Quantification of the polyphenolic fraction and in vitro antioxidant and in vivo anti-hyperlipemic activities of Hibiscus sabdariffa aqueous extract

In the present study the quantification of the polyphenolic fraction, anthocyanins and other polar compounds, the antioxidant capacity and the anti-hyperlipemic action of the aqueous extract of Hibiscus sabdariffa has been achieved. Seventeen compounds were successfully quantified either by HPLC-DAD or HPLC-ESI-TOF-MS. Six of them were directly quantified by their corresponding standards, whereas the rest were indirectly quantified as equivalents using standards of similar compounds. The antioxidant capacity have also been estimated by comparing different assays, i,e, Trolox equivalent antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and measurement of thiobarbituric acid reacting substances (TBARS). H. sabdariffa showed high reducing capacity in FRAP assay and significant capability to scavenge peroxyl radicals in the ORAC assay. Nevertheless, the extract exhibited poor efficacy to inhibit peroxyl radicals in lipid systems. The plant extract also exhibited the capacity to decrease serum triglyceride concentration on hyperlipemic mouse model.     

Antioxidant activities of malt extract from barley (Hordeum vulgare L.) toward various oxidative stress in vitro and in vivo

The antioxidant activities of malt extract from barley were evaluated by various methods in vitro and in vivo. Scavenging effects on the hydroxyl and superoxide radicals, and protection against reactive oxygen species induced lipid, protein and DNA damage were evaluated. The d-galactose induced mouse aging model was used to evaluate ability of malt extract to behave as an antioxidant in vivo. The extract exhibited high antioxidant activities both in vitro and in vivo, evidenced by its ability to scavenge hydroxyl- and superoxide-radicals, high reducing power, and protection against biological macromolecular oxidative damage. Furthermore, malt extract prevented the decrease of antioxidant enzyme activities, decreased liver and brain malondialodehyde levels and carbonyl content, and improved total antioxidant capability in d-galactose-treated mice. In conclusion, these results demonstrate potential antioxidant activities and antiaging effect of malt, providing scientific support for the empirical use of malt as an antioxidant for diseases caused by reactive oxygen species.     

A comparative study of tocopherols composition and antioxidant properties of in vivo and in vitro ectomycorrhizal fungi

In aerobic organisms, the free radicals are constantly being produced during the normal cellular metabolism. The antioxidant properties of many organisms and particularly of wild mushrooms with their content in antioxidant compounds such as tocopherols, can detoxify potentially damaging forms of activated oxygen. Herein, a comparative study of tocopherols composition and antioxidant properties of in vivo (fruiting bodies) and in vitro (mycelia) ectomycorrhizal fungi: Paxillus involutus and Pisolithus arhizus. Tocopherols were determined by high performance liquid chromatography (HPLC) coupled to a fluorescence detector. The antioxidant properties were studied in terms of DPPH radical-scavenging activity, reducing power and inhibition of β-carotene bleaching. Fruiting bodies revealed the highest antioxidant properties, including scavenging effects on free radicals (EC₅₀ = 0.61 and 0.56 mg/ml) and inhibition of lipid peroxidation capacity (EC₅₀ = 0.40 and 0.24 mg/ml for P. involutus and P. arhizus, respectively), than mycelia produced in vitro cultures. Nevertheless, mycelia revealed higher levels of total tocopherols than fruiting bodies, and particularly P. arhizus mycelium proved to be a powerful source of γ-tocopherol (154.39 μg/g dry weight).     

