Antioxidant and tyrosinase inhibitory activities of different parts of guava (<Emphasis Type="Italic">Psidium guajava</Emphasis> L.)

The antioxidant and the tyrosinase inhibitory activities of 4 different solvents (acetone, ethanol, methanol, and water) for preparation of extracts from guava (branch, fruit, leaf, and seed) were evaluated by measuring total phenolic contents (TPC), DPPH radical scavenging activity, ABTS radical scavenging activity, reducing power (RP), and tyrosinase inhibitory activity. The extracts of branch and leaf showed relatively higher antioxidant properties than those of fruit and seed. The highest TPC (141.28 mg/g gallic acid equivalents), DPPH radical scavenging activity (IC₅₀=34.01 μg/mL), ABTS radical scavenging activity (IC₅₀=3.23 μg/mL), and RP (IC₅₀= 75.63 μg/mL) were found in acetone extract of leaf, while water extract of seed had the lowest antioxidant activity. The tyrosinase inhibitory activity of ethanol extract from guava leaf was 69.56%, which was the highest activity among the extracts. These results indicate that useful bioactive substances exist in the guava branch as well as leaf extracts.     

Antioxidative and ACE inhibitory activities in enzymatic hydrolysates of the cotton leafworm, Spodoptera littoralis

The larvae of the cotton leafworm, Spodoptera littoralis, were used as a source of food proteins exerting possible biological activities. A simulated gastrointestinal digestion (IC₅₀ = 320 μg/ml) and digestion by mucosal enzymes (IC₅₀ = 211 μg/ml) reveals a significantly higher in vitro ACE inhibitory activity compared to hydrolysis using thermolysin (IC₅₀ = 1392 μg/ml) and alcalase (IC₅₀ = 827 μg/ml) as pretreatment. This indicates that the choice of enzymes to generate ACE inhibitory peptides is important. All hydrolysates were also tested for antioxidant activity using two tests: a radical scavenging test using DPPH and the ferric reducing antioxidant power (FRAP) assay, and they showed a similar antioxidant activity which was relatively low compared to the standard antioxidants BHT and vitamin C. As a conclusion, the data obtained suggest that insect protein can be used to generate hydrolysates, exerting both ACE inhibitory and antioxidant activity, which might be incorporated as multifunctional ingredient into functional foods.     

Comparative evaluation of rosmarinic acid,methyl rosmarinate and pedalitin isolated from Rabdosia serra (MAXIM.) HARA as inhibitors of tyrosinase and α-glucosidase

Rabdosia serra has been used in traditional Chinese medicine for centuries. In order to illustrate the pharmaceutical activity of R. serra as hypoglycaemic and skin-whitening agents, rosmarinic acid (confirmed as the major compound in R. serra), methyl rosmarinate and pedalitin isolated from R. serra were evaluated for their inhibitory effects and mechanisms on tyrosinase and α-glucosidase. The inhibitory effects on both tyrosinase and α-glucosidase were in decreasing order, pedalitin > methyl rosmarinate > rosmarinic acid. The IC₅₀ values for the tyrosinase and α-glucosidase activity inhibited by pedalitin were 0.28 and 0.29 mM, respectively. Both rosmarinic acid and methyl rosmarinate were considered as noncompetitive inhibitors of tyrosinase, while pedalitin was suggested to be a mixed-type inhibitor of tyrosinase. In the assay of α-glucosidase inhibition, rosmarinic acid was found to be a competitive inhibitor, whereas both methyl rosmarinate and pedalitin were considered as mixed-type inhibitors.     

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

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

Antioxidant and free radical scavenging activities of wild bitter melon (Momordica charantia Linn. var. abbreviata Ser.) in Taiwan

