Phenolic composition,antioxidant and antimicrobial activities of free and bound phenolic extracts of Moringa oleifera seed flour

Phenolic compounds, antioxidant and antibacterial activities of defatted Moringa oleifera seed flour (DMF) were investigated. The total phenolic and flavonoid contents were measured using colorimetric methods. Free phenolic acid and flavonoid profiles were analyzed by high performance liquid chromatography, while antioxidant capacities were evaluated using scavenging assays of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), ferric reducing antioxidant power and total antioxidant capacity. The results showed that extractability of phenolic compounds was significantly higher (p < 0.05) in bound phenolic extract (4173.00 ± 32.22 mg gallic acid equivalents (GAE)/100 g) than in free phenolic extract (780.00 ± 14.2 mg GAE/100 g) and it showed higher antioxidant and antimicrobial activities. The IC₅₀ value for DPPH radical scavenging activity was 0.9 ± 0.05 and 14.9 ± 0.07 mg/mL for bound phenolic and free phenolic extracts, respectively. Bound phenolic extract was more effective (minimum inhibitory concentration (MIC), 0.06–0.157%) than free phenolic extract (MIC, 0.117–0.191%) against tested bacteria. Ten phenolic compounds (gallic acid, p-coumaric acid, ferulic acid, caffeic acid, protocatechuic acid, cinnamic acid, catechin, epicatechin, vanillin and quercetin) were identified and quantified in both extracts. These natural plant phenolics from Moringa seeds could be a good source of antioxidants and antibacterials for food and pharmaceutical industries.     

Phenolic and aroma compositions of pitomba fruit (Talisia esculenta Radlk.) assessed by LC–MS/MS and HS-SPME/GC–MS

Pitomba (Talisia esculenta Radlk.) is a Brazilian exotic fruit consumed specially in the Amazonian region. Because of its large consumption and also due to the lack of knowledge regarding its chemical composition, pitomba fruit was studied in relation to its phenolic and aroma constitution. Using liquid chromatography tandem mass spectrometry (LC–MS/MS), 13 phenolic compounds (catechins, flavonoids and organic acids) were tentatively identified by comparison with standards and by fragmentation patterns. A validated method was applied to quantify common phenolic compounds of the pitomba pulp, for which quinic acid was the main compound (507.8 ± 7.4 μg g^− 1 DWP). Gas chromatography mass spectrometry (GC–MS) was employed along with headspace solid-phase microextraction (HS-SPME) to assess the aroma composition of pitomba fruit. A total of 27 volatile organic compounds (VOCs) were tentatively identified for pitomba fruit, for which 2-phenethyl acetate (17.89%) and isopentyl acetate (13.43%) esters were the main VOCs, contributing to the characteristic aroma of pitomba. The antioxidant capacity of the extract of pitomba fruit was evaluated by ABTS, DPPH and ORAC assays. We observed that pitomba has a moderate antioxidant activity.     

Pilosocereus arrabidae (Byles & Rowley) of the Grumari sandbank,RJ, Brazil: Physical,chemical characterizations and antioxidant activities correlated to detection of flavonoids

