Effect of Fertilization on Phenolics of Rapeseeds and Their Antioxidant Potential.

Three varieties of rapeseed (Castilla, California, and Nelson F1) were cultivated using medium-intensive (control), intensive, and economical (spare) technologies with different nitrogen and sulfur fertilization techniques. The antioxidant potential of rapeseeds was investigated using ABTS, FRAP, and DPPH assays. The content of total phenolic compounds was determined using the Folin-Ciocalteu phenol reagent. The profile of phenolic compounds was determined using high-performance liquid chromatography (HPLC). Diversifying fertilization in various ways influenced the content of phenolic compounds in extracts of rapeseed. In extracts from the Nelson F1 rapeseeds, intensive cultivation resulted in a lower content of phenolic compounds compared to the control group. Economic fertilization reduced the content of phenolic compounds in seeds from the California variety. HPLC chromatograms of the extracts were characterized by the presence of five (California and Castilla) and six (Nelson F1) main phenolic compounds. Two compounds were identified as sinapine and sinapic acid; others were classified as derivatives of sinapic acid. The effect of fertilization on the antioxidant activity of the seeds and their extracts varied depending on the plant variety and antioxidant assay. For the Castilla and California varieties, no differences were found in the results of the ABTS assay. The antiradical activity against ABTS•+ of extracts from the Nelson F1 intensive and spare cultivated seeds was higher than that of extracts from control seeds. The FRAP values of extracts/seeds from the Castilla variety cultivated using different methods did not differ significantly. The results of the DPPH assay were not affected by fertilization in the case of extracts from the California and Castilla varieties. However, the extracts from spare cultivated seeds of Nelson F1 exhibited stronger antiradical activity against DPPH•. These findings highlight the complex relationship between fertilization practices, phenolic compound accumulation, and antioxidant activity in rapeseed. Integrating varietal traits and cultivation practices is crucial for optimizing the nutritional benefits of rapeseed.

Phenolic composition and correlation with antioxidant properties of various organic fractions from Hertia cheirifolia extracts.

Hertia cheirifolia L. is a medicinal plant that has been used for a long time in folk Mediterranean medicine. The aim of the present study was to analyze and compare the phenolic profile and the antioxidant potential of organic fractions from H. cheirifolia extracts. Crude methanolic extracts were firstly prepared from the different parts of the plant. Then four different organic fractions were obtained by fractioning each extract, using different solvents with increasing polarity (hexane, chloroform, and ethyl acetate). The Phenolic content was analyzed using a UV-Vis colorimetric methods followed by a qualitative and quantitative analysis by high performance liquid chromatography-diode array detector (HPLC-DAD) system. After that, the antioxidant potential of the different organic fractions was evaluated using DPPH and ABTS free radical scavenging assays, reducing power of iron (FRAP) and inhibition of ß-carotene oxidation tests. Our results revealed that ethyl acetate fractions (EA) contained the highest content of total phenolics (100-250 mg GAE/g). Indeed, the ethyl acetate fraction from the flower extract (EA-F) displayed the lowest IC50 values for the scavenging of DPPH and ABTS free radicals (38.83 ± 0.34 µg/ml and 23.76 ± 0.11 µg/ml, respectively). Also, the strongest iron reducing power (2628.87 ± 16.47 µmol Fe2+Eq/ml) and the best rate of inhibition of the ß-carotene oxidation (58.91 ± 5.79 %) were recorded. In sum, the present study suggests that, the organic fractions from H. cherifolia are potential natural antioxidants and this is probably related to their phenolics content and structure.

Characterization of Antioxidant and Antimicrobial Activity and Phenolic Compound Profile of Extracts from Seeds of Different Vitis Species.

