Characterisation and free radical scavenging activities of novel red pigment from Osmanthus fragrans’ seeds

A novel red pigment (RP) was isolated from Osmanthus fragrans  ’ seeds. The optimised experimental parameters of extraction obtained with a four-factor at three-level orthogonal array experimental design _L9(3⁴)$L_9(3^4)$ were ethanol concentration, temperature, pH and extraction time as 90%, 78 °C, 2.5 and 40 min, respectively. A yield of 34.6 ± 2.2 g/100 g was obtained under optimised conditions. The red pigment directly from O. fragrans’ seeds can be dissolved in alkaline, acidic waters solutions and hydrophilic organic solvents in common use. The colour of a water solution of RP changed with pH. RP was stable to heat in the temperature range of 25–100 °C. Physical and chemical properties of RP revealed that the red pigment was also stable in the presence of Na₂SO₃, NaCl, amino acid, organic acid, sugar, starch or metal-ions (such as Ca²⁺, Cu²⁺, Fe³⁺, Zn²⁺, Al³⁺, Mg²⁺ and Na⁺), but was bleached by strong oxidants (KMnO₄, K₂Cr₂O₇ and NaOCl). Subsequently, free radical scavenging activities of RP were assessed using 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals using a new resonance scattering spectral method. RP showed an excellent DPPH radical scavenging activity and was superior to butylated hydroxytoluene (BHT), and exhibited quite a strong concentration-dependent inhibition of hydroxyl radical at low concentrations compared with ascorbic acid and quercetin. When the concentration of RP was 0.03 μg/ml, the scavenging percentage of hydroxyl radical reached 92.3%. Salidroside was isolated as an active principle.     

Antioxidant capacity of extract from edible flowers of Prunus mume in China and its active components

Prunus mume flowers are used as traditional edible and medicinal materials in China. In this study, phenolic compounds and antioxidant activity of ethanolic extract from flowers of P. mume in China were investigated for the first time. The total phenol content was estimated as gallic acid equivalents by the Folin-Ciocalteu reagent method. The antioxidant activity was measured by DPPH, ABTS⁺, and OH free radicals scavenging and ferric-reducing antioxidant power (FRAP). Three chlorogenic acid isomers, namely, 3-O-caffeoylquinic, 4-O-caffeoylquinic and 5-O-caffeoylquinic acids, were isolated and purified by preparative HPLC from the ethanolic extract and identified by UV, MS and NMR. The contents of these isolated compounds were quantified by HPLC. Results showed that 5-O-caffeoylquinic acid was of the highest level in these three isomers. The ethanolic extract demonstrated activity to some degree in all the antioxidant assays. In all tested assays, all of the isolated chlorogenic acid isomers exhibited strong antioxidant activities, which were almost the same. The results showed that chlorogenic acid isomers are the key phenolic compounds which are responsible for antioxidant activity of the ethanolic extract from Chinese P. mume flowers.     

Antioxidant and free radical scavenging activities of phenols from onion (Allium cepa)

Four (red, violet, white and green) varieties of Allium cepa were studied for their total phenolic contents (TPC), antioxidant (AOA) and free radical scavenging activities (FRSA). The TPC varied from 4.6 to 74.1 mg/g GAE, AOA varied from 13.6% to 84.1% and FRSA showed wide range in terms of IC₅₀ (inhibitory concentration) from 0.1 to 15.2 mg/ml, EC₅₀ (efficient concentration) from 4.3 to 660.8 mg/mg and ARP (antiradical power) from 0.15 to 23.2. The outer dry layers of red and violet varieties showed better inhibition of lipid peroxidation assayed by ammonium thiocyanate than α-tocopherol. The non-site-specific inhibition of hydroxyl radical induced deoxyribose degradation was also higher in the outer dry layers of red and violet varieties than in their middle and inner layers. The outer layers were also potential inhibitors of nitroblue tetrazolium chloride (NBT) reduction caused by superoxide anions. On the other hand the ferrous ion chelating capacity of the red and violet varieties was highest in the inner layers. Specific phenolic composition performed through HPLC and LC–MS/MS showed the presence of gallic acid, ferulic acid, protocatechuic acid, quercetin, and kaempferol. The unutilised outer layers of the red variety were a rich source of quercetin (5110 μg/g) with high AOA, FRSA and also showed significant protection of DNA damage caused by free radicals.     