The fruits of Pilosocereus arrabidae (Cactaceae) is found in the Grumari shoal, located in the Rio de Janeiro state, Brazil and, several studies have already highlighted the importance of consuming its fruits. This work aims to investigate the physical, mineral and physicochemical properties of its fruits as well as to establish the knowledge about their chemical constituents and antioxidant properties. The peel (Pe) and pulp (Pu) extracts were obtained by maceration with the following solvents: hexane (HX), dichloromethane (DCL), ethyl acetate (EAC) and ethanol 70% (ET). The extracts were analyzed by Gas Chromatography coupled to Mass Spectrometry (GC–MS) and, by High Performance Liquid Chromatography (HPLC–DAD and HPLC–MS) for its chemical investigation. For the antioxidant activity investigations the ORAC, DPPH and ferrous ion chelating capacity (FICC) tests were performed. As results, we found higher yields for peels (72%) compared to pulps (28%). By the physical–chemical analyses we point out the fruits as a good source of fiber (pulp: 6.01 mg/100 g; peel: 8.02 mg/100 g). The minerals were analyzed by the method of issuing flames and indicated high levels of selenium (DRI for pulp and peel: 147%) and manganese (DRI pulp: 97.69% and DRI peel: 269.56%). The total flavonoid contents of the fruits performed by HPLC–DAD presented 0.45 μg equiv. in quercetin/mL of peel EAC extract and 0.25 μg equiv. in rutin/mL of pulp EAC extract. The antioxidant activities by the ORAC, FICC and DPPH methods indicated that the ET extracts showed antioxidant activities above the standards adopted for the tests. Among these, we highlight the ET extract of the pulp with EC₅₀ of 17.57 ± 0.27 μg/mL, lower than Ginkgo biloba EGB761® (23.40 ± 0.04 mg/mL). By the FICC test the EAC extract of the peel and pulp showed 70.0% and 53.4% activity, respectively, at 500 mg/mL, higher than the standard quercetin (50.0%). By the HPLC–DAD and HPLC–MS methods there were detected, for the first time on this species, the presence of the following flavonoids on the EAC extracts: quercetin, rutin, catechin, dihydrokaempferol, quercetin 3 or 4′-O-glucoside, kaempferol and isorhamnetin. By the GC–MS analysis there were detected on the DCL extracts saturated fatty acids (palmitic and stearic) and, on the HX extracts, methylated sugars (peel) and menthol (pulp). To sum up, the fruits of P. arrabidae display antioxidant potential correlated to flavonoid presence, and, high levels of selenium, manganese and fibers, characteristics that can promote beneficial effects on human health.     

Citharexylum solanaceum fruit extracts: Profiles of phenolic compounds and carotenoids and their relation with ROS and RNS scavenging capacities

Citharexylum solanaceum is a native fruit from Brazil, which both bioactive compounds and antioxidant potential were not yet investigated. Thus, the freeze-dried extracts of seed and pulp + skin of C. solanaceum fruits were obtained after solid-liquid extraction with ethanol and their bioactive compounds composition, namely phenolic compounds and carotenoids, were determined. The antioxidant capacity of both extracts against physiologically relevant reactive oxygen species (ROS) and reactive nitrogen species (RNS) was further investigated. Both C. solanaceum extracts showed high contents of phenolic compounds; however, pulp + skin extract presented 2.4-times more phenolic compounds (33.54 mg/g) than the seed extract (14.09 mg/g). Verbascoside (phenylpropanoid) was the major compound identified in both extracts (11–25 mg/g). Regarding the carotenoid composition, all-trans-lutein (14–42 μg/g) and all-trans-β-carotene (13–44 μg/g) were the major compounds in both extracts. The high content of phenolic compounds and carotenoids in pulp + skin extract might explain its higher scavenging capacity against all the ROS/RNS as compared to seed extract. In general, both extracts showed better scavenging capacity for the RNS than for the ROS. Our results indicate that C. solanaceum fruits can be explored as an important natural source of antioxidant compounds.     

Retention of Physicochemical and Antioxidant Properties of Dehydrated Bael (Aegle Marmelos) and Palmyra (Borassus Flabellifer) Fruit Powders

Present study was carried out to evaluate the effect of various dehydration techniques such as sun drying, solar drying, drying after freezing (Freeze for one hour followed by mechanical drying at 55 °C), vacuum drying and drying using lab scale air oven on proximate composition and retention of antioxidants in different fruit powder prepared from Bael (Aegle marmelos) and Palmyra (Borassus flabellifer). Moisture content, Total Ash, Crude fiber %, Fat %, Crude protein %, total phenolic content,β –Carotene and antioxidant activity were tested. The antioxidant activity was measured based on the ability of fruit extract to scavenge 1, 1- diphenyl-2-picrylhydrazyl (DPPH). Among different drying treatments the highest fat percentage recorded by the solar dried palmyra fruit powder and there is no significant difference (α= 0.05) between sun drying and vacuumed drying. Higher concentration of β -Carotene and total phenolic content were recorded in vacuum dried samples both in bael and Palmyra fruit powders and it significantly different (α= 0.05) from other treatments. The scavenging activity of bael fruit powder in vacuum drying was ranged from 65.36% to 81.33% of the concentration 200 μg/ml to1000 μg/ml and the palmyra fruit powder was recorded 57.32% to 83.25% of the concentration 200 μg/ml to1000 μg/ml. Vacuum dried fruit powders of palmyra and bael were given highest radical scavenging activity and the scavenging activity of palmyra fruit powder is higher than the bael. Therefore vacuum drying can be recommended as the most effective drying method to protect chemical characteristics and retention of antioxidant properties of fruit powders.     

