İlhami Gülçin

Antioxidant activity of food constituents: an overview

Recently, there has been growing interest in research into the role of plant-derived antioxidants in food and human health. The beneficial influence of many foodstuffs and beverages including fruits, vegetables, tea, coffee, and cacao on human health has been recently recognized to originate from their antioxidant activity. For this purpose, the most commonly methods used in vitro determination of antioxidant capacity of food constituents are reviewed and presented. Also, the general chemistry underlying the assays in the present paper was clarified. Hence, this overview provides a basis and rationale for developing standardized antioxidant capacity methods for the food, nutraceutical, and dietary supplement industries. In addition, the most important advantages and shortcomings of each method were detected and highlighted. The chemical principles of these methods are outlined and critically discussed. The chemical principles of methods of 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonate) radical (ABTS·+) scavenging, 1,1-diphenyl-2-picrylhydrazyl (DPPH·) radical scavenging, Fe3+–Fe2+ transformation assay, ferric reducing antioxidant power (FRAP) assay, cupric ions (Cu2+) reducing power assay (Cuprac), Folin-Ciocalteu reducing capacity (FCR assay), peroxyl radical scavenging, superoxide anion radical (O2·−) scavenging, hydrogen peroxide (H2O2) scavenging, hydroxyl radical (OH·) scavenging, singlet oxygen (1O2) quenching assay and nitric oxide radical (NO·) scavenging assay are outlined and critically discussed. Also, the general antioxidant aspects of main food components were discussed by a number of methods which are currently used for detection of antioxidant properties food components. This review consists of two main sections. The first section is devoted to main components in the foodstuffs and beverages. The second general section is some definitions of the main antioxidant methods commonly used for determination of antioxidant activity of components in the foodstuffs and beverages. In addition, there are given some chemical and kinetic basis and technical details of the used methods.

Screening of antioxidant and antimicrobial activities of anise (Pimpinella anisum L.) seed extracts

In this study, antioxidant and antimicrobial activities of water and ethanol extracts of anise (Pimpinella anisum L.) seed (PAS) were investigated. The antioxidant properties of both extracts of PAS were evaluated using different antioxidant tests, including reducing power, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. Twenty μg/ml of water and ethanol extracts exhibited 99.1 and 77.5% inhibition of peroxidation in linoleic acid system, which was greater than the same concentration of α-tocopherol (36.1%). These various antioxidant activities were compared with synthetic antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and α-tocopherol. The water extract of PAS exhibited greater antioxidant capacity than that of ethanol. Antimicrobial activity tests were carried out using disc diffusion methods with 10 microbial species.

Determination of antioxidant activity of lichen Cetraria islandica (L) Ach.

The study was aimed at evaluating the antioxidant activity of aqueous extract of C. islandica. The antioxidant activity, reducing power, superoxide anion radical scavenging and free radical scavenging activities were studied. The antioxidant activity increased with the increasing amount of extracts (from 50 to 500 microg) added to linoleic acid emulsion. About 50, 100, 250, and 500 microg of aqueous extract of C. islandica showed higher antioxidant activity than 500 microg of alpha-tocopherol. The samples showed 96, 99, 100, and 100% inhibition on peroxidation of linoleic acid, respectively. On the other hand, the 500 microg of alpha-tocopherol showed 77% inhibition on peroxidation on linoleic acid emulsion. Like antioxidant activity, the reducing power, superoxide anion radical scavenging and free radical scavenging activities of C. islandica depends on concentration and increasing with increased amount of sample. The results obtained in the present study indicate that C. islandica is a potential source of natural antioxidant.

