Enzymatic preparation of kaempferol from green tea seed and its antioxidant activity

Among the flavonols in green tea, kaempferol has many biological activities but kaempferol of plant origin is too expensive to be used in commercial products. Recently, we confirmed that green tea seed (GTS) contained a reasonable amount of kaempferol glycoside. After conducting structure analysis, two kaempferol glycosides were identified, kaempferol-3-O-[2-O-beta-D-galactopyranosyl-6-O-alpha-L-rhamnopyranosyl]-beta-D-glucopyranoside (compound 1) and kaempferol-3-O-[2-O-beta-D-xylopyranosyl-6-O-alpha-L-rhamnopyranosyl]-beta-D-glucopyranoside (compound 2), respectively. Also, a commercially useful method for kaempferol preparation was suggested by enzymatic hydrolysis using these two flavonoids. After several enzyme reactions were performed for the complete bioconversion of compounds 1 and 2 to kaempferol, we found that the optimum enzyme combination was reaction with beta-galactosidase and hesperidinase. Finally, we produced pure kaempferol with over 95% purity. We also compared the antioxidant effect of these two GTS flavonoids and its aglycone, kaempferol. Kaempferol is a more efficient scavenger of 1,1-diphenyl-2-picrylhydrazyl radicals and a better inhibitor of xanthine/xanthine oxidase than the two glycosides.     

Enhancement of dissolution and antioxidant activity of kaempferol using a nanoparticle engineering process

Kaempferol (KAE) is a strong antioxidant flavonoid compound, but its clinical application is limited by quantity and poor dissolution property. However, the dissolution mechanism of a kaempferol nanoparticle formulation (KAEN) has not yet been elucidated. The aim of the present study was therefore to use a nanoparticle engineering process to resolve the dissolution problem. Our data indicated that KAEN effectively increased the dissolution percentage by particle size reduction, high encapsulation efficiency, amorphous transformation, and hydrogen-bond formation with excipients. In addition, we used several different antioxidant activity assays to evaluate KAE and KAEN. The data indicated that KAEN retained potent antioxidant activity after the nanoparticle engineering process and showed better antioxidant activity than KAE dissolved in water (P < 0.05). According to these findings, we concluded that KAEN could be a low-dose alternative to KAE in health food and future clinical research.     

Radiolysis of quercetin in methanol solution: observation of depside formation

Radiolysis of the flavonol quercetin, a natural antioxidant, was performed in methanol. The degradation process was followed by HPLC analyses. The major product was identified as a depside (Q1) by NMR and LC-MS. The G(Q1) radiolytic factor was plotted versus the initial concentration of quercetin. This radiolytic process was attributed to the CH3O* radicals presented in the irradiated medium. The proposed mechanism invoked a stereospecific oxidation of the 3-hydroxyl group of quercetin which led to C-ring opening and to the formation of the depside Q1. In presence of water, Q1 was transformed into another depside, Q2, by an inverse esterification reaction. A chemical equilibrium was observed between Q1 and Q2. The comprehension of the radiolytic process of quercetin in methanol solution is of importance. Indeed, the same type of oxidative reactions could occur on flavonoids during preservation of food by ionizing radiation.     

Mechanochemical-assisted extraction and antioxidant activities of kaempferol glycosides from Camellia oleifera Abel. meal

A mechanochemical-assisted extraction (MCAE) method was proposed and investigated for the fast extraction of two kaempferol glycosides (kaempferol-3-O-[2-O-β-D-galactopyranosyl-6-O-α-L-rhamnopyranosyl]-β-D-glucopyranoside and kaempferol-3-O-[2-O-β-D-xylopyranosyl-6-O-α-L-rhamnopyranosyl]-β-D-glucopyranoside) from Camellia oleifera Abel. meal. The effects of operating parameters in terms of NaOH content, grinding time, extraction time, and ratio of solution to solid were evaluated by means of response surface methodology (RSM). Under the optimal conditions with a ratio of material to NaOH of 20:1 (g/g), a milling time of 15 min, and a ratio of solution to solid of 20:1 (mL/g) for 60 min, the maximum extraction yields of the two kaempferol glycosides reached 13.34 and 13.83%, respectively. The antioxidant activity of kaempferol glycosides extract was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay and ferric thiocyanate (FTC) assay. Compared with the heat reflux extraction (HRE) method, the yield and the antioxidant activities of the extracts from MCAE with water as solvent were higher and stronger.     