Sequential high pressure extractions applied to recover piceatannol and scirpusin B from passion fruit bagasse

Passion fruit seeds are currently discarded on the pulp processing but are known for their high piceatannol and scirpusin B contents. Using pressurized liquid extraction (PLE), these highly valuable phenolic compounds were efficiently extracted from defatted passion fruit bagasse (DPFB). PLE was performed using mixtures of ethanol and water (50 to 100% ethanol, w/w) as solvent, temperatures from 50 to 70 °C and pressure at 10 MPa. The extraction methods were compared in terms of the global yield, total phenolic content (TPC), piceatannol content and the antioxidant capacity of the extracts. The DPFB extracts were also compared with those from non-defatted passion fruit bagasse (nDPFB). Identification and quantification of piceatannol were performed using UHPLC–MS/MS. The results showed that high TPC and piceatannol content were achieved for the extracts obtained from DPFB through PLE at 70 °C and using 50 and 75% ethanol as the solvent. The best PLE conditions for TPC (70 °C, 75% ethanol) resulted in 55.237 mg GAE/g dried and defatted bagasse, whereas PLE at 70 °C and 50% ethanol achieved 18.590 mg of piceatannol/g dried and defatted bagasse, and such yields were significantly higher than those obtained using conventional extraction techniques. The antioxidant capacity assays showed high correlation with the TPC (r > 0.886) and piceatannol (r > 0.772). The passion fruit bagasse has therefore proved to be a rich source of piceatannol and PLE showed high efficiency to recover phenolic compounds from defatted passion fruit bagasse.     

Use of natural food/plant extracts: cloudberry (Rubus Chamaemorus), beetroot (Beta Vulgaris “Vulgaris”) or willow herb (Epilobium angustifolium) to reduce lipid oxidation of cooked pork patties

The antioxidant activity of plant extracts (100 and 500 mg/kg) from cloudberry, willow herb and beetroot on thiobarbituric acid (TBA) reactive substances (TBARS) development and hexanal content of cooked pork patties was investigated. Pure quercetin, rutin and caffeic acid were tested in parallel for comparison. The most potent antioxidants on stabilizing oxidation were cloudberry extract and quercetin. The lowest antioxidant activity was found with the addition of pure rutin to the meat. Caffeic acid showed an intermediate activity. At a concentration of 100 mg/kg, beetroot and willow herb extracts showed an antioxidant activity on TBARS and hexanal contents similar to that observed for caffeic acid during 3 days of refrigerated display, while cloudberry extract was as potent as quercetin. At a concentration of 500 mg/kg, beetroot and willow herb extract stabilized hexanal production of cooked pork patties after 6 days of refrigerated storage in a way comparable to that observed for pure quercetin and cloudberry extract. TBARS numbers were well correlated with hexanal content in cooked pork patties on day 3 of refrigerated storage. However, hexanal production and TBARS numbers were not highly correlated in samples with the highest level of beetroot (500 mg/kg). Hexanal production was inhibited by the high level of beetroot, but TBARS production was not, perhaps because the red color of beetroot extract interfered with the determination of the pink TBA chromogen.     

