Characterization and quantification of saponins and flavonoids in sprouts,seed coats and cotyledons of germinated black beans

Saponins, flavonols and isoflavones were quantified in sprouts, cotyledons and seed coats of black beans (Phaseolus vulgaris L.) subjected to germination over five days. Sprouts had a higher concentration of saponins compared to cotyledons or seed coats (p < 0.05). The saponins concentration in hilum increased 2.3-fold after soaking. After the first day of germination, the saponin concentration in sprouts and cotyledons increased 1.9 and 2.1-fold, respectively. Additional germination days decreased the amount of the most abundant soyasaponins in black bean sprouts. Flavonols and isoflavones were associated with seed coats and less than one third of the initial amount remained after the soaking process. The concentrations of flavonols were also reduced during germination process. Aglycones were detected only after soaking and their concentration remained unchanged during germination. Genistein was detected only after three days of germination. In general, one-day germinated black beans could be recommended for increasing the concentration of saponins and non-glycosylated flavonols in sprouts and seed coats, respectively.     

Comparisons of phenolic compounds, isoflavones, antioxidant capacity and oxidative enzymes in yellow and black soybeans seed coat and dehulled bean

The total and individual phenolic profiles and their contribution to antioxidant capacities of seed coats and dehulled beans of black and yellow soybeans were investigated. In addition, lipoxygenase (LOX) and peroxidase (POD) activities were also determined. Black seed coats had the highest content of total phenols, flavonoids and anthocyanins, in addition the highest catechin and protocatechin contents. On the other hand, dehulled beans from yellow soybeans had the highest levels of ferulic and p-coumaric acid content, and free and bound isoflavone content. The LOX activity was concentrated in the dehulled beans of both black and yellow soybeans, while a high POD activity that can be attributed to a dominant gene EpEp was presented in seed coats of yellow soybean genotype Galeb. Due to the high concentration of phenols, the black seed coats from soybean genotypes Black Tokio and Cornaja had the highest ABTS^·+-scavenging capacity (597.46 and 486.15 mmol Trolox Eq./kg d.m., respectively). The results suggest that the black seed coat as well as dehulled bean from yellow soybeans would potentially provide sources of natural antioxidants that may play a crucial role in human health protection.     

Effect of cooking and germination on phenolic composition and biological properties of dark beans (Phaseolus vulgaris L.)

Legumes are the base´s diet in several countries. They hold a high nutritional value, but other properties related to human health are nowadays being studied. The aim of this work was to study the influence of processes (boiling or germination) on the phenolic composition of dark beans (Phaseolus vulgaris L. c.v. Tolosana) and their effect on their antioxidant, neuroprotective and anticancer ability. Phenolic composition of raw and processed dark beans was analysed by HPLC-PAD and HPLC–ESI/MS. The antioxidant activity was evaluated by ORAC. Astrocytes cultures (U-373) have been used to test their neuroprotective effect. Anticancer activities were evaluated on three different cell lines (renal adenocarcinoma (TK-10), breast adenocarcinoma (MCF-7) and melanoma (UACC-62)) by sulphorhodamine B method. Qualitative and quantitative differences in phenolic composition have been observed between raw and processed dark beans that influence the antioxidant activity, mainly for germinated samples which show a decrease of antioxidant capacity. Although every assayed extracts decreased reactive oxygen species release and exhibited cytotoxicity activities on cancer cell lines, raw beans proved to be the most active in neuroprotective and antitumoral effects; this sample is especially rich in phenolic compounds, mainly anthocyanins. This study further demonstrated that phenolic composition of dark beans is related with cooking process and so with their neuroprotective and anticancer activity; cooking of dark beans improves their digestion and absorption at intestinal level, while maintaining its protective ability on oxidative process at cellular level.     

Elucidation of the flavonoid and furocoumarin composition and radical-scavenging activity of green and ripe chinotto (Citrus myrtifolia Raf.) fruit tissues,leaves and seeds

Citrus myrtifolia Raf. (chinotto) flavedo, albedo and carpel membranes from green and ripe fruits, as well as seeds and leaves, were investigated for the first time for their flavonoid and furocoumarin composition. Twenty-three unique compounds distributed in the different fruit/plant parts were identified and quantified. All samples analysed contained flavanone O-glycosides, flavone C- and O-glycosides and furocoumarins; flavedos and albedos also contained significant amounts of polymethoxyflavones. Flavedo and albedo extracts were found to be richest in flavonoids and furocoumarins, containing up to 1.95 g/kg fresh weight. Flavedo, albedo and carpel membranes from ripe fruits were found to be richer than the corresponding tissues from unripe fruits. The remarkable radical-scavenging activity of all the extracts was tested by DPPH^•, ABTS^•+ and FRAP methods, revealing that (i) they were particularly efficient in quenching ABTS^•+ radical cations (up to 9.8 mM Trolox equivalents), and (ii) flavedo and albedo extracts, on average, showed the highest antioxidant capacity.     