Evaluation of radical products from beta-alanine/sugar mixtures by use of GC-MS with the galvinoxyl radical

We have quantified the radical production of Maillard systems with beta-alanine and different sugars by a method that uses the free radical scavenger capacity of galvinoxyl (2,6-di-tert-butyl-alpha-(3, 5-di-tert-butyl-4-oxo-2,5-cyclohexadiene-1-ylidene)-p-tolyloxy) and a GC-MS separation and quantification. We have compared the results with the spectrophotometrical conclusions and highlight the importance of the radical production in the Maillard systems. The method is precise (CV < 5%), accurate (>97%), sensitive (limit of detection 10 ng/mL galvinoxyl), and may be used for the detection and quantification of radical activity in the chemical and biological systems.     

Antimicrobial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey. Var. pectinatus (Lamiaceae)

The essential oil, obtained by using a Clevenger distillation apparatus, and water-soluble (polar) and water-insoluble (nonpolar) subfractions of the methanol extract of Thymus pectinatus Fisch. et Mey. var. pectinatus were assayed for their antimicrobial and antioxidant properties. No (or slight) antimicrobial activity was observed when the subfractions were tested, whereas the essential oil showed strong antimicrobial activity against all microorganisms tested. Antioxidant activities of the polar subfraction and the essential oil were evaluated using 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide radical scavenging, and lipid peroxidation assays. The essential oil, in particular, and the polar subfraction of the methanol extract showed antioxidant activity. The essential oil was analyzed by GC/MS, and 24 compounds, representing 99.6% of the essential oil, were identified: thymol, gamma-terpinene, p-cymene, carvacrol, and borneol were the main components. An antimicrobial activity test carried out with fractions of the essential oil showed that the activity was mainly observed in those fractions containing thymol, in particular, and carvacrol. The activity was, therefore, attributed to the presence of these compounds. Other constituents of the essential oil, such as borneol, gamma-terpinene, and p-cymene, could be also taken into account for their possible synergistic or antagonistic effects. On the other hand, thymol and carvacrol were individually found to possess weaker antioxidant activity than the crude oil itself, indicating that other constituents of the essential oil may contribute to the antioxidant activity observed. In conclusion, the results presented here show that T. pectinatus essential oil could be considered as a natural antimicrobial and antioxidant source.     

Botanical origin causes changes in nutritional profile and antioxidant activity of fermented products obtained from honey

Honey as rich source of enzymatic and nonenzymatic antioxidants serves as health-promoting nutrient in the human body. Here, we present the first time a comparative study of nutritional profiles (e.g., acidities, sugar, organic acid profile, total polyphenolic, flavonoid content) for different unifloral, multifloral honeys and their fermented products, in correlation with their antioxidant activity. Additionally, an optimized method for HPLC separation of organic acids from honey was established. The total phenolic content of honey samples varied widely among the honey types compared to fermented products. High amounts of total flavonoids were quantified in heather honey, followed by raspberry, multifloral, black locust, and linden honey. A positive correlation between the content of polyphenols, flavonoids, and antioxidant activity was observed in honey samples. After fermentation, the flavonoid content of dark honey fermented products decreased significantly. Black locust and linden honeys are more suitable for fermentation because the decrease in antioxidant substances is less pronounced.     

Evaluation of the stability and antioxidant activity of nanoencapsulated resveratrol during in vitro digestion

Resveratrol was encapsulated in oil-in-water food-grade nanoemulsions of subcellular size, produced by high-pressure homogenization. Physicochemical stability was evaluated under accelerated aging (high temperature and UV light exposure), as well as during simulated gastrointestinal digestion. Antioxidant activity was assessed at different stages of digestion by chemical assays and by an improved cellular assay, to measure exclusively the residual activity of resveratrol that penetrated inside Caco-2 cells. Results showed that the nanoemulsions based on soy lecithin/sugar esters and Tween 20/glycerol monooleate were the most physically and chemically stable, in terms of mean droplet size (always <180 nm) and resveratrol loading, during both accelerated aging and gastrointestinal digestion. These formulations also exhibited the highest chemical and cellular antioxidant activities, which was comparable to unencapsulated resveratrol dissolved in DMSO, suggesting that nanoencapsulated resveratrol, not being metabolized in the gastrointestinal tract, can be potentially absorbed through the intestinal wall in active form.     