Antioxidant and type II diabetes related enzyme inhibition properties of methanolic extract of an underutilized food legume,Canavalia ensiformis (L.) DC: Effect of traditional processing methods

The methanolic extract of Canavalia ensiformis (L.) DC (Jack bean) seed, an underutilized food legume collected from India was analyzed for antioxidant and health relevant functional properties. The raw seeds contained total free phenolic content of 12.98 ± 1.63 g catechin equivalent/100 g extract DM. The raw seed extract exhibited ferric reducing/antioxidant power (FRAP, 1218 mmol Fe[II]/mg extract), inhibition of β-carotene degradation (49.81%), radical scavenging activity against DPPH (56.78%) and superoxide (35.89%). In addition, 77.56% of α-amylase and 75.45% of α-glucosidase enzyme inhibition characteristics were found under in vitro starch digestion bioassay. Sprouting + oil-frying caused an apparent increase on the total free phenolic content with significant improvement on antioxidant and free-radical scavenging capacity, while soaking + cooking as well as open-pan roasting treatments showed diminishing effects. Inhibition of α-amylase and α-glucosidase enzyme activities were declined to 22.69 and 42.69%, respectively during sprouting + oil-frying treatment is more desirable for the dietary management of type II diabetic patients.     

Influence of extraction process on antioxidant capacity of spent coffee

Spent coffee that is produced in tons by restaurants and cafeterias, and consumers at domestic levels, could be a good opportunity to have an important source of natural antioxidants. The main aim of this work was to study the influence of several process factors on the antioxidant capacity extraction from spent coffee. Total phenolic compounds, radical scavenging activity (ABTS and DPPH) and browned compounds (Abs 420 nm) of spent coffee extracts obtained with continuous (Soxhlet 1 h and 3 h) and discontinuous methods (solid–liquid extraction and filter coffeemaker), several solvents (water, ethanol, methanol and their mixtures), successive extractions, and water with different pHs (4.5, 7.0 and 9.5) were carried out. Spent coffee extracts with the highest antioxidant capacity were obtained after one extraction with neutral water (pH 7.0) in a filter coffeemaker (24 g spent coffee per 400 mL water). Furthermore, spent coffee defatting and extract lyophilization allowed us to obtain spent coffee extracts powder with high antioxidant capacity that can be used as an ingredient or additive in food industry with potential preservation and functional properties.     

Effect of Firm Ripe Plantain Fruit Flour Addition on the Chemical,Sensory and Microbial Quality of Fura Powder

This study was carried out to determine the effect of firm ripe plantain fruit flour addition on the chemical, sensory and microbial quality of fura powder. Millet flour was supplemented with plantain flour at substitution levels of 0, 10, 20, 30 and 40 to obtain fura powder. The chemical composition, sensory properties and microbial quality were determined using standard methods of analysis. Addition of firm ripe plantain fruit flour significantly (p < 0.05) increased vitamin A, vitamin C, potassium, fibre and soluble solids (°Brix) levels of plantain-fura with the increasing level of plantain flour substitution. Fat content decreased from 6.0% to 3.0% with increased levels of plantain fruit flour addition. Similar trend was observed in protein content as it decreased from 13.0% to 10.0%. Microbial analysis results during three months of storage showed significant (p > 0.05) decrease in the microbial population with the increasing level of plantain flour addition. The results showed that blending of firm ripe plantain flour with millet flour would produce fura that is shelf stable, rich in natural anti-oxidant vitamins and safe for consumption.     

Evaluation of the antioxidant,antiproliferative and antimutagenic potential of araçá-boi fruit (Eugenia stipitata Mc Vaugh — Myrtaceae) of the Brazilian Amazon Forest

Eugenia stipitata is a fruit from Amazonia rich in terpene, volatile compounds, fiber, and vitamin C. The fruit is recognized for its high antioxidant activity and has attracted much attention due to their potential health benefits to humans. The total polyphenols, antioxidant, antiproliferative, antimutagenic and antigenotoxic activities of E. stipitata ethanolic extract were investigated. Total polyphenols were determined by the Folin–Ciocalteu method and showed 184.05 ± 8.25 mg GAE/100 g. The radical scavenging activity was DPPH _IC50 0.69 ± 0.23 μg/mL and TAC-ORAC_FL 371.98 μmol.TE/100 g. The extract was evaluated for its ability to inhibit the growth of tumor cell lines and had not complete cystostatic effect against any of the tested cell lines. Antimutagenic and antigenotoxic activities were investigated by micronucleus test and comet assay in mice, respectively. Ethanolic extract of E. stipitata showed higher antimutagenic and antigenotoxic properties at the highest concentration tested (300 mg/kg of body weight). In conclusion, these results suggest that this fruit could be used as a preventive agent against cancer.     