Momordica charantia Linn. var. abbreviata Ser. (Cucurbitaceae), also known as “Shan Ku Gua”, is a wild variety of bitter melon (BM) in Taiwan. The size of its fruits is only about one-fifth of the commonly seen BM. It is commonly consumed as vegetable and also used as a popular folk medicine. In this study, the antioxidant and free radical scavenging activities of BM aqueous (BM-H₂O) and ethanol (BM-EtOH) extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), metal chelation, cytochrome c and xanthine oxidase inhibition (XOI) assays, as well as FeCl₂-ascorbic acid induced lipid peroxidation (thiobarbituric acid reactive substances, TBARS) assay in rat liver homogenates in vitro. Total flavonoid and phenol contents of BM extracts were also analyzed. Results showed that both BM-H₂O (IC₅₀=129.94 μg/ml) and BM-EtOH (IC₅₀=156.78 μg/ml) possess potent DPPH radical scavenging activity, which was better than vitamin E (IC₅₀=172.21 μg/ml). These extracts also showed better iron chelating activity than vitamin E. However, they were weaker than vitamin E in free radical scavenging, xanthine oxidase inhibitory and anti-lipid peroxidation activities. With the exception of XOI activity [IC₅₀=7.90 μg/ml (BM-H₂O) vs. 7.69 μg/ml (BM-EtOH)], BM-H₂O showed a lower IC₅₀ value in free radical scavenging [IC₅₀=6.15 μg/ml (BM-H₂O) vs. 7.08 μg/ml (BM-EtOH)] and anti-lipid peroxidation [IC₅₀=53.72 μg/ml (BM-H₂O) vs. 88.51 μg/ml (BM-EtOH) for liver; 82.53 μg/ml (BM-H₂O) vs. 91.83 μg/ml (BM-EtOH) for brain] activities than BM-EtOH. Both BM extracts showed a weak anti-lipid peroxidation activity in plasma. BM-H₂O (62.0 mg/g) possessed a significant higher concentration of total flavonoids than BM-EtOH (44.0 mg/g), but was lower in the total phenol content (BM-H₂O: 51.6 mg/g vs. BM-EtOH: 68.8 mg/g). In conclusion, BM extracts possess potent antioxidant and free radical scavenging activities. These antioxidant activities could have contributed, at least partly, to the therapeutic benefits of the certain traditional claims of wild BM.     

Evaluation of antioxidant activity of aqueous extract of some selected nutraceutical herbs

The objectives of this study were to examine the antioxidant activities and free radical scavenging effects of extracts of aqueous leaves of Psidium guajava L. (PE), Camellia sinensis (GABA tea; CE), Toona sinensis Roem. (TE) and Rosemarinus officinalis L. (RE). Among the four extracts, PE exhibited the strongest efficiency and showed over 50% scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at the concentration of 100 μg/mL. The reducing power of four nutraceutical herbs was in the order of PE > RE > CE > TE. The antioxidant activities of nutraceutical herbs were evaluated in a liposomes oxidation system promoted by Fe³⁺/ascorbic acid/H₂O₂. PE still showed the strongest antioxidant activity and exhibited over 95% inhibition at concentration of 50 μg/mL. The antioxidant activity of TE was still lower than that of other herbal plants; however, it also displayed 89% inhibition at concentration of 250 μg/mL. RE exhibited well inhibitory effects on the UVB-induced oxidation of erythrocyte ghosts at lower concentration (100 μg/mL). However, the protection of PE on the UVB-induced oxidation was significantly raised with increasing the concentrations and reached 95.4% inhibitory effects at concentration of 500 μg/mL. These results show that the tested herbal tea, especially PE could be considered as a natural antioxidant source.     

Sweet potato (Ipomoea batatas [L.] Lam ‘Tainong 57’) storage root mucilage with antioxidant activities in vitro

Sweet potato storage root mucilage was extracted and purified by SDS and heating treatments. Total antioxidant activity, DPPH (1,1-diphenyl-2-picrylhydrazyl) staining, reducing power method, metal ion-dependent hydroxyl radical, FTC (ferric thiocyanate) method, and protection of calf thymus DNA against hydroxyl radical-induced damage were studied. Half-inhibition concentrations, IC₅₀, were 0.08 mg/ml and IC₅₀ > 0.1 mg/ml, respectively, for the crude and purified mucilage in the total antioxidant activity test. In the DPPH staining, the crude and purified mucilage appeared as white spots when they were diluted to 50 and 100 μg per application, respectively. Like total antioxidant activity, reducing power, scavenging capacity against hydroxyl radical, FTC activity and protection of calf thymus DNA against hydroxyl radical-induced damage were found in the mucilage. It is suggested that the mucilage might contribute its antioxidant activities against both hydroxyl and peroxyl radicals.     

Dioscin: A synergistic tyrosinase inhibitor from the roots of Smilax china

In our study, we investigated the inhibition of mushroom tyrosinase in Smilax china. A methanol (MeOH) extract of S. china was partitioned into hexane, ethyl acetate (EtOAc) and water. Of the three fractions, EtOAc extract showed the strongest inhibition of tyrosinase activity with l-tyrosine or l-DOPA as a substrate. Two compounds were isolated from a final active fraction by activity-guided column chromatography. These compounds were identified as dioscin and oxyresveratrol by comparing their mass, ¹H- and ¹³C-NMR spectral data with those reported in the literature. Dioscin showed little inhibition activity of tyrosinase, whereas oxyresveratrol, a known tyrosinase inhibitor, showed a strong tyrosinase inhibitory activity. We discovered that a mixture of oxyresveratrol and dioscin (IC₅₀ = 5.1 and 5.7 μg/ml) highly increased the inhibition of tyrosinase activity with l-tyrosine or l-DOPA as the substrate as compared to either oxyresveratrol (IC₅₀ = 7.8 and 10.9 μg/ml) or dioscin (IC₅₀ > 100 and 100 μg/ml) alone.     