Evaluation of antioxidant activity of vetiver (Vetiveria zizanioides L.) oil and identification of its antioxidant constituents

Antioxidant capacities of vetiver (Vetiveria zizanioides) oil were evaluated by two different in vitro assays: the DPPH* free radical scavenging assay and the Fe2+-metal chelating assay. Results showed that the vetiver oil (VO) possessed a strong free radical scavenging activity when compared to standard antioxidants such as butylated hydroxytoluene (BHT) and alpha-tocopherol. However, its metal chelating capacity was relatively weak. VO (10 microL/mL) dissolved in methanol exhibited approximately 93% free radical scavenging activity in the DPPH* assay and approximately 34% Fe2+ chelating activity in the metal chelating assay. By contrast, 10 mM BHT and 0.1 mM alpha-tocopherol exhibited 93 and 89% free radical scavenging activities in the DPPH* assay, respectively, and 1 mM EDTA exhibited approximately 97% activity in the metal chelating assay. Among the complex constituents in the crude VO, beta-vetivenene, beta-vetivone, and alpha-vetivone, which had shown strong antioxidant activities, were isolated and identified using various chromatographic techniques including silica gel open column chromatography, silica HPLC, and GC-MS. These results show that VO and some of its inherent components can be potential alternative natural antioxidants.     

Screening methods to measure antioxidant activity of sorghum (sorghum bicolor) and sorghum products

Specialty sorghums, their brans, and baked and extruded products were analyzed for antioxidant activity using three methods: oxygen radical absorbance capacity (ORAC), 2,2'-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), and 2,2-diphenyl-1-picrylhydrazyl (DPPH). All sorghum samples were also analyzed for phenolic contents. Both ABTS and DPPH correlated highly with ORAC (R(2) = 0.99 and 0.97, respectively, n = 18). Phenol contents of the sorghums correlated highly with their antioxidant activity measured by the three methods (R(2) >or= 0.96). The ABTS and DPPH methods, which are more cost effective and simpler, were demonstrated to have similar predictive power as ORAC on sorghum antioxidant activity. There is a need to standardize these methods to allow for data comparisons across laboratories.     

A new test method for the evaluation of total antioxidant activity of herbal products

A new test method for measuring the antioxidant power of herbal products, based on solid-phase spectrophotometry using tetrabenzo-[b,f,j,n][1,5,9,13]-tetraazacyclohexadecine-Cu(II) complex immobilized on silica gel, is proposed. The absorbance of the modified sorbent (lambda(max) = 712 nm) increases proportionally to the total antioxidant activity of the sample solution. The method represents an attractive alternative to the mostly used radical scavenging capacity assays, because they generally require complex long-lasting stages to be carried out. The proposed test method is simple ("drop and measure" procedure is applied), rapid (10 min/sample), requires only the monitoring of time and absorbance, and provides good statistical parameters (s(r)相似文献   

Study of the polyphenolic composition and antioxidant activity of new sherry vinegar-derived products by maceration with fruits

Several experiments of maceration of a sherry wine vinegar with different fruits (orange, lemon, strawberry, grapefruit, and lime) have been carried out. After optimization (only peel, no heating and seven days as maximum time of maceration), parameters such as polyphenolic content, superoxide anion scavenging ability (related to antioxidant activity) and ascorbic acid content were determined in sherry wine vinegars macerated with two amounts of peel and for two maceration times (3 and 7 days). The analysis of variance pointed to a clear relationship (p<0.01) between type of fruit and amount of peel and polyphenolic content. The factor "time" was practically not significant for any polyphenol. Sherry wine vinegars macerated with different fruits exhibited higher superoxide anion scavenger ability, with the maximum values found for the vinegar macerated with lemon peel. The correlation analysis showed that the superoxide anion scavenger ability of the vinegars macerated, and thus their antioxidant activity, was highly correlated (p<0.01) with several polyphenols, especially with naringin, hesperidin, neohesperidin and gentisic acid and not with the ascorbic acid content.     