Tea and herbal infusions: Their antioxidant activity and phenolic profile

Tea and herbal infusions have been studied for their polyphenolic content, antioxidant activity and phenolic profile. The total phenolics recovered by ethyl acetate from the water extract, were determined by the Folin–Ciocalteu procedure and ranged from 88.1 ± 0.42 (Greek mountain tea) to 1216 ± 32.0 mg (Chinese green tea) GAE (Gallic acid equivalents)/cup. The antioxidant activity was evaluated by two methods, DPPH and chemiluminescence assays, using Trolox and quercetin as standards. The EC₅₀ of herbal extracts ranged from 0.151 ± 0.002 mg extract/mg DPPH (0.38 quercetin equivalents and 0.57 Trolox equivalents), for Chinese green tea, to 0.77 ± 0.012 mg extract/mg DPPH (0.08 quercetin equivalents and 0.13 Trolox equivalents), for Greek mountain tea. Chemiluminescence assay results showed that the IC₅₀ ranged from 0.17 ± 3.4 × 10⁻³ μg extract/ml of the final solution in the measuring cell (1.89 quercetin and 5.89 Trolox equivalents) for Chinese green tea, to 1.10 ± 1.86 × 10⁻² g extract/ml of the final solution in the measuring cell (0.29 quercetin and 0.90 Trolox equivalents) for Greek mountain tea. The phenolic profile in the herbal infusions was investigated by LC-DAD-MS in the positive electrospray ionization (ESI⁺) mode. About 60 different flavonoids, phenolic acids and their derivatives have been identified.     

Effect of Fermentation Time on the Phenolic,Flavonoid and Vitamin C Contents and Antioxidant Activities of Okra (Abelmoschus esculentus) Seeds

The objective of this study was to evaluate the effect of fermentation on the phenolic, vitamin C and total flavonoid contents and antioxidant properties of okra seed. The okra seeds were removed from the pod of matured and fibrous okra that cannot be easily cut with kitchen knife, and was allowed to undergo chance fermentation for 120 h. Samples were taken for antioxidant analysis at 24 h, 72 h and 120 h, respectively. The aqueous extracts of fermented and unfermented okra seeds were obtained and subsequently used for the analysis. The phenolic, vitamin C and total flavonoid contents and the antioxidant properties (ferric reducing antioxidant power (FRAP) and 1, 1-diphenyl-2 picrylhydrazyl (DPPH) free radical-scavenging ability) of the extracts were determined. The results revealed that fermented okra seeds had significantly (p < 0.05) higher phenolic content, vitamin C, total flavonoid and non-flavonoid contents and showed greater antioxidant activities than unfermented okra seed. Okra seeds fermented for 24 h exhibited the highest ferric reducing antioxidant power of 980 mgAAE/100 g and the least IC₅₀ of 2.27 mg/ ml for DPPH free radical-scavenging ability than the others. Okra seeds fermented for 24 hours had significantly (p < 0.05) higher antioxidant activities. The product from fermented okra seeds could be used for the production of functional foods.     

Phenolic profiles and antioxidant activity of litchi pulp of different cultivars cultivated in Southern China

The phenolic profiles and antioxidant activity of litchi pulp of 13 varieties were investigated. The free, bound and total phenolic contents were 66.17–226.03, 11.18–40.54, and 101.51–259.18 mg of gallic acid equivalents/100 g, respectively. The free, bound and total flavonoid contents were 16.68–110.33, 10.48–22.75, and 39.43–129.86 mg of catechin equivalents/100 g, respectively. Free phenolics and flavonoids contributed averagely 80.1% and 75% to their total contents, respectively. Six individual phenolics (gallic acid, chlorogenic acid, (+)-catechin, caffeic acid, (?)-epicatechin, and rutin) were detected in litchi pulp by HPLC. The contents of each compound in free and bound fractions were determined. Significant varietal discrepancy in antioxidant activity was also found by FRAP and DPPH scavenging capacity methods. Antioxidant activity was significantly correlated with phenolic and flavonoid contents. Thus, phenolics and flavonoids exist mainly in the free form in litchi pulp. There were significant varietal differences in phytochemical contents and antioxidant activity of litchi pulp.     

