids (quercetin-3-β-D-glucoside and quercetin-3-rutinoside) and free phenolic acids (chlorogenic and neochlorogenic) were also quantified.     

龙葵果花色苷的分离与鉴定

采用硅胶柱层析技术分离制备龙葵果花色苷。分离得到的2 个花色苷馏分经紫外-可见光谱、高效液相色谱-电喷雾串联质谱（high performance liquid chromatography electrosprary ionization-mass spectrometry,HPLC-DADESI-MS/MS）进行结构鉴定,并结合酸水解分析糖苷种类。最终确定馏分Ⅰ为飞燕草素-3-琥珀酰阿拉伯糖苷,根据峰面积归一化法计算其纯度为94%;馏分Ⅱ为矢车菊素-3-半乳糖苷和矢车菊素-3-乙酰半乳糖苷,峰面积归一化法计算其纯度分别为45.67%和13.97%。     

Changes occurring in compositional components of black soybeans maintained at room temperature for different storage periods

Changes in compositional components of black soybeans, including isoflavone, anthocyanin, protein, oil, and fatty acid, were investigated for the first time in soybeans maintained at room temperature for different storage periods. Isoflavone and anthocyanin profiles in hydrolysed extracts were characterised by column chromatography and HPLC-DAD-ESI/MS spectrometry analysis. These components decreased markedly during storage, whereas protein, oil, and fatty acid showed a slight decrease. The individual isoflavones and anthocyanins observed in black soybeans were as follows, in order of abundance: genistein > daidzein > glycitein; cyanidin-3-O-glucoside > dephinidin-3-O-glucoside > petunidin-3-O-glucoside. In particular, genistein (518.4 → 415.7 → 274.8 μg/g) and cyanidin-3-O-glucoside (6.53 → 2.92 → 1.49 mg/g) showed the greatest decrease for a storage time of two years. The scavenging activities of DPPH and ABTS radicals during storage also decreased in comparison with those of observed before storage. Our results can be used to improve our understanding of the relationship between storage times and the components from black soybeans.     

Characterisation and preliminary bioactivity determination of Berberis boliviana Lechler fruit anthocyanins

Berberis boliviana Lechler is a member of the Berberidaceae family that has a small edible red-purple berry. The plant is native to the Peruvian Andes and contains high amounts of anthocyanin pigments. The monomeric anthocyanin content, determined by a pH-differential method, was 7/100 g of seedless berries. Pigments were characterised by HPLC coupled to a photodiode array (PDA) and mass spectrophotometer (MS) detectors. Five aglycones and ten anthocyanins were found and identified as petunidin-3-glucoside (24.4%), delphinidin-3-glucoside (24.1%), malvidin-3-glucoside (22.1%), cyanidin-3-glucoside (10.2%), petunidin-3-rutinoside (7.15%), malvidin-3-rutinoside (4.9%), cyanidin-3-rutinoside (3.8%), delphinidin-3-rutinoside (2.6%), peonidin-3-glucoside (1.1%), and peonidin-3-rutinoside (0.9%).     

Flavonoids, sugars and fruit acids of alpine bearberry (Arctostaphylos alpina) from Finnish Lapland

Alpine bearberry (Arctostaphylos alpina L.) is a special circumpolar edible berry primarily used for household purposes but with greater potential in commercial applications than utilized thus far. In this study, the flavonoids of alpine bearberry were investigated with HPLC-DAD and HPLC-MS, and the sugars and fruit acids as trimethylsilyl derivates with GC-FID. The most abundant anthocyanin was cyanidin-3-O-galactoside accounting for over 95% of the anthocyanins. Eight other anthocyanins existed in trace amounts only, the richest of them being cyanidin-3-O-arabinoside and cyanidin-3-O-glucoside. Cyanidin accounted for over 99% of the anthocyanin aglycones. All the flavonols identified were glycosides of quercetin. Majority of the sugars were glucose and fructose while the most abundant fruit acid was quinic acid. The unique anthocyanin composition of alpine bearberry encourages advanced use of the berry as a food ingredient in private households and in industrial applications. In addition, it serves as a unique reference material for research purposes.     

Influence of Dekkera bruxellensis on the contents of anthocyanins,organic acids and volatile phenols of Dão red wine

The anthocyanin, organic acid and volatile phenol compositions of red wine obtained from Touriga Nacional grapes growing in the Dão region (Portugal) were determined by HPLC/DAD, HPLC/UV and GC/FID, respectively. By these means, nine anthocyanic compounds (malvidin-3,5-O-diglucoside, cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, peonidin-3-O-glucoside, malvidin-3-O-glucoside, delphinidin, cyanidin, pelargonidin and malvidin), six organic acids (ketoglutaric, tartaric, malic, quinic, lactic and shikimic acids) and two volatile phenols (4-ethylguaiacol and 4-ethylphenol) were identified and quantified. Malvidin-3-O-glucoside, the pair lactic plus shikimic acids and 4-ethylguaiacol were the main anthocyanin, organic acids and volatile phenol, respectively. The effects of nine different Dekkera bruxellensis strains on these chemical parameters were also evaluated. The results obtained indicate that some strains of D. bruxellensis yeast are able to cause deterioration of red wine from the Dão region during its maturation by the production of volatile phenols, namely 4-ethylphenol.     