Modulation of the antioxidant/pro-oxidant balance,cytotoxicity and antiviral actions of grape seed extracts

Grape seed extracts (GSEs) were investigated in yeast cells harbouring defects in their antioxidant system (regarding the cellular growth and growth recovery from H₂O₂ insult). GSEs antioxidant activity was detected in wild-type and mutant strains Δcta1, Δgsh1 and Δoye2glr1, while pro-oxidant activity in Δsod1 cells was seen. Assessment of proliferation of prostate cancer PC3 and HBV-replicating HepG2 2.2.15 cells treated with GSEs has shown higher cytotoxicity of red grape seed extract (RW) than white grape seed extract (WW) subjective to dose and period of administration. No antiviral effect was detected by measuring the secreted virion particles in HepG2 2.2.15 cells treated with GSEs. The GSEs play a dual antioxidant/pro-oxidant role in vivo according with the cellular antioxidant system deficiencies and exhibit cytotoxic properties in PC3 and HepG2 2.2.15 cell lines, but no antiviral action against HBV.     

Flavonoid content and antioxidant activity of vegetables from Indonesia

Extracts from 11 vegetables of Indonesian origin were screened for flavonoid content, total phenolics, and antioxidant activity. The flavonols myricetin, quercetin, and kaempferol and flavones luteolin and apigenin were quantified by HPLC. Flavonoid content in mg/100 g fresh weight (fw) was apparently initially reported for Cosmos caudatus H.B.K. (52.19), Polyscias pinnata (52.19), Pluchea indica Less. (6.39), Nothopanax scutellarius (Burm.f.) Merr (5.43), Talinum triangulare (Jacq.) Willd. (3.93), Pilea melastomoides (Poir.) Bl. (2.27), and Etlingera elatior (Jack) R.M.Sm (1.18). The flavonoid content of the vegetables studied were mainly quercetin and kaempferol and ranged from 0.3 to 143 mg/100 g fw, with the highest level found in Sauropus androgynus (L) Merr. C. caudatus H.B.K. had the greatest total phenols among the vegetables analysed, with 1.52 mg GAE/100 g fw. P. indica Less. and C. caudatus H.B.K. had the highest antioxidant activity as measured by ferric cyanide reducing power, DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) scavenging, and inhibition of linoleic acid oxidation. Therefore, S. androgynus (L) Merr, C. caudatus H.B.K., and P. pinnata were identified as potentially rich sources of dietary flavonoids and antioxidants.     

The antioxidant properties of the cherimoya (Annona cherimola) fruit

Annona cherimola is an exotic fruit from the genus Annona, native to the Andean highlands in western South America. The cherimoya skin, flesh and juice were isolated and analyzed for antioxidant content using the oxygen radical absorbance capacity (ORAC) assay. The juice showed the highest antioxidant activity, while the flesh exhibited the lowest. The cherimoya skin, flesh and juice extracts were also incubated with Raji (Burkitt's Lymphoma) and HT-29 (colon cancer) cell lines, and the antioxidant uptake of cells was measured. Both cell lines, when subjected to cherimoya juice, showed the highest antioxidant uptake. The cells were then exposed to AAPH, a radical initiator, to simulate the conditions of cells under oxidative stress, and then subjected to cherimoya skin, flesh and juice extracts. Both cell lines absorbed more antioxidants after being pre-exposed to AAPH, indicating that cells under stress have the ability to import antioxidants.     

Biological activities of flavonoids from Nitraria retusa (Forssk.) Asch. and their acylated derivatives

The antioxidant activity and antiproliferative property towards Caco2 cells of water and methanol extracts/fractions of Nitraria retusa were investigated. The total phenolic and flavonoid contents of the extracts/fractions were determined, and the four major flavonoids were identified as isorhamnetin, isorhamnetin-3-O-glucoside, isorhamnetin-3-O-rutinoside and isorhamnetin-3-O-robinobioside. The results showed a relationship between the extracts/fractions activities and their flavonoid contents. Moreover, the chloroform extract which was enriched with the aglycone flavonoid isorhamnetin exhibited the highest activities. The activities of N. retusa flavonoids were compared to those of model flavonoids, quercetin, isoquercitrin and rutin. In all cases, the aglycone compounds were more active than their glycosylated derivatives. Isorhamnetin-based flavonoids presented higher antiproliferative activities than quercetin-based ones, while similar antioxidant properties were observed. The enzymatic acylation of isorhamnetin-3-O-glucoside with ethyl laurate and ethyl butyrate enhanced its capacity to inhibit xanthine oxidase and its antiproliferative activity but decreased its radical scavenging activity.     