Antioxidant and analgesic activities of turpentine of Pinus nigra Arn. subsp. pallsiana (Lamb.) Holmboe

The aim of this study is to examine possible antioxidant and analgesic activities of turpentine exudes from Pinus nigra Arn. subsp. pallsiana (Lamb.) Holmboe (TPN). Total antioxidant activity, reducing power, superoxide anion radical scavenging, free radical scavenging, metal chelating, and hydrogen peroxide scavenging activities were studied. The total antioxidant activity increased with the increasing amount of extracts (100, 300, and 500 microg) added to linoleic acid emulsion. All of the doses of TPN showed higher antioxidant activity than alpha-tocopherol. The samples showed 49, 70, and 91% inhibition on peroxidation of linoleic acid emulsion, respectively. On the other hand, the 300 microg of alpha-tocopherol showed 40% inhibition on peroxidation of linoleic acid emulsion. There is correlation between antioxidant activity and the reducing power, superoxide anion radical scavenging, free radical scavenging, metal chelating, and hydrogen peroxide scavenging activities. Like antioxidant activity, the reducing power, superoxide anion radical scavenging, free radical scavenging, metal chelating, and hydrogen peroxide scavenging activities of TPN depending on concentration and increasing with increased concentration of TPN. These properties may be the major reasons for the inhibition of lipid peroxidation. The results obtained in the present study indicate that the TPN has a potential source of natural antioxidant. In addition, analgesic effect of TPN was investigated in present study and TPN had strong analgesic effect. The analgesic effect of TPN compared with metamizol as a standard analgesic compound.

Carbonic anhydrase inhibitors. Inhibition of human erythrocyte isozymes I and II with a series of antioxidant phenols

The inhibition of two human cytosolic carbonic anhydrase (hCA, EC 4.2.1.1) isozymes I and II, with a series of phenol derivatives was investigated by using the esterase assay, with 4-nitrophenyl acetate as substrate. 2,6-Dimethylphenol, 2,6-diisopropylphenol (propofol), 2,6-di-t-butylphenol, butylated hydroxytoluene, butylated hydroxyanisole, vanillin, guaiacol, di(2,6-dimethylphenol), di(2,6-diisopropylphenol), di(2,6-di-t-butylphenol), and acetazolamide showed K(I) values in the range of 37.5-274.5 microM for hCA I and of 0.29-113.5 microM against hCA II, respectively. All these phenols were non-competitive inhibitors with 4-nitrophenylacetate as substrate. Some antioxidant phenol derivatives investigated here showed effective hCA II inhibitory effects, in the same range as the clinically used sulfonamide acetazolamide, and might be used as leads for generating enzyme inhibitors possibly targeting other CA isoforms which have not been yet assayed for their interactions with such agents.

On the in vitro antioxidative properties of melatonin

Abstract:The aim of this study is to examine possible in vitro antioxidant effects of melatonin. Thus, the total in vitro antioxidant activity of melatonin was studied using a thiocyanate method. Additionally, the reducing power, the superoxide anion scavenging activity and free radical scavenging activity of melatonin were determined. Melatonin exhibited potent antioxidant activity in a linoleic acid emulsion system. The antioxidant activity increased with increasing concentrations of melatonin (50–500 μg). The 50, 100, 250 and 500 μg melatonin doses showed 41, 60, 86 and 99% inhibition of peroxidation of linoleic acid, respectively. On the other hand, a 500‐μg dose of α‐tocopherol showed 34% inhibition of peroxidation of linoleic acid. Like the total antioxidant activity, the reducing power of melatonin increased in a dose‐dependent manner. The reducing power of melatonin was statistically significant versus control, but lower than butylated hydroxytoluene (BHT) or quercetin. Additionally, melatonin had potent superoxide radical scavenging activity and exhibited a higher superoxide radical scavenging activity than quercetin or BHT but lower than butylated hydroxyanisole (BHA). Melatonins direct free radical scavenging actions may account, at least in part, for its ability to reduce lipid peroxidation. Melatonin may have utility in protecting stored foods from free radical‐induced deterioration.