Seeds of Vitis vinifera L. with a high content of bioactive compounds are valuable by-products from grape processing. However, little is known about the bioactivity of seeds from other Vitis species. The aim of this study has been to compare the phenolic composition, antimicrobial activity, and antioxidant activity of extracts from seeds of four Vitis species (V. riparia Michx., V. californica Benth., V. amurensis Rupr., and V. vinifera L.). Antioxidant activities were assessed as ferric-reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) scavenging activity, and oxygen radical absorbance capacity (ORAC). The antimicrobial activity was determined using the microdilution method against some Gram-negative (Escherichia coli, Salmonella enterica ser. Typhimurium, and Enterobacter aerogenes) and Gram-positive (Enterococcus faecalis and Staphylococcus aureus) bacteria. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to evaluate the phenolic profile of extracts. Flavan-3-ols, procyanidins, phenolic acids, flavonols, anthocyanins, and stilbenoids were detected. (+)-Catechin and (-)-epicatechin turned out to be the most abundant in the phenolic profile of V. amurensis seed extract. Phenolic acids prevailed in the extract from V. vinifera seeds. The V. riparia and V. californica seed extracts had higher contents of most individual phenolics compared to the other Vitis species. They also showed a higher total phenolic content, DPPH• scavenging activity, ORAC, and overall antibacterial activity. Total phenolic content significantly correlated with antioxidant activity and antimicrobial activity against E. coli. The principal component analysis (PCA) showed discrimination between V. vinifera, V. amurensis, and clustered V. riparia and V. californica with respect to variables. To recapitulate, this research demonstrates that seeds of different Vitis species, especially V. riparia and V. californica, are sources of molecules with antioxidant and antimicrobial activities that can be used in different sectors, such as in the food, cosmetic, and pharmaceutical industries.

Phenolic Compound Profile and Antioxidant Capacity of Flax (Linum usitatissimum L.) Harvested at Different Growth Stages.

The profile of phenolic compounds changes during the growth of a plant and this change affects its antioxidant potential. The aim of this research has been to find the growth stage of flax with the highest antioxidant capacity, and to determine the phenolic compounds responsible for such a capacity. Flax was harvested in six growth stages: from stem extension to mature seeds. The phenolic compounds were identified using LC-TOF-MS and quantified in an extract and in the fresh matter (FM) of each growth stage. The radical scavenging activity against ABTS•+ and DPPH•, the ferric-reducing antioxidant power (FRAP), and the antioxidant activity in the ß-carotene-linoleic acid emulsion system were determined. Mono- and di-C-glycosyl flavones were found to be the most abundant phenolics of the aerial parts of flax, which also showed the highest content of isoorientin (210-538 µg/g FM). Coniferin, its derivative, and hydroxycinnamic acid derivatives were also detected. The plant was richer in flavone C-glycosides from stem extension to seed ripening (1105-1413 µg/g FM) than at the mature seed stage (557 µg/g FM). Most of the individual flavone C-glycoside contents in the extracts decreased when increasingly older plants were considered; however, the isoorientin content did not change significantly from the steam extension to the seed ripening stages. The antiradical activity against ABTS•+ and FRAP was higher for the aerial parts of the flax harvested at the flowering, brown capsule, and seed ripening stages, mainly due to the presence of flavone C-glycosides. The oxidation of ß-carotene-linoleic acid emulsion was instead inhibited more effectively by the extracts from plants at the brown capsule and mature seed stages. Coniferin and its derivative were significantly involved in this activity. The extracts from the aerial parts of the flax harvested from flowering to seed ripening could be a valuable source of flavone C-glycosides for use as nutraceuticals and components of functional foods.

Synergistic, additive, and antagonistic antioxidant effects in the mixtures of curcumin with (-)-epicatechin and with a green tea fraction containing (-)-epicatechin.

The combinations of curcumin with green tea flavan-3-ols produce various synergistic biological effects. This study aimed to verify the antioxidant effects in mixtures of curcumin with (-)-epicatechin (EC) or with EC fraction from green tea in a non-polar lipid system (triacylglycerol autoxidation) and in a polar conditions (ABTS assay). Curcumin was 2.5-2.6 and 2.9-3.6 times weaker antioxidant than EC and EC fraction, respectively. The synergism was found in mixtures using the isobologram analysis of ABTS•+ scavenging activity results. The strongest effect with a combination index of 0.751 was in the equimolar mixture of pure compounds. In the lipid system, antagonism occurred for curcumin and EC fraction combination. However, an additive effect was found between curcumin and EC. In conclusion, the antioxidant effects in the curcumin and EC mixtures depended on the polarity of the assay media, the ratio of antioxidants, and presence other phenolics in the system.