Reduction of antiproliferative capacities,cell-based antioxidant capacities and phytochemical contents of common beans and soybeans upon thermal processing

The effects of boiling and steaming processes on the antiproliferative and cellular antioxidant properties, as well as phytochemicals, of two types of common beans (pinto and black beans) and two types of soybeans (yellow and black) were investigated. All thermal-processing methods caused significant (p < 0.05) decreases in total phenolic content (TPC), total saponin content (TSC) and phytic acid content (PAC) values in all bean types (except for TPC values in pressure-steamed yellow soybeans) as compared to those of the raw beans. All types of uncooked raw beans exhibited cellular antioxidant activities (CAA) in dose-dependent manners. Black soybeans exhibited the greatest CAA, followed by black beans, pinto beans and yellow soybeans. The CAA of cooked beans were generally diminished or eliminated by thermal processing. The hydrophilic extracts from raw pinto beans, black beans and black soybeans exhibited antiproliferation capacities against human gastric (AGS) and colorectal (SW480) cancer cells in dose-dependent manners. The raw yellow soybeans exhibited dose-dependent antiproliferation activities against the SW480 cells. Most of the cooked beans lost their antiproliferation capacities as observed in the raw beans. These results indicate that different processing methods may have various effects on phytochemical profiles and bioactivities. Overall, thermal processing caused a significant reduction of the health-promotion effects of beans.     

Phenolic composition and mammary cancer cell inhibition of extracts of whole cowpeas (Vigna unguiculata) and its anatomical parts

The phenolic profile and anticancer properties of extracts from whole cowpeas, seed coats and cotyledons were determined. Seed coats contained at least 5 and 10 times more free and bound phenolics compared to whole seeds (75.6 and 31.7 mg/100 g of free and bound phenolics, respectively). Seed coats and cotyledons contained about 50% and 95% of free phenolics, respectively. The major phenolics associated to seed coats were gallic and protocatechuic acid, whereas in cotyledons p-hydroxybenzoic acid was prevalent. Seed coats contained approximately 10 times more flavonoids compared to whole seeds and cotyledons were practically free of flavonoids. After acid hydrolysis, myricetin, quercetin and kaempferol were identified in seed coats. Most of the antioxidant activity determined with the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) assays was exerted by free phenolics. The free phenolic extract of whole seeds at a concentration of approximately 100 mg gallic acid equiv. (GAE)/l inhibited 65% the proliferation of hormone-dependent mammary (MCF-7) cancer cells. Extracts of seed coats or cotyledons also inhibited cell proliferation but to a significantly lesser extent, thus indicating synergistic effects between phenolics and other phytochemicals associated to these anatomical parts.     

In vitro scavenging capacity of annatto seed extracts against reactive oxygen and nitrogen species

Bixa orellana L. (annatto), from Bixaceae family, is a native plant of tropical America, which accumulates several carotenoids (including bixin and norbixin), terpenoids, tocotrienols and flavonoids with potential antioxidant activity. In the present study, the in vitro scavenging capacity of annatto seed extracts against reactive oxygen species (ROS) and reactive nitrogen species (RNS) was evaluated and compared to the bixin standard. Annatto extracts were obtained using solvents with different polarities and their phenolic compounds and bixin levels were determined by high performance liquid chromatography coupled to diode array detector. All annatto extracts were able to scavenge all the reactive species tested at the low μg/mL range, with the exception of superoxide radical. The ethanol:ethyl acetate and ethyl acetate extracts of annatto seeds, which presented the highest levels of hypolaetin and bixin, respectively, were the extracts with the highest antioxidant capacity, although bixin standard presented the lowest IC₅₀ values.     