Antioxidant Activity of Loquat (Eriobotrya japonica Lindl.) Fruit Peel and Pulp Extracts in Stabilization of Soybean Oil During Storage Conditions

The antioxidation activities of Eriobotrya japonica (Lindl.) fruit peel and pulp extracts were determined using DPPH, β-carotene, and Rancimat methods. Results showed that ethanol-water extract of peel and ethanol extract of pulp had the highest antioxidant activity. Protective effects of extracts in stabilization of soybean oil were tested and compared to tert-butyl hydroquinone by measuring peroxide values, free fatty acids, thiobarbituric acid, oxidative stability, and conjugated dienes and trienes values during storage (65 days at 25°C). Results showed that the ethanol-water extract of peel at 400 ppm exhibited stronger antioxidant activity, but the highest effect was observed in tert-butyl hydroquinone.     

Antioxidant activity of Bupleurum kaoi Liu (Chao et Chuang) fractions fractionated by supercritical CO2

Bupleurum kaoi Liu was mixed with ethanol at a ratio of 1:4 (v/w) for 24 h to yield ethanol extract. Extract was further fractionated using supercritical CO₂ (SC-CO₂) under the following operating conditions: 40°C and a pressure of 20, 15, 10 or 5 MPa into R, F1, F2 or F3 fractions, respectively. To assess the selectivity of the fractionation, four fractions were characterized in terms of total phenol contents, the antioxidant abilities and the antioxidant mechanisms. Experimental results indicated that fractionation altered the composition distributions and the antioxidant activities, including antioxidant ability, reducing power and the scavenging capacity of DPPH, superoxide, and hydroxyl radicals. The antioxidant activities increased with fraction concentrations. The scavenging effect on DPPH of B. kaoi L. fractions at 10 g/l was R (53%), F1 (65%), F2 (71%), and F3 (76%), respectively. At a concentration of 5 g/l, all fractions can inhibit the O₂·^—formation by over 50%. Fractions R, F2 and F3 at concentrations of 3 g/l can trap over 50% of the OH groups. Notably, this in vitro study of antioxidant effects demonstrated that antioxidant activities were correlated well with the contents of phenol compounds. F3 fraction, contained the highest levels of total phenol contents, was the best inhibitor of lipid peroxidation and scavengers of DPPH, O₂·^— and ·OH radicals.     

Extraction efficiency and validation of an HPLC method for flavonoid analysis in peppers

Efficient extraction procedures including solvent selection, sample to solvent ratio, extraction time, and hydrolysis condition were developed for the analysis of flavonoids in peppers. Maximum flavonoids were extracted in ethanol, while N-N-dimethylformamide was able to extract myricetin, which was rarely reported in peppers. Flavonoids were obtained at the 1:8 ratio of sample to solvent after 3 h of extraction time. The effective hydrolysis of glycosides to aglycones was observed in 3 M HCl at 60 min at 95 °C. The validated HPLC method was applied to quantify five flavonoids from peppers. The ethanol extract exhibited maximum total phenolics (1.39 mg of catechin equivalent/g), and the DPPH assay showed a similar free radical scavenging activity in MeOH, EtOH, and EtOH:water (80:20). Reducing property of the MeOH extract was higher than the EtOH extract. This study demonstrated that the developed methods for the extraction of pepper flavonoids could be useful for analysing large numbers of samples.     