Sunflower (Helianthus annuus L.) Plants at Various Growth Stages Subjected to Extraction-Comparison of the Antioxidant Activity and Phenolic Profile.

The aim of this study was to evaluate the differences in the antioxidant activity and phenolic profile of sunflower (Helianthus annuus L.) extracts obtained from the aerial parts of plants harvested at five growth stages. In vitro assays were used to determine the antioxidant activity, i.e., ABTS•+ and DPPH• scavenging activity, the ferric-reducing antioxidant power (FRAP) and the ability to inhibit ß-carotene-linoleic acid emulsion oxidation. Phenolic compounds, such as mono- and dicaffeoylquinic acid isomers and caffeic acid hexose, were identified using the LC-TOF-MS/MS technique. The predominant compound during the growth cycle of the plant was 3,5-di-O-caffeoylquinic acid, whose content was the highest at the mid-flowering stage. The total phenolic content was also the highest in sunflowers at the mid-flowering stage. The main phenolic compound contents were closely correlated with ABTS•+ and DPPH• scavenging activity and FRAP. No significant correlation was found between the total phenolic content and the antioxidant activity in the emulsion system. The highest antiradical activity and FRAP were generally determined in older plants (mid-flowering and late flowering stages). In conclusion, the aerial parts of sunflowers, in particular those harvested at the mid-flowering stage, are a good plant material from which to obtain phenolic compound extracts, albeit mainly of one class (esters of caffeic acid and quinic acid), with high antioxidant activity.

Comparison of Oxidative Status of Human Milk, Human Milk Fortifiers and Preterm Infant Formulas.

Preterm and low birth weight infants require specific nutrition to overcome the accumulated growth deficit, and to prevent morbidities related to postnatal growth failure. In order to guarantee an adequate nutrient-intake, mother's own milk, when available, or donor human milk, are usually fortified with additional nutrients, in particular proteins. Fortification with processed ingredients may result in additional intake in oxidative compounds, deriving from extensive heat treatments, that are applied during processing. The aim of the present work was to compare the in vitro antioxidant activity and oxidative compound content conveyed by different preterm infant foods and fortifiers, namely raw and pasteurized human milk, two different preterm infant formulas, three bovine milk-based fortifiers and two experimental donkey milk-based fortifiers. Univariate and multivariate statistical analyses revealed significant differences between the different products. The use of human milk minimizes the intake of dietary oxidative compound in comparison to infant formulas, irrespective of pasteurization or fortification, especially as far as malondialdehyde content is concerned. The addition of fortifiers to human milk increases its antioxidant capacity, and the choice of the protein source (hydrolysed vs. whole proteins) differently impacted the resulting total antioxidant capacity of the diet.

Antioxidant Activity and Phenolic Composition of Amaranth (Amaranthus caudatus) during Plant Growth.

The antioxidant activity and phenolic composition of the aerial part of Amaranthus caudatus at seven stages of development were investigated. Total phenolic content, ABTS•+, DPPH•, and O2•- scavenging activity, ferric-reducing antioxidant power (FRAP), and Fe2+ chelating ability were evaluated. The phenolic profile was characterized by 17 compounds. Rutin was predominant in all growth stages, although its content, similar to the quantity of other phenolics, changed during the growth cycle. Flavonols were most abundant in the plants of early flowering and grain fill stages. In contrast, the highest content of hydroxycinnamic acid derivatives was found in the early vegetative stage. The results of antioxidant assays also showed significant differences among plant stages. Generally, the lowest antioxidant activity was found in the shooting and budding stages. Significantly higher activity was observed in amaranths in earlier (vegetative) and later (early flowering and grain fill) stages, suggesting that plants in these stages are valuable sources of antioxidants.

Genotype-Related Differences in the Phenolic Compound Profile and Antioxidant Activity of Extracts from Olive (Olea europaea L.) Leaves.