In vivo and in vitro antidiabetic effects of aqueous cinnamon extract and cinnamon polyphenol-enhanced food matrix

Cinnamon has a long history of medicinal use and continues to be valued for its therapeutic potential for improving metabolic disorders such as type 2 diabetes. In this study, a phytochemically-enhanced functional food ingredient that captures water soluble polyphenols from aqueous cinnamon extract (CE) onto a protein rich matrix was developed. CE and cinnamon polyphenol-enriched defatted soy flour (CDSF) were effective in acutely lowering fasting blood glucose levels in diet induced obese hyperglycemic mice at 300 and 600 mg/kg, respectively. To determine mechanisms of action, rat hepatoma cells were treated with CE and eluates of CDSF at a range of 1–25 μg/ml. CE and eluates of CDSF demonstrated dose-dependent inhibition of hepatic glucose production with significant levels of inhibition at 25 μg/ml. Furthermore, CE decreased the gene expression of two major regulators of hepatic gluconeogenesis, phosphoenolpyruvate carboxykinase and glucose-6-phosphatase. The hypoglycemic and insulin-like effects of CE and CDSF may help to ameliorate type 2 diabetes conditions.     

Soymilk phenolic compounds,isoflavones and antioxidant activity as affected by in vitro gastrointestinal digestion

The aim of this research was to evaluate changes in the phenolic compounds, isoflavones and antioxidant activity of soymilk following in vitro gastrointestinal digestion (including dialysis). Gastric digestion significantly influenced the release of bioactive substances from the soymilk matrix, increasing the concentration of total phenolic components (35% as the sum of individuals and 14% by Folin–Ciocalteu [F–C] method), total isoflavone content (22%) and total antioxidant activity (76%). The concentration of all those compounds was reduced significantly in the duodenal fraction in comparison to gastric digestion and their lowest concentration was observed in the dialysed fraction, where phenolic acids were not detected. The bioaccessibility of soymilk phenolic compounds was 15% as the sum of individuals and 20% by F–C assay; isoflavones 36% and constituents with antioxidant activity 27%. Results suggest that most of these compounds were sufficiently available to be absorbed and could contribute health benefits.     

The in vitro and in vivo antioxidant properties of seabuckthorn (Hippophae rhamnoides L.) seed oil

The antioxidant capacity of seabuckthorn (Hippophae rhamnoides L.) seed oil was investigated with a number of established in vitro assays and in an in vivo study of carbon tetrachloride (CCl₄)-induced oxidative stress in mice. The results showed that DPPH radical scavenging activity, ferrous ion chelating activity, reducing power and inhibition of lipid peroxidation activity all increased with increasing concentrations of seabuckthorn seed oil. Moreover, the EC₅₀ values of seabuckthorn seed oil from the hydrogen peroxide, superoxide radical, hydroxyl radical scavenging assays were 2.63, 2.16 and 0.77 mg/ml, respectively. In the in vivo study, seabuckthorn seed oil inhibited the toxicity of CCl₄, as seen from the significantly increased activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The GSH content in the liver was also increased, whereas hepatic malondialdehyde was reduced. Taken together, these results clearly indicate that seabuckthorn seed oil has significant potential as a natural antioxidant agent.     

In vitro antioxidant activity of liquor and semi-purified fractions from fermented squid pen biowaste by Serratia ureilytica TKU013

Antioxidative activities were found in the culture supernatant of Serratia ureilytica TKU013 with squid pen as the sole carbon/nitrogen source. The 4th day supernatant showed the strongest antioxidant activities and the highest total phenolic content. The supernatant was further purified by liquid–liquid partition, and it was found that the ethyl acetate-soluble (EA) extract exhibited the strongest antioxidant activity in a DPPH radical-scavenging assay and this was compared with the positive control, α-tocopherol. This extract was further divided into eight fractions, designated as F1–F8, by silica gel liquid chromatography. In most cases, F4 and F8 were found to possess the strongest antioxidative activities. Significant associations between the antioxidant potency and the total phenolic content, as well as between the antioxidant potency and free amino groups, were found for the supernatant, extract, and fractions. With this method, squid pen waste can be utilized and has potential in the production of functional foods.     

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.     

Chemical composition, mineral content and antioxidant activity of Verbena officinalis L.