Fatty acid composition,oxidative stability,antioxidant and antiproliferative properties of selected cold-pressed grape seed oils and flours

Cold-pressed chardonnay, muscadine, ruby red, and concord grape seed oils and their defatted flours were studied for their fatty acid composition, oxidative stability and antioxidant and antiproliferative activities. The phenolic profiles of the seed flours were also measured. The most abundant fatty acid in the oils was linoleic acid, ranging from 66.0 g/100 g of total fatty acids in ruby red seed oil to 75.3 g/100 g of total fatty acids in concord seed oil. The oils were also high in oleic acid and low in saturated fat. Ruby red grape seed oil recorded the highest oxidative stability index of 40 h under the accelerated conditions. Total phenolic content (TPC) was up to 100 times lower in the oils than in the flours. Lutein, zeaxanthin, cryptoxanthin, β-carotene, and α-tocopherol levels were also measured. DPPH radical-scavenging capacity ranged from 0.07 to 2.22 mmol trolox equivalents (TE)/g of oil and 11.8 to 15.0 mmol TE/g of flour. Oxidative stability of menhaden fish oil containing extracts of the seed flours was extended by up to 137%. HPLC analysis was conducted to determine the levels of free soluble, soluble conjugated and insoluble bound phenolics in the seed flours. The phenolic compounds analyzed included catechin, epicatechin, epicatechin gallate, quercetin, gallic acid, and procyanidins B1 and B2. Antiproliferative activity was tested against HT-29 colon cancer cells. All of the seed flours and muscadine seed oil registered significant (P < 0.05) inhibition of cancer cell growth. The results from this study demonstrate the potential of developing value-added uses for these seed oils and flours as dietary sources of natural antioxidants and antiproliferative agents for optimal health.     

青钱柳叶活性成分的抗氧化活性及UPLC-QTOF-MS/MS分析

利用96孔板法测定青钱柳叶不同溶剂(水、70%乙醇、乙酸乙酯和正己烷)提取物中总酚、总黄酮含量及抗氧化活性(DPPH自由基清除能力、还原能力、总抗氧化能力),考察酚类物质含量与抗氧化活性的相关性,并采用超高效液相色谱-四极杆飞行时间串联质谱(ultra performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spetrometry,UPLC-QTOF-MS/MS)分析提取物中主要活性成分。结果表明:不同提取物中总酚、总黄酮含量及抗氧化活性存在显著性差异且70%乙醇溶液提取物表现出最高的总酚(219.01 mg GAE/g)、总黄酮含量(7.23 mg CE/g)及最强的DPPH自由基清除能力(35.46 mg TE/g)和还原能力(1.89 mmol Fe SO_4/g);总酚、总黄酮含量与DPPH自由基清除能力、还原能力之间呈正相关,与总抗氧化能力呈显著负相关,表明多酚类物质是青钱柳中主要的抗氧化剂。UPLC-QTOF-MS/MS分析70%乙醇溶液提取物并初步鉴定出22种化合物,包括2种有机酸、4种酚酸、5种黄酮、8种三萜皂苷类和3种酯类,其中酚酸和黄酮类化合物是主要的抗氧化活性成分,有机酸、三萜皂苷及酯类化合物可能是潜在的抗氧化活性成分。     

The effect of sorghum type and processing on the antioxidant properties of African sorghum-based foods

This work determined the effect of sorghum type and different processing technologies of traditional African sorghum foods on total phenols, tannin content and antioxidant activity. The products were prepared by fermentation, conventional and extrusion cooking of whole and decorticated ground grain. The tannin sorghum types, had higher ABTS and DPPH antioxidant activities, compared to the types without tannins. Antioxidant activity was significantly correlated with total phenols and tannins (r > 0.95). Decortication, reduced antioxidant activity of both tannin and non-tannin sorghum by 82–83% due to the removal of the pericarp and the testa, which decreased phenols. Processing, generally decreased antioxidant activity, however, conventionally cooked porridges had higher antioxidant activity than the extrusion cooked products. The retention of antioxidant activity, particularly in fermented and unfermented porridges, means that whole tannin sorghum can be processed into foods with potential health benefits.     

Determination of Polyphenols,Flavonoids, and Antioxidant Capacity in Colored Chickpea (Cicer arietinum L.)

ABSTRACT: Dry legumes are staple and potentially functional food, being a good source of polyphenols, flavonoids, and antioxidant activity. The objective of this study was to determine the total polyphenol content (TPC), total flavonoid content (TFC), and their relation with antioxidant capacity in 17 chickpea lines having colored seed coats (black, red, brown, green, rubiginous, gray, yellow, cream, or beige). The seed coat usually contains more than 95% of these compounds. In this study, both TPC and TFC varied significantly among different lines and were highly correlated to antioxidant activity. Colored seeds contained up to 13-, 11-, and 31-fold more TPC, TFC, and antioxidant activity, respectively, than cream- and beige-color seeds. Thus, colored chickpea could be a potentially functional food in addition to its traditional role of providing dietary proteins and dietary fibers.     