Inhibitory effect of raspberries on starch digestive enzyme and their antioxidant properties and phenolic composition

Seven primocane fall-bearing raspberry (Rubus idaeus L.) cultivars, Nova (red), Dinkum (red), Heritage (red), Autumn Britten (red), Josephine, Anne (yellow), Fall Gold (yellow) were analysed for potential health promoting properties including their inhibitory effect on starch and fat digestive enzymes, antioxidant activities, and phenolic composition. The tested raspberry extracts showed no detectable inhibition of pancreatic α-amylase and lipase. However, all the extracts exhibited potent inhibition of α-glucosidase with IC₅₀ from 16.8 to 34.2 μg/mL. Four phenolic compounds, ellagic acid, cyanidin-diglucoside, pelargonidin-3-rutinoside, and catechin were identified as the active α-glucosidase inhibitors. The raspberry extracts also possessed significant antioxidant activities with oxygen radical absorbance capacities (ORAC) ranging from 136.7 to 205.2 μmol Trolox equivalents (TE)/g dry weight fruit and DPPH radical scavenging activities from 305 to 351 μmol TE/g. The total phenolic content of raspberry cultivars varied significantly from 40.9 to 98.5 mg of gallic acid equivalents/g dry weight. The anthocyanin content varied widely from 0.1 to 9.5 mg cyanidin 3-glucoside equivalents/g. Nine phenolic acids were quantified in raspberries and their total amounts varied from 157.3 to 713.5 μg/g. The enzyme inhibition and antioxidant properties of raspberry cultivars were not correlated with their total phenolic, anthocyanin, and phenolic acid content. Overall, ‘Dinkum’ and ‘Josephine’ raspberry varieties possess higher total phenolic content, ORAC, DPPH radical scavenging activity, and α-glucosidase inhibitory activity than other five cultivars.     

Synthesis and structure–activity relationships and effects of phenylpropanoid amides of octopamine and dopamine on tyrosinase inhibition and antioxidation

Phenylpropanoid amides of octopamine (OA) 1a–1e and dopamine (DA) 2a–2e were synthesised and the structure–activity relationships (SARs) for antioxidant and tyrosinase inhibition activities were analysed. Among synthesised compounds, 2c, which contains two catechol moieties, exhibited the most DPPH radical-scavenging activity (EC₅₀ = 16.2 ± 2.4 μM), and 1d exhibited significant tyrosinase inhibitory activity (IC₅₀ = 5.3 ± 1.8 μM). Interestingly, with the same acid moiety, OA derivatives showed more inhibitory effect on tyrosinase than did compounds derived from DA, whereas DA derivatives were found to have higher antioxidant activity than compounds derived from OA. The relationship between their structures and their potencies, demonstrated in the current study, will be useful for the design of optimal agents.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Antioxidant and free radical scavenging activities of pigments extracted from molasses alcohol wastewater

The pigments from molasses alcohol wastewater were extracted by the macroporous resin adsorption method. The antioxidant and free radical scavenging activities of these pigments were also investigated. The adsorptive characteristics of five macroporous resins including HPD-600, HPD-500, D301-R, NKA-II and D296-R were studied and the results showed that the macroporous resin HPD-600 was most appropriate for extracting the pigments from molasses alcohol wastewater. The antioxidant and free radical scavenging activities of pigments extracted from alcohol wastewater were evaluated using nitrate, hydroxyl radical, superoxide anion radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) in vitro model systems. The pigment extract exhibited a concentration-dependent radical scavenging activity in all the systems. Meanwhile, scavenging activity of pigment extract in the DPPH system was found to be significantly (P < 0.05) higher than that in other systems and the 50% inhibitory concentration (IC₅₀ value) was about 0.07 mg/ml. The scavenging effect of pigment extract on superoxide anion radical was very weak with IC₅₀ value greater than 10 mg/ml.     