The antioxidant and radical scavenging activities of black pepper (Piper nigrum) seeds

Water and ethanol crude extracts from black pepper (Piper nigrum) were investigated for their antioxidant and radical scavenging activities in six different assay, namely, total antioxidant activity, reducing power, 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. Both water extract (WEBP) and ethanol extract (EEBP) of black pepper exhibited strong total antioxidant activity. The 75 µg/ml concentration of WEBP and EEBP showed 95.5% and 93.3% inhibition on peroxidation of linoleic acid emulsion, respectively. On the other hand, at the same concentration, standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and α-tocopherol exhibited 92.1%, 95.0%, and 70.4% inhibition on peroxidation of linoleic acid emulsion, respectively. Also, total phenolic content in both WEBP and EEBP were determined as gallic acid equivalents. The total phenolics content of water and ethanol extracts were determined by the Folin-Ciocalteu procedure and 54.3 and 42.8 µg gallic acid equivalent of phenols was detected in 1 mg WEBP and EEBP.

Carbonic anhydrase inhibitors. Inhibition of mammalian isoforms I–XIV with a series of natural product polyphenols and phenolic acids

A series of phenolic acids and phenol natural products, such as p-hydroxybenzoic acid, p-coumaric acid, caffeic acid, ferulic acid, gallic acid, syringic acid, quercetin, and ellagic acid, were investigated for their inhibitory effects against the metalloenzyme carbonic anhydrase (CA, EC 4.2.1.1). All mammalian isozymes of human (h) or murine (m) origin hCA I-hCA XII, mCA XIII and hCA XIV were inhibited in the low micromolar or submicromolar range by these (poly)phenols (K(I)s in the range of 0.87-7.79 microM). p-Hydroxybenzoic acid was the best inhibitor of all isozymes (K(I)s of 0.87-35.4 microM) and the different isozymes showed very variable inhibition profiles with these derivatives. Phenols like the ones investigated here possess a CA inhibition mechanism distinct of that of the sulfonamides/sulfamates used clinically or the coumarins. Unlike the sulfonamides, which bind to the catalytic zinc ion, phenols are anchored at the Zn(II)-coordinated water molecule and bind more externally within the active site cavity, making contacts with various amino acid residues. As this is the region with the highest variability between the many CA isozymes found in mammals, this class of compounds may lead to isoform-selective inhibitors targeting just one or few of the medicinally relevant CAs.

Metal chelating and hydrogen peroxide scavenging effects of melatonin.

Abstract: Antioxidant activity of a molecule is attributed to various mechanisms such as prevention of chain initiation, binding of transition metal ion catalysts and decomposition of peroxides. This study was aimed at evaluating the metal chelating and hydrogen peroxide (H2O2) scavenging activity of melatonin. The metal chelating and H2O2 scavenging activity increased with increasing concentrations of melatonin (20–60 μg/mL). α‐Tocopherol, butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) were used as standards. Sixty micrograms per milliliter concentration of melatonin exhibited 95% chelating effect on ferrous ions and scavenged 83% of H2O2. On the other hand, the same concentration of α‐tocopherol, BHA, and BHT exhibited 58, 61, and 72% inhibition, respectively, of the formation of the Fe2+–ferrozine complex and scavenged 48, 20, and 23%, respectively, of H2O2. Based on these results, it is concluded that melatonin is an effective metal chelating agent and scavenger of H2O2. These properties may be major reasons for the melatonins ability to inhibit lipid peroxidation.

In Vitro Inhibition of Human Carbonic Anhydrase I and II Isozymes with Natural Phenolic Compounds

Inhibition of two human cytosolic carbonic anhydrase (hCA, EC 4.2.1.1) isozymes I and II with some natural phenolic derivatives was investigated using the esterase assay with 4‐nitrophenyl acetate as substrate. Resveratrol, catechin, silymarin, dobutamin, and curcumin showed KI values in the range of 4.47–9.47 mm for hCA I and of 2.86–7.44 μm against hCA II, respectively. These natural product phenols were generally competitive inhibitors with 4‐nitrophenylacetate as substrate. Some natural phenols investigated here showed effective hCA II inhibitory effects, in the same range as the clinically used sulfonamide acetazolamide, and might be used as leads for generating enzyme inhibitors possibly targeting other CA isoforms that have not been yet assayed for their interactions with such agents.