The phenolic compound contents and antioxidant activities of the leaf extracts of nine olive genotypes were determined, and the obtained data were analysed using chemometric techniques. In the crude extracts, 12 compounds belonging to the secoiridoids, phenylethanoids, and flavonoids were identified. Oleuropein was the primary component for all genotypes, exhibiting a content of 21.0 to 98.0 mg/g extract. Hydroxytyrosol, verbascoside, luteolin 7-O-glucoside, and luteolin 4'-O-glucoside were also present in noticeable quantities. Genotypes differed to the greatest extent in the content of verbascoside (0.45⁻21.07 mg/g extract). The content of hydroxytyrosol ranged from 1.33 to 4.03 mg/g extract, and the aforementioned luteolin glucosides were present at 1.58⁻8.67 mg/g extract. The total phenolic content (TPC), DPPH• and ABTS•+ scavenging activities, ferric reducing antioxidant power (FRAP), and ability to inhibit the oxidation of -carotene-linoleic acid emulsion also varied significantly among genotypes. A hierarchical cluster analysis enabled the division of genotypes into three clusters with similarity above 60% in each group. GGE biplot analysis showed olive genotypes variability with respect to phenolic compound contents and antioxidant activities. Significant correlations among TPC, FRAP, the values of both radical scavenging assays, and the content of oleuropein were found. The contents of 7-O-glucoside and 4'-O-glucoside correlated with TPC, TEAC, FRAP, and the results of the emulsion oxidation assay.

Antioxidant Potential of Grass Pea Seeds from European Countries.

Phenolic compounds were extracted from seeds of 30 varieties of grass pea (Lathyrus sativus) into 80% (v/v) methanol. The total phenolics compounds content of the extracts and their antioxidant activity were determined using Folin-Ciocalteu's phenol reagent and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric-reducing antioxidant power (FRAP) methods, respectively. Total phenolic contents ranged from 1.88 to 7.12 mg/g extract and 20.3 to 70.3 mg/100 g seeds. The extracts and seeds were characterized using Trolox equivalent antioxidant capacity values of 0.015⁻0.037 mmol Trolox/g extract and 0.158⁻0.372 mmol Trolox/100 g seeds, and FRAP values of 0.045⁻0.120 mmol Fe2+/g extract and 0.487⁻1.189 Fe2+/100 g seeds. The total phenolics content of grass pea extract was correlated with the results of the ABTS (r = 0.881) and FRAP (r = 0.781) assays. The same correlation was observed between the results of both assays (r = 0.842). Two derivatives of p-coumaric acid were the dominant phenolic compounds of the Derek cultivar of grass pea.

Phenolic contents and antioxidant capacities of wild and cultivated white lupin (Lupinus albus L.) seeds.

The aim of this study was to compare the antioxidant capacities and phenolic compound profiles of wild and cultivated Lupinus albus L. seeds. The total phenolic content (TPC), radical scavenging activity, ferric-reducing antioxidant power (FRAP) and antioxidant activity in an ß-carotene-linoleic acid emulsion were determined. Liquid chromatography-mass spectrometry was used to identify phenolics. The TPC of lupin seeds ranged from 4.36 to 7.24 mg gallic acid equivalent/g dry matter (d.m.). The dominant phenolics of all genotypes were two p-coumaric acid derivatives (0.74-1.61 and 0.66-1.63 mg/g d.m.) and apigenin-6,8-di-C-glucoside (1.13-1.31 mg/g d.m.). The results of antioxidant assays of wild lupin extracts were similar to or lower than those of the cultivated variety. FRAP and ABTS+ scavenging activity were correlated with the contents of the more polar p-coumaric acid derivative and apigenin-6,8-di-C-glucoside. Generally, significant differences between cultivated and wild L. albus seeds were not found in antioxidant capacities and phenolic compound contents.

The Structure-Antioxidant Activity Relationship of Ferulates.