Inhibitory effects of α-cyano-4-hydroxycinnamic acid on the activity of mushroom tyrosinase

The effects of α-cyano-4-hydroxycinnamic acid (HCCA) on the activity of mushroom tyrosinase have been studied. Results showed that HCCA could inhibit both the monophenolase activity and diphenolase activity of mushroom tyrosinase. For the monophenolase activity, the lag phase was obviously lengthened, and the steady-state activity of the enzyme decreased sharply. When the concentration of HCCA reached to 80 μM, the lag time was lengthened from 20 s to 150 s and the steady-state activity was lost by about 75%. The IC₅₀ value was estimated to be 48 μM. For the diphenolase activity, the inhibitory effect of HCCA was also dose-dependent and the IC₅₀ value was estimated to be 2.17 mM. The kinetic analyses showed that the inhibition of HCCA on the diphenolase activity was reversible and competitive with the inhibition constants (K_I) determined to be 1.24 mM.     

Tyrosinase inhibitory effects and antioxidative activities of novel cinnamoyl amides with amino acid ester moiety

Nine cinnamoyl amides with amino acid ester (CAAE) moiety were synthesized by the conjugation of the corresponding cinnamic acids (cinnamic acid, 4-hydroxy cinnamic acid, ferulic acid and caffeic acid) with amino acid esters, and their inhibitory effects on the activities of mushroom tyrosinase were investigated, using l-3,4-dihydroxyl-phenylalanine (l-DOPA) as the substrate. Among these CAAE amides, ethyl N-[3-(4-hydroxy-3-methoxyphenyl)-1-oxo-2-propen-1-yl]-l-phenylalaninate (b₄) showed the strongest inhibitory activity; the IC₅₀ was 0.18 μM. The IC₅₀ values, inhibition types, inhibition mechanisms and kinetics of all these CAAE amides were evaluated. A structure–activity relationship (SAR) study found that the inhibitory effects were potentiated with the increasing length of hydrocarbon chains at the amino acid esters and also influenced by the substituents at the styrene groups. Furthermore, the hydroxyl radical scavenging and anti-lipid peroxidation activities of four CAAE derivatives were also investigated. Among these compounds, b₃ (ethyl N-[3-(3,4-dihydroxyphenyl)-1-oxo-2-propen-1-yl]-l-phenylalaninate) and b₄ exhibited potential antioxidant activities.     

DPPH antioxidant assay revisited

Scavenging of DPPH free radical is the basis of a common antioxidant assay. A number of protocols have been followed for this assay resulting in variation in the results of different laboratories. We present a perspective of the protocols followed by different workers with incongruity in their results and recommend a standard procedure within the sensitivity range of spectrophotometry. Three common standard antioxidants viz. ascorbic acid, BHT and propyl gallate have been used in this study. The IC₅₀ values for ascorbic acid and propyl gallate were 11.8 μM and 4.4 μM in methanol and 11.5 μM and 4.7 μM in buffered methanol as reaction medium, respectively. The free radical scavenging by BHT was markedly influenced by the reaction medium. The IC₅₀ values were 60.0 μM and 9.7 μM when the reaction was done in methanol and buffered methanol, respectively.     

Antioxidant and nitric oxide production inhibitory activities of galacturonyl hydroxamic acid

The self-prepared pectin hydroxamic acid has been reported to have antioxidant activities [Yang, S. S., Cheng, K. D., Lin, Y. S., Liu, Y. W., & Hou, W. C. (2004). Pectin hydroxamic acids exhibit antioxidant activities in vitro. Journal of Agricultural and Food Chemistry, 52, 4270–4273]. In this study, the galacturonic acid (GalA), the monomer unit of the pectin polymer, was esterified with acidic methanol (1 N HCl) at 4 °C with gentle stirring for 5 days to get galacturonic acid methyl ester which was further reacted with alkaline hydroxylamine to get galacturonyl hydroxamic acid (GalA–NHOH). The GalA–NHOH was used to test the antioxidant and antiradical activities in the comparison with GalA. The scavenging activities of GalA–NHOH against DPPH radicals (half-inhibition concentration, IC₅₀, was 82 μM), hydroxyl radicals detected by electron spin resonance (IC₅₀ was 0.227 nM in the comparison with Trolox of 0.433 μM), superoxide radicals (IC₅₀ was 830 μM) were determined. The protection activities of GalA–NHOH against hydroxyl radicals-mediated calf thymus DNA damages, linoleic acid peroxidation and peroxynitrite-mediated dihydrorhodamine 123 oxidations were also investigated. It was found that the GalA–NHOH exhibited dose-dependently antioxidant activity and few or none was found in GalA. The GalA–NHOH was used to evaluate the suppressed activity of nitric oxide (NO) productions of RAW264.7 cells in the presence of lipopolysaccharide (LPS, 100 ng/ml) as inducers. It was found that GalA–NHOH (0.02–0.1 mg/ml) could dose-dependently suppress the NO productions (expressed as nitrite concentrations) in RAW264.7 cells without significant cytotoxicity.     