Evaluation of the phytoestrogenic activity of Cyclopia genistoides (honeybush) methanol extracts and relevant polyphenols

Unfermented C. genistoides methanol extracts of different harvestings and selected polyphenols were evaluated for phytoestrogenic activity by comparing binding to both ER subtypes, transactivation of an ERE-containing promoter reporter, proliferation of MCF-7-BUS and MDA-MB-231 breast cancer cells, and binding to SHBG. The extracts from one harvesting of C. genistoides (P104) bound to both ER subtypes. All extracts transactivated ERE-containing promoter reporters via ERbeta but not via ERalpha. All extracts, except P122, caused proliferation of the estrogen-sensitive MCF-7-BUS cells. Proliferation of MCF-7-BUS cells was ER-dependent as ICI 182,780 reversed proliferation. Physiologically more relevant, extracts antagonized E2-induced MCF-7-BUS cell proliferation. Furthermore, all extracts, except P122, induced proliferation of the estrogen-insensitive MDA-MB-231 cells, suggesting that the extracts are able to induce ER-dependent and ER-independent cell proliferation. Binding to SHBG by extracts was also demonstrated. These results clearly show that C. genistoides methanol extracts display phytoestrogenic activity and act predominantly via ERbeta. HPLC and LC-MS analysis, however, suggests that the observed phytoestrogenic activity cannot be ascribed to polyphenols known to be present in other Cyclopia species.     

Evaluation of the antioxidant activity of capsiate analogues in polar, nonpolar, and micellar media

Synthesis of 10 capsiate analogues was conducted by lipase-mediated (Novozyme 435) esterification of vanillyl alcohol with different fatty acids. The antioxidant activity of the synthesized capsiates was evaluated using three in vitro assays: DPPH radical scavenging assay (polar medium), Rancimat assay (nonpolar medium), and autoxidation of linoleic acid (micellar medium). The objective of this study is to find the influence of structural characteristics of the alkyl chain of capsiate analogues on their antioxidant activity. In these assays, BHT and α-tocopherol were used as reference compounds. Both DPPH and Rancimat assays did not show any specific trend of antioxidant activity with the increase in lipophilicity and also with the type of fatty acids grafted to the phenolic moiety. In the Tween 20 micellar system for the inhibition of autoxidation of linoleic acid, vanillyl ester attached to a C18 alkyl chain (vanillyl stearate, oleate, and ricinoleate) exhibited maximum inhibition of autoxidation of linoleic acid.     

Method for measuring antioxidant activity and its application to monitoring the antioxidant capacity of wines.

A novel method for measuring the antioxidant activity using N, N-dimethyl-p-phenylenediamine (DMPD) was developed. The radical cation of this compound gives a stable colored solution and a linear inhibition of color formation can be observed in the presence of 0. 2-11 microg of TROLOX. The experimental protocol, which is rapid and inexpensive, ensures sensitivity and reproducibility in the measure of antioxidant activity of hydrophilic compounds. The effectiveness of the DMPD method on real foods was verified by evaluating the antioxidant ability of wine samples coming from different areas of Campania, Italy. Antioxidant capacity of wines is strictly related to the amount of phenolic compounds. The results obtained by the DMPD method are very similar to those obtained on the same samples when the radical cation of 2, 2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (Miller et al., 1996) was used.     

Evaluation of antioxidant activity and preventing DNA damage effect of pomegranate extracts by chemiluminescence method

The antioxidant activities of three parts (peel, juice, and seed) and extracts of three pomegranate varieties in China were investigated by using a chemiluminescence (CL) method in vitro. The scavenging ability of pomegranate extracts (PEs) on superoxide anion, hydroxide radical, and hydrogen peroxide was determined by the pyrogallol-luminol system, the CuSO4-Phen-Vc-H2O2 system, and the luminol-H2O2 system, respectively. DNA damage preventing the effect of PE was determined by the CuSO4-Phen-Vc-H2O2-DNA CL system. The results showed that the peel extract of red pomegranate had the best effect on the scavenging ability of superoxide anion because its IC50 value (4.01 +/- 0.09 microg/mL) was the lowest in all PEs. The seed extract of white pomegranate could scavenge hydroxide radical most effectively of the nine extracts (the IC50 value was 1.69 +/- 0.03 microg/mL). The peel extract of white pomegranate had the best scavenging ability on hydrogen peroxide, which had the lowest IC50 value (0.032 +/- 0.003 microg/mL) in the nine extracts. The seed extract of white pomegranate (the IC50 value was 3.67 +/- 0.03 microg/mL) was the most powerful on the DNA damage-preventing effect in all of the PEs. Also, the statistical analysis indicated that there were significant differences (at P < 0.05) among the extracts of the different varieties and parts in each system.     