Variations in the components of Osmanthus fragrans Lour. essential oil at different stages of flowering

A headspace solid-phase microextraction (HS-SPME) method was used to extract the essential oil of Osmanthus fragrans Lour., which was then analysed by gas chromatography–mass spectrometry (GC–MS) at four different stages of flowering. The primary chemical components of the essential oil extracted from O. fragrans were linalool and its oxide, α-ionone, β-ionone, nerol, γ-decalactone, 9,12,15-octadecatrienoic acid, and hexadecanoic acid, most of which were at their highest concentrations in extracts obtained from flowers at the initial flowering stage. HS-SPME using a PDMS/DVB fibre is a simple, rapid, and solvent-free method for the extraction of volatile compounds emitted from living O. fragrans. HS-SPME with GC–MS can be used to determine the aromatic maturity in O.fragrans, to decide the optimal harvest date. The initial flowering stage was the best time to harvest O. fragrans.     

Antioxidant activity and melanogenesis inhibitory effect of the acetonic extract of Osmanthus fragrans: A potential natural and functional food flavor additive

Osmanthus fragrans, a common flavor additive for tea and other beverages, has potential applications in biomedical science. In this study, we investigated O. fragrans acetonic extract (OFE) for its total phenolic and flavonoid contents, radical-scavenging activity, and potential anti-tyrosinase ability. OFE possessed considerable amounts of phenolics (264.7 mg gallic acid equivalent/g of extract) and flavonoids (190.7 mg catechin equivalent/g of extract). The antioxidation activities, measured in terms of EC₅₀ values using DPPH and ABTS⁺ assays, were 304.9 mg ascorbic acid equivalent/g of extract and 516.3 mg trolox equivalent/g of extract, respectively. LC-MS results discovered the presence of luteolin in the extract, and a kinetic study revealed an uncompetitive inhibitory effect of OFE upon the oxidation of tyrosine (IC₅₀ = 2.314 mg/mL) and l-DOPA (IC₅₀ = 44.20 mg/mL). In addition, we tested OFE in vitro (B16F10 cells) for its anti-tyrosinase activity and anti-melanin formation and found that OFE was able to reduce the tyrosinase activity and melanin formation of B16F10 cells in a dose-dependent manner. Our findings support that O. fragrans is a potential natural, functional antioxidant food favor additive. Additionally, because OFE inhibits melanin formation, it appears to have potential use as an anti-browning food additive, in skin-whitening cosmetics, or as a new drug for treating melanoma.     

Antioxidant activities of Salvia miltiorrhiza and Panax notoginseng

Traditional Chinese medicinal herb Salvia miltiorrhiza (SM) and Panax notoginseng (PN) have been widely used for the prevention and treatment of vascular diseases in the clinics. To better understand their mechanisms of pharmacological actions, the in vitro antioxidant activities of extract of Salvia miltiorrhiza (ESM) and extract of Panax notoginseng (EPN) were evaluated with different antioxidant testing systems. Their activities of scavenging superoxide anion radicals, DPPH radicals, hydroxyl radicals, and hydrogen peroxide, chelating Ferrous ion, and ferric ion reducing power were assessed. The results showed that the mechanisms of their antioxidant effects were distinct and diverse. ESM possessed strong reducing power and high scavenging activities against free radicals including superoxide anion, hydroxyl and DPPH radicals, but a weaker scavenging activity for hydrogen peroxide. ferrous ion chelating activity of ESM was undetectable at the tested concentrations. In contrary, EPN exhibited strong ferrous ion chelating activity and high scavenging activities against hydrogen peroxide, hydroxyl radicals, and a weak activity against superoxide anion and DPPH free radicals. EPN did not show any ferric ion reducing power. Since their antioxidant mechanisms are complementary, the combined use of ESM and EPN might be even more beneficial. These antioxidant properties of SM and PN are likely part of the reasons that they are effective in the prevention and treatment of vascular diseases.     