Antioxidant and antiproliferative activities of mango (Mangifera indica L.) flesh and peel

The antioxidant and antiproliferative properties of flesh and peel of mango (Mangifera indica L.) were investigated. The cytoprotective effect of mango flesh and peel extracts on oxidative damage induced by H₂O₂ in a human hepatoma cell line, HepG2, were determined, and the underlying mechanism was examined by a single-cell electrophoresis assay (comet assay). Treatment of HepG2 cell with mango peel extract prior to oxidative stress was found to inhibit DNA damage. The free radical scavenging activities of mango flesh and peel extracts were evaluated by electron spin resonance (ESR). The mango peel extract exhibited stronger free radical scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and alkyl radicals than mango flesh extract, regardless of ripeness. Similarly, peel extract exhibited significant antiproliferative effect against all tested cancer cell lines, compared to that of flesh extract, in a dose-dependent manner. The result also showed that the antiproliferative activity of mango flesh and peel extracts correlated with their phenolic and flavonoid contents. Thus, mango peel, a major by-product obtained during the processing of mango product, exhibited good antioxidant activity and may serve as a potential source of phenolics with anticancer activity.     

Comparative study on antiproliferation properties and cellular antioxidant activities of commonly consumed food legumes against nine human cancer cell lines

The aims of this work were to compare health promoting effects of commonly consumed food legumes in terms of cancer cell proliferation inhibitory effects and cellular antioxidant activities (CAA). The CAA was evaluated by fluorescence microplate reader based on in vitro animal cell cultivation. Antiproliferative properties were assayed by MTT method using in vitro cell culture system. Phytochemicals (including total phenolic, procyanidin, saponin and phytic acid) and chemical antioxidant activities (including DPPH free radical scavenging activity, oxygen radical absorbing capacity, peroxyl radical scavenging capacity (PRSC)) were also determined for comparison purposes. The results showed that different types of legumes possessed considerable variations in their phytochemicals, as well as chemical and cellular antioxidant activities. Adzuki bean exhibited the strongest antiproliferative properties in a dose-dependent manner against all digestive system cancer cell lines (CAL27, AGS, HepG2, SW480 and Caco-2), ovary cancer cell SK-OV-3 and breast cancer cell MCF-7 among all legumes tested. Black soybean exhibited the highest saponin, phytic acid content, PRSC values, and the strongest CAA values. These results indicate that commonly consumed food legumes may serve as an excellent dietary source of natural antioxidants for health promotion and cancer prevention.     

Rind of the rambutan, Nephelium lappaceum, a potential source of natural antioxidants

The rind of rambutan, which is normally discarded was found to contain extremely high antioxidant activity when assessed using several methods. Although having a yield of only 18%, the ethanolic rambutan rind extract had a total phenolic content of 762 ± 10 mg GAE/g extract, which is comparable to that of a commercial preparation of grape seed extract. Comparing the extract’s pro-oxidant capabilities with vitamin C, α-tocopherol, grape seed and green tea, the rind had the lowest pro-oxidant capacity. In addition, the extract at 100 μg/ml was seen to limit oxidant-induced cell death (DPPH at 50 μM) by apoptosis to an extent similar to that of grape seed. The extracts were not cytotoxic to normal mouse fibroblast cells or splenocytes while the powderised rind was seen to have heavy metals contents far below the permissible levels for nutraceuticals. Our study for the first time reveals the high phenolic content, low pro-oxidant capacity and strong antioxidant activity of the extract from rind of Nephelium lappaceum. This extract, either alone or in combination with other active principles, can be used in cosmetic, nutraceutical and pharmaceutical applications.     

Identification of bioactive response in traditional cherries from Portugal

In recent years many studies on cherries have revealed that they are rich sources of bioactive compounds with potential human health benefits. In this work, we evaluated the antioxidant activity and antiproliferative effect in human cancer cells of nine sweet cherries, including two traditional cultivars from Portugal (Saco and Morangão). Results obtained in biological assays, together with the phenolic composition of cherries, were submitted to principal component analysis (PCA) which allowed samples to be grouped in terms of their bioactivity. Saco cherry and two exotic cultivars (Ulster and Lapin) proved to have higher contents of phenolic compounds, highest antioxidant activity and were the most effective in inhibiting human cancer cells derived from colon (HT29) and stomach (MKN45). Correlation of the data obtained showed that anthocyanins were the major contributors to the antioxidant capacity and antiproliferative effect of cherries. Additionally, hydroxycinnamic acids (neochlorogenic acid, chlorogenic acid and p-coumaroylquinic acid), flavan-3-ols (catechin and epicatechin) and flavonols (rutin and quercetin-3-glucoside) also play important roles in protection against oxidative stress.