Evaluation of the antioxidant activity of environmental plants: activity of the leaf extracts from seashore plants

The antioxidant activity of the methanolic extracts of the leaves of 39 plant species was examined. These leaves were collected from the plants growing on subtropical seashores. The activity was evaluated by three kinds of assay methods, which included the DPPH radical scavenging assay, linoleic acid oxidation assay, and oxidative cell death assay. Two extracts from Excoecaria agallocha and Terminalia catappa showed remarkably potent activity in all assay systems. The HPLC analysis of the extracts indicated the presence of the same antioxidant and isolation work for the compound identified ellagic acid. The isolated ellagic acid showed strong antioxidant activity in the assay systems used.     

Evaluation of antioxidant activity and inhibitory effect on nitric oxide production of some common vegetables

The objectives of this study were to study the antioxidant activities and nitric oxide (NO) scavenging effects of vegetables in vitro systems and to study the inhibitory effects of vegetables on the NO production and NO-induced DNA damage in RAW 264.7 macrophage. The results indicated that water extracts from Indian lotus, Jew's ear, shiitake, eggplant, and winter mushroom showed stronger antioxidant activity and free-radical-scavenging ability than that of other vegetable extracts. The scavenging effects of vegetable extracts on NO derived from sodium nitroprusside (SNP) were in decreasing order of water spinach > Indian lotus > eggplant and garland chrysanthemum. In the macrophage model system, the water extracts from fresh daylily flower, sponge gourd, pea sprout, and eggplant exhibited over 80% inhibition on NO generation stimulated by lipopolysaccharide. The extract from fresh daylily flower that expressed the strongest inhibition on NO production was attributed to the ability to reduce the inducible nitric oxide synthase (iNOS) induction. However, the extracts from pea sprout and eggplant suppressed the NO production by scavenging on NO and inactivating toward iNOS enzyme. In addition, the water extracts from fresh daylily flower, sponge gourd, pea sprout, and eggplant also showed over 40% inhibitory effect on DNA damage induced by SNP in RAW 264.7 macrophage. The data also indicated that eggplant and pea sprout extracts contained higher total phenolic compounds, anthocyanins, and ascorbic acids and appeared to be responsible for their antioxidant activities and scavenging effects on NO derived from SNP.     

Evaluation of antioxidant activity and electronic taste and aroma properties of antho-beers from purple wheat grain

Moderate consumption of beer is known to be beneficial for health. Thus, antioxidant, likely taste, and aroma properties of antho-beers made from purple wheat grain (antho-grain) were evaluated. The 2,2-diphenyl-1-picryhydrazyl free radical (DPPH*) scavenging activity, total phenolic content (TPC), oxygen radical absorbance capacity (ORAC), and phenolic acid compositions of antho-bran were also investigated. DPPH* scavenging activity at 60 min was 50.6-59.9% for control and antho-beer extracts, 15.0-54.1% for antho-bran extracts and hydrolysates. The TPC ranged from 410 to 609 mg/L, from 84 to 95 mg/L, and from 2473 to 7634 mg/kg for control (from barley malt) and antho-beer original samples, control and antho-beer extracts, and antho-bran extracts and hydrolysates, respectively. The corresponding ORAC values were 3050-4181 mg/L, 2961-3184 mg/L, and 74-213 g/kg, respectively. The major known phenolic acids comprised four types in control beer, five types in antho-beers, and seven types in antho-bran hydrolysates. Total anthocyanin content of antho-bran was up to 1160 mg/kg. Differences in likely taste and aroma were found between control and antho-beers by using electronic tongue and nose methods. Brewing materials had an effect on the antioxidant, likely taste, and aroma properties of beers; however, antho-grain may have potential as a novel brewing material.