Antioxidant and free radical scavenging activities of polyphenol-enriched curry leaf (Murraya koenigii L.) extracts

The in vitro antioxidant properties of different extracts (water, alcohol, alcohol:water, hexane or chloroform extract) of curry leaves (Murraya koenigii L.) were evaluated using various assays. The alcohol:water (1:1) extract of curry leaves (AWEC) showed the highest antioxidant and free radical scavenging activity. It inhibited membrane lipid peroxidation by 76%, at 50 μg/ml, scavenged 93% of superoxides at 200 μg/3 ml and scavenged approximately 90% of hydroxyl and 1,1-diphenyl-2-picrylhydrazayl radicals at 4–5-fold lower concentrations compared to the other tested extracts. In addition, the alcohol:water extract reduced cytochrome c and ferric ion levels, chelated ferrous ions and inhibited ferrous sulfate:ascorbate-induced fragmentation and sugar oxidation of DNA. These results establish the antioxidant potential of AWEC, which could be used as natural antioxidant source.     

Antioxidant activity of the roots of Decalepis hamiltonii (Wight & Arn.)

Tuberous roots of Decalepis hamiltonii (Dh) are consumed in southern India as pickles and beverage for their health benefits. The antioxidant potential of the roots was measured using various in vitro assays. Among the extracts, the methanolic and aqueous extracts showed high antioxidant activity measured as scavenging of DPPH, superoxide and hydroxyl radicals. Both the aqueous and methanolic extracts inhibited microsomal lipid peroxidation and exhibited strong reducing power and metal chelating activity. The antioxidant activity did not correlate with the phenolic content of the extracts. These results demonstrate the antioxidant potency of the root extracts which could be the basis for its alleged health promoting potential of Dh. The roots of Dh could serve as a new source of natural antioxidants or nutraceuticals with potential applications to reducing the level of oxidative stress and related health benefits.     

Evaluation of reducing power and radical scavenging activities of water and ethanol extracts from sumac (Rhus coriaria L.)

The purpose of this study was to determine the reducing power, metal chelating, and radical scavenging capabilities of water and ethanol extracts of sumac (Rhus coriaria L.), comparatively. The water and ethanol extracts of sumac were evaluated for their radical scavenging activities by means of the DPPH and DMPD assays. Water extract of sumac (R. coriaria L.) scavenged radicals effectively with EC₅₀ values of 36.4 ??g/ml for DPPH free radical and 44.7 ??g/ml for DMPD cation radical. Similarly, the total reducing power of water extract was found higher than ethanol extract in both potassium ferricyanide reduction (FRAP) and cupric ions reduction capacity methods (CUPRAC). 2,2′-Bipyridine was used to determine the metal chelating activity and the result of water extract was found higher than ethanol extract. Total phenolic and total flavonoid contents of both extracts were studied as well. The values of water extract were found to be higher than that of ethanol extract. The present study found that water extracts of sumac (R. coriaria L.) have effective antioxidant and radical scavenging activities as compared to ethanol extracts.     

Isolation and characterization of melanin from Osmanthus fragrans’ seeds

Melanin derived from Osmanthus fragrans’ seeds was isolated from O. fragrans’ seeds by alkaline extraction, acid hydrolysis, and repeated precipitation with a yield of 0.34 g/100 g (wet weight basis). Physical and chemical properties of the melanin revealed that the melanin derived directly from O. fragrans’ seeds were similar to typical melanin. The melanin was stable under ultraviolet light or room-light, stable in the range of 25-100 °C, relatively stable in alkaline solution, reducer and salt, but was bleached by strong oxidants (KMnO₄, K₂Cr₂O₇ and NaOCl). The metal-ion of Ca²⁺, Cu²⁺, Fe³⁺ and Zn²⁺ had the function of color increase or color preservation to the melanin. Although Mg²⁺, Al³⁺ and Na⁺ reduce pigment color, it was not obvious. Amino acid and organic acid did not affect the melanin, while sugar, starch, and glucose affected it slightly.     