Aqueous and hydroalcoholic extracts from Verbena officinalis L. were obtained and characterised. The analysis by HPLC-DAD and LC-MS allowed the detection and identification of three iridoids, fifteen flavonoids and four phenolic acid derivatives. Four flavonoids, scutellarein 7-diglucuronide (9), scutellarein 7-glucuronide (13), pedalitin 6-galactoside (15) and scutellarein 7-glucoside (19) are reported for the first time from this plant. In addition, three new flavonoids have been isolated: scutellarein 7-O-(2-O-feruloyl)-diglucuronide (5), pedalitin 6-O-diglucuronide (6) and pedalitin 6-O-(2-O-feruloyl)-diglucuronide (13). To our knowledge, these flavonoids have not been reported as natural products. Both extracts showed significant antioxidant activity using three in vitro model systems and the results have been correlated with total phenolic and total flavonoid contents. The results have allowed establishing an important relation structure-activity and significant correlations have also been found between the mineral content and the flavonoids present in both extracts.     

Polyphenolic content and antioxidant activity of Eugenia pollicina leaf extract in vitro and in model emulsion systems

The polyphenolic profile of a leaf extract of the Mauritian endemic plant, Eugenia pollicina, was assessed as a source of natural antioxidants. The amounts of flavan-3-ol derivatives determined by HPLC, were in the order of (−)-epicatechin (EC) > (−)-epigallocatechin gallate (EGCG) > (+)-catechin (C) > (−)-epicatechin gallate (ECG) with the levels of Procyanidin B2 and B1 dimers ranging from 1 to 3 mg g⁻¹ FW. The trolox equivalent antioxidant capacity and ferric reducing antioxidant power values were 796 ??mol g⁻¹ FW and 302 ??mol g⁻¹ FW respectively. E. pollicina extracts also strongly inhibited the FeCl₃ and ascorbate-dependent microsome lipid peroxidation, a function that is linked to their flavonoid contents. The extent of DNA damage induced by the extract under study in the copper-phenanthroline assay was lower than the effect of a reference of 240 ??M ascorbate. E. pollicina extracts also inhibited lipid autoxidation in the 30% (v/v) olive oil and soybean oil oil-in-water (O/W) emulsions and was effective in slowing down the formation of hydroperoxides in the emulsions during 13 days storage at 40 °C as determined by the peroxide, conjugated diene and para-anisidine values. The high levels of total phenolics, flavonoids and procyanidins measured indicate that E. pollicina is a potential source of antioxidants relevant to the maintenance of oxidative stability of the food matrix, cosmetics and/or pharmaceutical preparations.     

Phenolics from hull of Garcinia mangostana fruit and their antioxidant activities

The air-dried fruit hulls of Garcinia mangostana Linn. were extracted with 70% MeOH, and then partitioned into the n-BuOH fractions. Furthermore, three major phenolic components related to their antioxidant activities were purified by silica gel column chromatography and Sephadex LH-20 and then identified as P₁ (1,3,6,7-tetrahydroxy-2,8-(3-methyl-2-butenyl)), P₂ [1,3,6-trihydroxy-7-methoxy-2,8-(3-methyl-2-butenyl) xanthone] and P₃ (epicatechin) using UV–visible spectrophotometry, IR spectrophotometry and NMR spectroscopy, respectively. The antioxidant activities of three major phenolics were evaluated using different tests, including the free radical scavenging capability and total antioxidant activity in a linoleic acid peroxidation. These three phenolic compounds exhibited different antioxidant activities in these antioxidant tests. The hydroxyl radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capabilities and the activity against linoleic acid peroxidation of P₁ were greater than those of P₂ and P₃, while the superoxide anion radical scavenging activity of P₃ was greater than that of P₁, but was close to that of P₂ or α-tocopherol. An activity-guided isolation and purification process was used to identify the components showing the strong DPPH radical scavenging activity from G. mangostana Linn.     