The antioxidant activity of ferulic acid (1), iso-ferulic acid (2), coniferyl aldehyde (3), methyl ferulate (4), and ethyl ferulate (5) were investigated using 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric-reducing antioxidant power (FRAP) assays and autoxidation of triacylglycerols of commercially available sunflower oil (TGSO). The compounds tested for ability to scavenge ABTS radical cations was in the order of ferulic acid > coniferyl aldehyde ≈ iso-ferulic acid > ethyl ferulate ≈ methyl ferulate. The results of the FRAP assay for ferulic acid, iso-ferulic acid, and coniferyl aldehyde were similar to and higher than those of methyl ferulate and ethyl ferulate. In the lipid system, iso-ferulic acid showed weak antioxidant activity. The other ferulates exhibited much stronger, yet similar, activities.

Antioxidant Activity and Phenolic Composition of a Red Bean (Phasoelus vulgaris) Extract and its Fractions.

- The activities of the crude acetonic extract of red bean and its two fractions were determined using a 0-carotene-linoleate model system as well as the total antioxidant activity (TAA), the total phenolics content (TPC), the DPPH radical-scavenging activity, and the reducing power assays. Results from the in vitro assays showed the highest values when tannins (fraction II) were tested. Specifically, the TAA of the tannins fraction was 4.37 mmol Trolox eq./g fraction; whereas, the crude extract and fraction I were 0.481 and 0.093 µmol Trolox eqi/mg extract or fraction, respectively. The content of total phenolics in fraction II was the utmost (612 mg/g); the tannins content, assayed by the vanillin method and expressed as absorbance units at 500 nm per I g, was 938. RP-HPLC- PAD-MS profiling revealed the presence of 33 compounds: quercetin arabinoglucoside, quercetin rutinoside, quercetin, p-hydroxybenzoic acid, and kaempferol rutinoside were the most abundant phenolics in the extract.

Use of Different Proteases to Obtain Flaxseed Protein Hydrolysates with Antioxidant Activity.

The antioxidant activity of flaxseed protein hydrolysates obtained using five different enzymes was evaluated. Proteins were isolated from flaxseed cake and were separately treated with papain, trypsin, pancreatin, Alcalase and Flavourzyme. The degree of hydrolysis (DH) was determined as the percentage of cleaved peptide bonds using a spectrophotometric method with o-phthaldialdehyde. The distribution of the molecular weights (MW) of the hydrolysis products was profiled using Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) and size exclusion-high performance liquid chromatography (SE-HPLC) separations. The antioxidant activities of the protein isolate and hydrolysates were probed for their radical scavenging activity using 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS(•+)) and photochemiluminescence (PCL-ACL) assays, and for their ferric reducing antioxidant power (FRAP) and ability to bind Fe(2+). The hydrolysates were more effective as antioxidants than the protein isolate in all systems. The PCL-ACL values of the hydrolysates ranged from 7.2 to 35.7 µmol Trolox/g. Both the FRAP and ABTS(•+) scavenging activity differed among the hydrolysates to a lower extent, with the ranges of 0.20-0.24 mmol Fe(2+)/g and 0.17-0.22 mmol Trolox/g, respectively. The highest chelating activity (71.5%) was noted for the pancreatin hydrolysate. In general, the hydrolysates obtained using Alcalase and pancreatin had the highest antioxidant activity, even though their DH (15.4% and 29.3%, respectively) and the MW profiles of the peptides varied substantially. The O2(•-) scavenging activity and the ability to chelate Fe(2+) of the Flavourzyme hydrolysate were lower than those of the Alcalase and pancreatin hydrolysates. Papain was the least effective in releasing the peptides with antioxidant activity. The study showed that the type of enzyme used for flaxseed protein hydrolysis determines the antioxidant activity of the hydrolysates.

Antioxidant Activity of Flaxseed Extracts in Lipid Systems.