Phenolic compounds in natural black Spanish olive varieties

There is increasing interest in olive phenolic compounds because of their biological properties as well as their contribution to the colour, taste and shelf life of olive products. Phenolic compounds in natural black Spanish olives are characterised by HPLC-UV and HPLC-MS. Hydroxytyrosol-4-#-D-glucoside has been identified as the major phenolic compound in natural black olives. A qualitative and quantitative study of phenolic substances such as hydroxytyrosol, salidroside, tyrosol, verbascoside, luteolin-7-glucoside, rutin, demethyloleuropein and oleuropein has been carried out. Traces of other phenolic compounds such as catechol, vanillic acid, apigenin-7-rutinoside and ligustroside have also been found by mass spectroscopy. Secoiridoid aglycons, the main phenolic compounds in olive oil, were not detected in olive flesh except the dialdehydic form of elenolic acid linked to hydroxytyrosol (HyEDA), particularly in the Arbequina olive variety and only traces were detected in other varieties. Among anthocyanins, cyanidin-3-O-glucoside and cyanidin-3-O-rutinoside were the only anthocyanin compounds detected in all the varieties studied. The UV and MS spectra of several unknown phenolic compounds have also been described.     

Changes of flavonoid content and antioxidant capacity in blueberries after illumination with UV-C

The levels of flavonoids in blueberries (Vaccinium corymbosum L.) were found to increase after illumination with UV-C. Phytochemicals affected included resveratrol, myricetin-3-arabinoside, quercetin-3-galactoside, quercetin-3-glucoside, kaempferol-3-glucuronide, delphinidin-3-galactoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, petunidin-3-galactoside, petunidin-3-glucoside, petunidin-3-arabinoside, malvidin-3-galactoside, malvidin-3-arabinoside, and chlorogenic acid as analyzed by HPLC. Significantly higher antioxidant capacity was detected in fruit treated with 2.15, 4.30, or 6.45 kJ m⁻² compared to the control fruit. UV-C dosage of 0.43 kJ m⁻² also increased phenolics and anthocyanins, but to a lesser extent. The optimum doses of UV-C for enhancing phytochemical content in blueberries were 2.15 and 4.30 kJ m⁻². These data suggest that proper use of UV-C illumination is capable of modifying the phytochemical content of blueberries. Time course measurements of the effects of UV-C revealed that the strongest responses of fruit to UV-C treatment occurred instantly after the illumination and the effects diminished with time. Therefore, even though residual effects were evident following UV-C exposure, the best results were obtained immediately after the treatment.     

Soluble phenolics and antioxidant properties of soybean (Glycine max L.) cultivars with varying seed coat colours

This research was the first to investigate nutritional components, including soluble phenolics (isoflavones and anthocyanins), protein, oil, and fatty acid as well as antioxidant activities in different coloured seed coat soybeans (yellow, black, brown, and green) for two crop years. The soluble phenolics differed significantly with cultivars, crop years, and seed coat colours, while protein, oil, and fatty acid exhibited only slight variations. Especially, malonylgenistin and cyanidin-3-O-glucoside compositions had the most remarkable variations. Green soybeans had the highest average isoflavone content (3079.42 μg/g), followed by yellow (2393.41 μg/g), and black soybeans (2373.97 μg/g), with brown soybeans showing the lowest value (1821.82 μg/g). Anthocyanins showed only in black soybeans, with the average contents of the primary anthocyanins, cyanidin-3-O-glucoside, delphinidine-3-O-glucoside, and petunidin-3-O-glucoside, quantified at 11.046, 1.971, and 0.557 mg/g, respectively. Additionally, Nogchae of green soybean and Geomjeongkong 2 of black soybean may be recommended as potential cultivars owing to the highest average isoflavone (4411.10 μg/g) and anthocyanin (21.537 mg/g) contents. The scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals also differed remarkably, depending upon isoflavone and anthocyanin contents, with black soybeans exhibiting the highest antioxidant effects.     

Kinetics of Thermal Degradation of the Anthocyanins Delphinidin-3-rutinoside and Malvidin-3-glucoside

The kinetics of the thermal degradation of the two anthocyanins delphinidin-3-rutinoside and malvidin-3-glucoside at 78, 88, 98 and 108 °C, depending on the pH and the type of solvent has been investigated. The first order reaction of delphinidin-3-rutinoside and malvidin-3-glucoside degradation has been established. The rate constants (K ± 10^?5s^?1), the activation parameters, factor Q₁₀ and the half-decay period have been calculated. The rate of delphinidin-3-rutinoside degradation is higher than that of malvidin-3-glucoside.