Free radical scavenging activity and phenolic content in achenes and thalamus from Fragaria chiloensis ssp. chiloensis, F. vesca and F. x ananassa cv. Chandler

The total phenolic, flavonoid and anthocyanin content of achenes (true fruit) and thalamus (receptacle) from the native South American Fragaria chiloensis ssp. chiloensis (f. patagonica and f. chiloensis), Fragaria vesca and Fragaria x ananassa cv. Chandler was determined by spectrophotometric means. Highest phenolic content was found in F. vesca while lowest content was measured for white strawberry (F. chiloensis ssp. chiloensis, f. chiloensis). The total anthocyanin and total flavonoid contents in the samples investigated was lower for the white strawberry and higher in F. x ananassa cv. Chandler. Total flavonoid content showed a better correlation than total anthocyanins with the free radical scavenging effect of the extracts measured by means of the DPPH discoloration assay. In the superoxide anion assay all the acetone extracts of strawberries showed similar activity. The data presented in this study demonstrate that the amount of phenolic compounds differ significantly between species and subspecies and determine the free radical scavenging activity of fruits. On a w/w basis, higher total phenolics including flavonoids was found in achenes. The highest total anthocyanin content was found in the achenes of F. chiloensis and F. vesca, while F. ananassa presented higher antocyanin content in thalamus. The main anthocyanin in thalamus of F. ananassa (95%) were pelargonidin derivatives which were also present in F. chiloensis ssp. chiloensis f. patagonica (62.6%) but were not detected in F. vesca and F. chiloensis ssp. chiloensis f. chiloensis. In F. vesca and F. chiloensis ssp. chiloensis f. chiloensis only cyanidin derivatives were found both in thalamus and achenes.     

Antioxidant properties of aqueous extracts from Terminalia catappa leaves

Aqueous extracts were prepared from green, yellow fallen and red fallen leaves of Terminalia catappa L. (Combretaceae) and their antioxidant activity, scavenging and chelating abilities were evaluated. Aqueous extracts from three different leaves showed high antioxidant activities and moderate scavenging abilities on hydroxyl radicals at 1 mg/ml. EC₅₀ values in antioxidant activity were 0.549-0.557 mg/ml whereas those in scavenging ability on hydroxyl radicals were 0.631-0.686 mg/ml for aqueous extracts prepared from three leaves with boiling water for 3 min. EC50 values in reducing power were 0.15-0.23 mg/ml. EC50 values in scavenging abilities on superoxide anion and 1,1-diphenyl-2-picrylhydryl radicals were 0.36-0.44 and 10.4-35.3 mg/ml, respectively. EC50 values in chelating abilities on ferrous and cupric ions were 0.41-2.50 and 8.96-9.89 mg/ml, respectively. Based on the results obtained, the aqueous extracts displayed higher antioxidant properties. Six phenolic compounds identified in the aqueous were p-hydroxybenzoic acid, 4-hydroxyphenylpropionic acid, m-coumaric acid, 3,4-dihydroxybenzoic acid, p-coumaric acid and gallic acid. Therefore, the aqueous extracts from three different leaves of T. catappa might be a potential antioxidant supplement for application in food products or as a drink.     