Profiling of in vitro neurobiological effects and phenolic acids of selected endemic Salvia species

The ethyl acetate and methanol extracts from 16 Salvia L. species were screened for their inhibitory activity against acetylcholinesterase, butyrylcholinesterase, lipoxygenase, and tyrosinase; the enzymes linked to neurodegeneration. Their antioxidant activity was also tested using DPPH radical scavenging, metal-chelation, and ferric-reducing antioxidant power (FRAP) assays. Total flavonoid content of the extracts was determined by AlCl₃ reagent, while HPLC technique was applied for analysis of various phenolic acids in the extracts. The extracts exerted weak cholinesterase and tyrosinase inhibition, and remarkable inhibition against lipoxygenase (13.07 ± 2.73-74.21 ± 5.61%) at 100 μg ml⁻¹. The methanol extracts showed higher antioxidant activity in DPPH radical scavenging and FRAP assays. The extracts were analyzed for their gallic, protocateuchic, p-hydroxy-benzoic, vanillic, caffeic, chlorogenic, syringic, o- and p-coumaric, ferulic, rosmarinic, and tr-cinnamic acid contents and the methanol extract of Salvia ekimiana (153.50 mg 100 g⁻¹) was revealed to be the richest in terms of rosmarinic acid.     

In vitro antioxidative activities of extract and semi-purified fractions of the marine red alga, Rhodomela confervoides (Rhodomelaceae)

The methanol–chloroform extract of the marine red alga, Rhodomela confervoides, was measured for antioxidant activity, using the α,α-diphenyl-β-picrylhydrazyl radical-scavenging assay and the β-carotene-linoleate bleaching assay systems, and compared with those of the positive controls of butylated hydroxytoluene, gallic acid and ascorbic acid. The active extract was further purified by liquid–liquid partition to afford four fractions, of which the ethyl acetate-soluble (EA) fraction exhibited the strongest antioxidant activity in both assay systems. This fraction was further divided into seven subfractions, designated as EA1–EA7, by silica gel vacuum liquid chromatography. In most cases, EA1 and EA4 were found to possess the strongest activity. The total phenolic contents and reducing powers of the extract, fractions, and subfractions were also determined. Significant associations between the antioxidant potency and the total phenolic content, as well as between the antioxidant potency and the reducing power, were found for the tested fractions and subfractions.     

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.     

Antioxidant activity of stilbene glycoside from Polygonum multiflorum Thunb in vivo

Stilbene glycosides were isolated from the ethanol extract of the roots of Polygonum multiflorum Thunb. Two samples were obtained; a fraction separated by macroporous resin and pure crystals of 2,3,5,4′-tetrahydroxystilbene 2-O-β-glucopyranoside. The antioxidant activities of these two samples were evaluated using antioxidant tests of rats in vivo. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of the serum and the organs (liver, heart and brain of rats) of d-galactose induced senile rats which were fed with stilbene glycoside, were increased; however, the content of 2-thiobarbituric acid-reactive substances (TBARS) was decreased. It is concluded that the stilbene glycoside from Polygonum multiflorum Thunb possesses high in vivo antioxidant activity.     

Evaluation of the antioxidant activity of five endemic Ligustrum species leaves from Taiwan flora in vitro

Leaves from the plant species belonging to the genus Ligustrum are widely used as tea or herbal medicine in Europe, China, and Japan. The antioxidant properties of five Ligustrum species from Taiwan were compared using in vitro antioxidant methods such as DPPH radical scavenging, TEAC, and FRAP assays. Cell-based antioxidant methods were used, including Fe²⁺/ascorbate-induced lipid peroxidation on brain homogenate and AAPH-induced erythrocyte haemolysis. The amounts of major phenolic compounds from the Ligustrum species, including phenylpropanoids, flavonoids, and iridoids, were determined by spectrophotometric methods. The results showed that all Ligustrum species exhibited antioxidant, radical-scavenging, anti-haemolytic, and lipid peroxidation-inhibiting activities at different magnitudes of potency. A significant correlation was found between antioxidant activity and the amount of antioxidant components, in particular, total phenolics and phenylpropanoids. Among all Ligustrum species from Taiwan, Ligustrum morrisonense is presented as potential source of natural antioxidants.