Evaluation and improvement of antioxidant and antibacterial activities of supercritical extracts from clove buds

For the first time, extracts from clove buds obtained by supercritical carbon dioxide extraction were screened for antioxidant and antibacterial activities. Additionally, antioxidant and antibacterial activities of extracts obtained by the supercritical extraction of the clove bud–oregano leaf mixtures were studied. Supercritical extract of pure clove had the highest eugenol (64%) and total phenolic content (530.56 mg GAE/g_extract). All extracts had antioxidant activity comparable to synthetic antioxidants against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and formation of peroxides. Presence of 0.6% and 5% of oregano extract in the clove extracts obtained from the clove–oregano plant mixtures improved their antioxidant activity with respect to the extract from pure clove. Clove extract showed moderate antibacterial activities against selected Staphylococcus and Enterococcus bacterial strains. Presence of 50% of the oregano extract improved antibacterial activity of clove extract against all tested strains and resulted in a synergistic antibacterial activity against Methicillin-resistant Staphylococcus haemolyticus strain (MIC ? 1.25 μg/mL). Study demonstrated great potential of supercritical clove extract as natural functional ingredient and the possibility of increasing its antioxidant and antibacterial efficiencies in order to apply lower concentrations and to reduce undesirable flavour notes and toxicological effects in final products.     

Antioxidant properties of hot water extracts from Ganoderma tsugae Murrill

Ganoderma tsugae Murrill (Ganodermataceae) were available in the form of mature and baby Ling chih, mycelia and fermentation filtrate. From these four forms, hot water extracts were prepared and their antioxidant properties were studied. Hot water extracts from mature and baby Ling chih showed high antioxidant activities (78.5% and 78.2%) at 20 mg/ml, and had EC₅₀ values of 7.25 and 5.89 mg extract/ml, respectively. EC₅₀ values in reducing power were 1.12, 1.37, 2.48 and 1.41 mg extract/ml, whereas those in scavenging abilities of 1,1-diphenyl-2-picrylhydrazyl radicals were 0.30, 0.40, 0.72 and 5.00 mg extract/ml for Ling chih, baby Ling chih, mycelia and filtrate, respectively. At 20 mg/ml, scavenging abilities on hydroxyl radicals were in the descending order of Ling chih>baby Ling chih>mycelia>filtrate. Total phenols were the major naturally occurring antioxidant components found in hot water extracts and in the range of 40.86–42.34 mg/g. From EC₅₀ values obtained, fruit bodies of G. tsugae (Ling chih and baby Ling chih) were good in antioxidant properties, except for the chelating ability on ferrous ions.     

Antioxidant activity and characterization of constituents in copao fruits (Eulychnia acida Phil., Cactaceae) by HPLC–DAD–MS/MSn

Copao (Eulychnia acida Phil., Cactaceae) is an endemic species occurring in arid areas of northern Chile. The fruits are commercialized by peasants within the Elqui and Limari valleys and are appreciated for its acidic and refreshing taste. We now report the total phenolic (TP) and total flavonoid (TF) content, antioxidant activity, phenolic composition and main phenolic distribution in pulp and epicarp of copao fruits from different harvesting places from both valleys. The ascorbic acid content was determined in fresh fruit pulp, epicarp and juice. The phenolic-enriched extract was analyzed for antioxidant effect and composition. Ferulic acid, 9,10-dihydroxy-4,7-megastigmadien-3-one hexoside, isorhamnetin and quercetin glycosides were identified by HPLC–DAD–MS/MS analysis. The main compounds were isolated and fully characterized by NMR techniques. The main phenolic in the samples was isorhamnetin-3-O-[α-rhamnopyranosyl-(1 → 6)-β-glucopyranoside]. The HPLC pattern of the phenolic-enriched extracts of the fruits allows a differentiation of samples from the Elqui and Limari valleys. All fruit extracts and Amberlite-retained fraction from the methanolic extract were devoid of toxicity against human gastric AGS cells and human lung fibroblasts, with IC₅₀ values > 400 μg/mL for AGS and 344 to > 400 μg/mL for fibroblasts, respectively. The compound identification, associated with the antioxidant activity and insignificant cell toxicity, adds relevant information for the possible development of this native fruit into a new crop.