The influence of fruit ripening on the phytochemical content and biological activity of Capsicum chinense Jacq. cv Habanero

During the past decade, it has been reported that the consumption of certain foods and spices such as pepper may have a positive effect on health. The present study evaluates the influence of fruit ripening on total phenols, flavonoids, carotenoids and capsaicinoids content and antioxidant, hypoglycaemic and anticholinesterase activities of Capsicum chinense Jacq. cv Habanero. The chemical investigation showed a different composition between the two stages of ripening (immature and mature). Generally, the concentration of carotenoids and capsaicinoids increased as the peppers reached maturity, whereas the concentration of phenols declined. The immature fruits showed the highest radical scavenging activity (IC₅₀ of 97.14 μg/ml). On the contrary, the antioxidant activity evaluated by the β-carotene bleaching test showed a significant activity for mature peppers (IC₅₀ value of 4.57 μg/ml after 30 min of incubation). Mature peppers inhibited α-amylase with an IC₅₀ of 130.67 μg/ml. The lipophilic fractions of both mature and immature peppers exhibited an interesting and selective inhibitory activity against α-amylase with IC₅₀ values of 29.58 and 9.88 μg/ml, respectively. Both total extracts of mature and immature peppers inhibited butyrylcholinesterase selectively. The obtained results underline the potential health benefits as a result of consuming C. chinense Habanero and suggest that it could be used as new valuable flavour with functional properties for food or nutriceutical products on the basis of the high content of phytochemicals and found biological properties.     

Inhibitory potential of trilobatin from Lithocarpus polystachyus Rehd against α-glucosidase and α-amylase linked to type 2 diabetes

The chemical structure of the sweet compound from Lithocarpuspolystachyus Rehd was identified as trilobatin on the basis of HPLC, EIS-MS and NMR analyses. The inhibitory activities of trilobatin against α-glucosidase and α-amylase were evaluated, and the inhibition mechanism was analysed with Lineweaver–Burk plots. Also the antioxidant activity evaluation of trilobatin was conducted by DPPH radical scavenging assay. Comparing with acarbose, trilobatin showed a strong inhibitory activity against α-glucosidase and a moderate inhibitory activity against α-amylase. The Lineweaver–Burk plots analysis elucidated that trilobatin inhibited the enzyme non-competitively. DPPH scavenging activity of trilobatin (IC₅₀ = 0.57 mg/ml) was higher than rutin (IC₅₀ = 0.72 mg/ml), which indicated that trilobatin had a moderate antioxidant potential. These results suggest that trilobatin is a potential effective α-glucosidase inhibitor for management of postprandial hyperglycemia with less side effect, and provide strong rationale for further animal and clinical studies.     

Antioxidant activity and proline content of leaf extracts from Dorystoechas hastata

Dorystoechas hastata (D. hastata) is a monotypic plant endemic to Antalya province of Turkey. D. hastata leaves are used to make a tea locally called “çalba tea”. Diethyl ether (E), ethanol (A), and water (W) were used for the sequential preparation of extracts from dried D. hastata leaves. A hot water extract (S) was also prepared by directly boiling the powdered plant in water. The antioxidant activities of the extracts were tested by ferric thiocyanate (FTC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging methods. E extract exhibited the greatest antioxidant activity with FTC method, whereas S extract exhibited the lowest IC₅₀ value (6.17 ± 0.53 μg/ml) for DPPH radical scavenging activity. Total phenolic contents of the extracts were estimated by Folin–Ciocalteu method and S extract was found to contain the highest amount (554.17 ± 20.83 mg GAE/g extract) of phenolics. Extract A contained highest flavonoid content and there was a inverse linear correlation (R² = 0.926) between IC₅₀ values for DPPH radical scavenging activity and flavonoid contents of all extracts. Reducing power of extracts increased in a concentration-dependent manner. S extract was found to possess higher reducing power than equivalent amount of ascorbic acid at 20 and 25 μg/ml concentrations. Linear correlation between reducing power and concentration of E, A, and W extracts (R² > 0.95) was observed. A, W, and S extracts contained relatively high levels of proline. The results presented suggest that D. hastata may provide a natural source of antioxidants and proline.