Antioxidant and radical scavenging activities of the pyroligneous acid from a mangrove plant, Rhizophora apiculata

Total phenolics content, free radical scavenging activity, reducing power, and antioxidant activity of the pyroligenous acid from a mangrove plant, Rhizophora apiculata were evaluated. Dichloromethane extraction of the raw pyroligneous acid successfully yield 2 extracts, i.e. concentrated pyroligneous acid (CPA) and concentrated pyroligneous acid extract (CPAE). Phenolic contents in CPAE and CPA, expressed as (±)-catechin equivalents/g of the sample were 5465 ± 367 mg and 2502 ± 152 mg, and expressed as gallic acid equivalents/g of the sample were 2919 ± 209 mg and 1348 ± 90 mg, respectively. CPAE exhibited superior free radical scavenging activity with EC₅₀ value = 0.1235 mg/ml, or 80.96% of free radical scavenging capability. The ferric reducing power of CPAE was approximately 3.7, 5.1, 6.1, and 21.3 times higher than that of ascorbic acid, BHA, BHT and alpha-tocopherol. In phosphomolybdenum assay, CPAE showed the greatest antioxidant efficacy (A₆₉₅ = 1.278) compared to those of CPA and different standards. In addition, the free radical scavenging activity, ferric reducing power and total antioxidative activity of CPAE and CPA showed positive correlation with their total phenolic content with R² values ranging from 0.9624 to 0.9979.     

Antioxidant activity of Rhizoma Smilacis Glabrae extracts and its key constituent-astilbin

Rhizoma Smilacis Glabrae is widely consumed by Chinese as functional food and in folk medicine for its medicinal properties. In this study, methanol and water extracts of Rhizoma Smilacis Glabrae were prepared. The water extract was further divided into polysaccharide and supernatant fractions. Constituents in different extracts were analysed by capillary electrophoresis, and levels of total phenolics were also determined using the Folin-Ciocalteu method. Astilbin, the main constituent in the herb, was isolated and purified. Different antioxidant tests were employed to evaluate the antioxidant activities of the extracts and the isolated astilbin, and the results were compared with two commonly used synthetic antioxidants-butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). Methanol, water extract and supernatant fraction showed concentration dependent antioxidant activity while polysaccharide didn’t show any antioxidant activity. Purified astilbin showed the strongest antioxidant activity in comparison to any other extracts.     

Antioxidant properties of three extracts from Pleurotus citrinopileatus

Pleurotus citrinopileatus Sing. (Lentinaceae) was successfully cultivated and commercially available in Taiwan. The ethanolic, cold and hot water extracts were prepared from P. citrinopileatus fruit bodies, mycelia and fermentation filtrate and their antioxidant properties studied. For three samples, ethanolic extracts exhibited higher antioxidant activities than water extracts. Reducing powers of three extracts from fruit bodies were effective and 1.03-1.10 at 5 mg/ml. With regard to EC₅₀ values of scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl radicals, the effectiveness was in a descending order: ethanolic>hot water>cold water extracts. Scavenging abilities of water extracts from three samples on hydroxyl radicals were 53.4-80.1% at 20 mg/ml. Chelating abilities of cold and hot water extracts on ferrous ions were higher than those of ethanolic extracts. Contents of total phenols were in the descending order: fruit bodies (8.62-12.38 mg/g)>mycelia (5.84-7.85 mg/g)>filtrate (4.80-5.57 mg/g). Overall, three extracts from fruit bodies were more effective in antioxidant properties assayed than those from mycelia and filtrate. Ethanolic extracts were more effective in antioxidant properties assayed, except for scavenging abilities on hydroxyl radicals.     

Antioxidant properties of methanolic extracts from Agrocybe cylindracea

Agrocybe cylindracea (DC: Fr.) Mre. (Bolbitiaceae) was available in the form of fruit bodies, mycelia and fermentation filtrate. From these three forms, methanolic extracts were prepared and their antioxidant properties studied. The methanolic extract from fruit bodies showed high inhibitory ability on lipid oxidation (90.0-97.3%) at 5-20 mg/ml and had an EC₅₀ value of 0.94 mg extract/ml. EC₅₀ values in reducing power were 2.36, 6.89 and 5.00 mg extract/ml for fruit bodies, mycelia and filtrate, respectively. The scavenging ability of the methanolic extract from fruit bodies on 1,1-diphenyl-2-picrylhydrazyl radicals was 89.0% at 1 mg/ml whereas those from mycelia and filtrate was 91.4% and 94.9% at 10 mg/ml, respectively. The chelating abilities of methanolic extracts from fruit bodies, mycelia and filtrate on ferrous ions were 90.6%, 84.6% and 96.3% at 5 mg/ml, respectively. Total phenols were the major detected naturally occurring antioxidant components found in methanolic extracts from A. cylindracea and in the range of 15.55-23.47 mg/g. Based on EC₅₀ values, A. cylindracea was good in antioxidant properties, except for the scavenging ability on hydroxyl radicals.     