The aim of this work was to compare the antioxidant activity of the extract of flaxseed and its alkaline hydrolysate in two model systems: lipid autoxidation of triacylglycerols of sunflower oil (TGSO)-in a homogeneous lipid media and during ß-carotene-linoleate emulsion system. In addition, pure lignans were tested. The material was defatted with hexane and then phenolic compounds were extracted using dioxane-ethanol (50:50, v/v) mixture. Carbohydrates were removed from the crude extract using an Amberlite XAD-16 column chromatography. The content of total phenolic compounds in the crude extract and after alkaline hydrolysis was determined using a Folin-Ciocalteu's phenol reagent. Individual phenolic compounds were determined by nordihydroguaiaretic acid (RP-HPLC) method in gradient system. The alkaline hydrolysis increased the content of total phenolics in the extract approximately by 10%. In the extracts of flaxseed, phenolic compounds were present in the form of macromolecular complex. In the alkaline hydrolysate, secoisolariciresinol diglucoside (SDG) was found as the main phenolic compound. Small amounts of p-coumaric and ferulic acids were also determined. SDG and both extracts were not able to inhibit effectively lipid autoxidation. The kinetics of TGSO autoxidation at 80 °C in absence and in presence of the extract before hydrolysis (EBH) and after hydrolysis (EAH) was monitored and compared with known standard antioxidants. Ferulic acid (FA) and butylated hydroxyl toluene (BHT) showed much higher antioxidant efficiency and reactivity than that of both extracts. Secoisolariciresinol (SECO) showed a higher activity in both model systems than SDG. However, the activity of SECO was much lower than that of nordihydroquaiaretic acid (NDGA).

Analysis of phenolic compounds and antioxidant abilities of extracts from germinating Vitis californica seeds submitted to cold stress conditions and recovery after the stress.

The material for this study consisted of stratified seeds of Vitis californica submitted to germination under optimum conditions (+25 °C) or under chill stress (+10 °C), also followed by recovery. It has been determined that the germinating seeds contain considerable amounts of tannins, catechins as well as phenolic acids such as gallic, p-coumaric, caffeic and ferulic acids. Gallic acid appeared in the highest amount in the germinating seeds (from 42.40-204.00 µg/g of fresh weight (FW)), followed by caffeic acid (from 6.62-20.13 µg/g FW), p-coumaric acid (from 2.59-5.41 µg/g FW), and ferulic acid (from 0.56-0.92 µg/g FW). The phenolic acids occurred mostly in the ester form. Under chill stress, the germinating seeds were determined to contain an elevated total amount of phenolics, as well as raised levels of condensed tannins, catechins, gallic acid, and gafeic acid. The levels of p-coumoric and ferulic acids were found to have decreased. In extracts isolated from a sample exposed to low temperature, increased antioxidant activity and reduction potential were also demonstrated. Tissue of the germinating seeds which underwent post-stress recovery was found to have less total phenolics.

Differences in the phenolic composition and antioxidant properties between Vitis coignetiae and Vitis vinifera seeds extracts.

Phenolic compounds were extracted from European and Japanese grapevine species (Vitis vinifera and V. coignetiae) seeds using 80% methanol or 80% acetone. The total content of phenolic compounds was determined utilizing Folin-Ciocalteu's phenol reagent, while the content of tannins was assayed by the vanillin and BSA precipitation methods. Additionally, the DPPH free radical and ABTS cation radical scavenging activities and the reduction power of the extracts were measured. The HPLC method was applied to determine the phenolic compounds, such as phenolic acids and catechins. The seeds contained large amounts of tannins and gallic acid and observable quantities of catechins, p-coumaric, ferulic and caffeic acids. The dominant form of phenolic acids in the extracts was the ester-bound form. The content of total phenolics was higher in the European grape V. vinifera seeds, which also contained more tannins, catechins and phenolic acids, except for caffeic acid. Extracts from V. vinifera seeds showed better radical scavenger properties and stronger reducing power. The total contents of phenolic compounds and tannins in acetone extracts were higher than in methanolic extracts. Acetone extracts also exhibited stronger antiradical properties as well as stronger reducing power.

Extracts of phenolic compounds from seeds of three wild grapevines-comparison of their antioxidant activities and the content of phenolic compounds.