Antioxidant activity of microwave-assisted extract of longan (Dimocarpus Longan Lour.) peel

The longan (Dimocarpus Longan Lour.) peel was extracted with 95% ethanol employing microwave-assisted extraction and Soxhlet extraction method, the total phenolic content of microwave-assisted extract of Langan peel (MEL) and Soxhlet extract of Langan peel (SEL) reached 96.78 mg/g and 90.35 mg/g dry weight, respectively, expressed as pyrocatechol equivalents, which were quantified using Folin–Ciocalteu reagent. Subsequently, antioxidant properties of two extracts were investigated employing various established systems in vitro including 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, hydroxyl radical scavenging assay using a new resonance scattering (RS) method, reducing power and total antioxidant capacity. MEL and SEL showed excellent antioxidant in all test systems compared to synthetic antioxidant 2,6-di-ter-butyl-4-methylphenol (BHT) and the antioxidant activities of MEL were all superior to those of SEL. Furthermore, the suitability of MEL and SEL as substitute of BHT were determined in peanut oil, and the decrease of lipid oxidation were monitored using thiobarbituric acid-reactive substances (TBARS) assay. MEL and SEL treatment significantly (P < 0.05) reduced lipid oxidation in peanut oil compared to the control. No significant differences (P=0.05)$P=0.05)$ in lipid oxidation were detected between MEL, SEL and BHT samples of peanut oil.     

Antioxidant properties of hot water extracts from Agrocybe cylindracea

Agrocybe cylindracea (DC: Fr.) Mre. was available in the form of fruit bodies, mycelia and fermentation filtrate. From these three forms, hot-water extracts were prepared and their antioxidant properties were studied. Antioxidant activities of hot-water extracts from fruit bodies, mycelia and filtrate were 63.6%, 81.6% and 56.8% at 20 mg ml⁻¹, respectively. EC₅₀ values in reducing power were 2.72, 3.97 and 3.09 mg ml⁻¹ whereas those in scavenging abilities of 1,1-diphenyl-2-picrylhydrazyl radicals were 0.62, 1.66 and 0.82 mg ml⁻¹ for fruit bodies, mycelia and filtrate, respectively. At 20 mg ml⁻¹, the scavenging abilities of hydroxyl radicals were 80.1%, 57.0% and 54.3% for fruit bodies, mycelia and filtrate, respectively. With regard to EC₅₀ values in chelating abilities on ferrous ions, the hot-water extract from filtrate was better than that from mycelia. Total phenols were the major naturally occurring antioxidant components found in hot-water extracts and in the range of 23.74–30.16 mg g⁻¹. From EC₅₀ values obtained, it can be concluded that hot-water extracts from three forms of A. cylindracea were good in antioxidant properties.     

藜麦提取物体外抗氧化活性研究

以产自山西的藜麦为研究对象,以对DPPH·和ABTS+·的清除能力、铁还原力等为指标,评价了其提取物的体外抗氧化活性。结果表明,藜麦提取物对DPPH·和ABTS+·均有较强的清除能力,其IC₅₀分别为（43.13±2.41）mg/m L和（27.91±1.46）mg/m L;FRAP法测得藜麦抗氧化能力为（1.90±0.10）mg Fe SO₄/g;此外,藜麦提取物能够有效抑制自由基引发的亚油酸过氧化和牛血清白蛋白氧化降解。藜麦提取物具有较强的体外抗氧化活性,为其体内抗氧化活性的研究奠定了基础。