Phenolic compounds were extracted from three wild grapevine species: Vitis californica, V. riparia and V. amurensis seeds using 80% methanol or 80% acetone. The total content of phenolic compounds was determined utilizing the Folin-Ciocalteu's phenol reagent while the content of tannins was assayed with the vanillin and BSA precipitation methods. Additionally, the DPPH free radical scavenging activity and the reduction power of the extracts were measured. The RP-HPLC method was applied to identify the phenolic compounds in the extracts, such as phenolic acids and catechins. The seeds contained large amounts of tannins, catechins and gallic acid and observable quantities of p-coumaric acid. The total content of phenolic compounds and tannins was similar in the extracts from V. californica and V. riparia seeds. However, the total content of total phenolic compounds and tannins in the extracts from V. californica and V. riperia seeds were about two-fold higher than that in the extracts from V. amurensis seeds. Extracts from seeds of the American species (V. californica and V. riparia) contained similarly high concentrations of tannins, whereas extracts from seeds of V. amurensis had approximately half that amount of these compounds. The content of catechin and epicatechin was similar in all extracts. The highest DPPH(•) anti-radical scavenging activity was observed in the acetonic and methanolic extracts of V. californica and V. riparia seeds- while the acetonic extract from the V. californica seeds was the strongest reducing agent.

Phenolic compound profiles and antioxidant capacity of Persea americana Mill. peels and seeds of two varieties.

Avocado processing by the food and cosmetic industries yields a considerable amount of phenolic-rich byproduct such as peels and seeds. Utilization of these byproducts would be favorable from an economic point of view. Methanolic (80%) extracts obtained from lyophilized ground peels and seeds of avocado (Persea americana Mill.) of the Hass and Shepard varieties were characterized for their phenolic compound profiles using the HPLC-PAD technique. The structures of the identified compounds were subsequently unambiguously confirmed by ESI-MS. Compositional analysis revealed that the extracts contained four polyphenolic classes: flavanol monomers, proanthocyanidins, hydroxycinnamic acids, and flavonol glycosides. The presence of 3-O-caffeoylquinic acid, 3-O-p-coumaroylquinic acid, and procyanidin A trimers was identified in seeds of both varieties. Intervarietal differences were apparent in the phenolic compound profiles of peels. Peels of the Shepard variety were devoid of (+)-catechin and procyanidin dimers, which were present in the peels of the Hass variety. Peels of both varieties contained 5-O-caffeoylquinic acid and quercetin derivatives. The differences in the phenolic profiles between varietals were also apparent in the different antioxidant activity of the extracts. The peel extracts had a higher total phenolic compound content and antioxidant activity when compared to the seed extracts. The highest TEAC and ORAC values were apparent in peels of the Haas variety in which they amounted to 0.16 and 0.47 mmol Trolox/g DW, respectively. No significant (p > 0.05) differences were apparent between the TEAC values of seeds of the two varieties but the ORAC values differed significantly (p < 0.05). Overall these findings indicate that both the seeds and peel of avocado can be utilized as a functional food ingredient or as an antioxidant additive.

Limitations of the tetramethylmurexide assay for investigating the Fe(II) chelation activity of phenolic compounds.

Limitations of the colorimetric assay involving tetramethylmurexide (TMM) to determine the extent of complex formation between metal ions and phenolic compounds have been studied. Older literature reports using this method to determine bound Fe(II). Our study shows the TMM assay is inadequate when determining the Fe(II) chelation activity of phenolic preparations rich in tannin constituents on account of the high absorbance values derived by control samples (i.e., those that do not contain the TMM reagent). Phenolic test samples comprising the TMM reagent, iron ions, and tannins could not yield meaningful absorbance data on Fe(II) chelation activity. In our study, we investigated commercially available compounds, namely, sinapic acid, catechin, rutin, tannic acid, procyanidin B(2), as well as crude acetonic extracts of almonds, red lentil, buckwheat, and their low-molecular-weight and tannin fractions separated from the crude extracts by Sephadex LH-20 column chromatography. Even as little as 0.5 mg of tannins added per control sample resulted in high absorbance values to the extent of 0.4 for red lentil and almonds, and 1.3 for buckwheat. A strong correlation (r(2) = 0.98) between the content of condensed tannins, as determined by the vanillin reaction, and absorbance of control samples by the TMM assay was found for the plant extracts and their fractions. A more useful colorimetric assay to investigate the Fe(II) chelating ability of tannin-rich preparations may be the method that uses ferrozine.