379 Journalof Food Protection, Vol. 64, No. 3, 2001, Pages 379– 384 Copyright q,International Association forFood Protection AntioxidantActivity ofResveratrol Compared withCommon FoodAdditives M.ANTONIA MURCIA* AND MAGDALENA MARTI´NEZ-TOME´ Departmentof Food Science, VeterinaryFaculty, Campus of Espinardo,University ofMurcia,Apartado de Correos 4021, E-30080-Murcia, Spain MS00-238: Received 24July 2000/ Accepted 2October2000 ABSTRACT Downloaded from http://meridian.allenpress.com/jfp/article-pdf/64/3/379/2007778/0362-028x-64_3_379.pdf by guest on 29 September 2021 Resveratrolis a phenoliccompound of thestilbene family present in wines and various parts of the grape, including the skin.In this study, the antioxidant and prooxidant activities of resveratrol were compared with other antioxidants (butylated hydroxytoluene[BHT], butylatedhydroxyacetone [BHA], phenol,propyl gallate [PG], sodium tripolyphosphate [TPP], a- tocopherol,and vanillin) widely used in foods. The ability of these compounds to inhibit lipid peroxidation was as follows: BHA . resveratrol . PG . tripolyphosphate . vanillin . phenol . BHT . a-tocopherol,the rstthree inhibiting the peroxidationin a concentration-dependentmanner .Theorder of OH scavengeractivity of the tested compounds was BHA . TPP . BHT.Resveratroland vanillin produced between 10 to 7% and 16 to 10% inhibition of the deoxyribose attack, respectively,but they do not scavenge OH .Neitherthe resveratrol analyzed nor PG orthe rest of compounds reacted with H2O2 andmust be considered inef cient in catalyzing any subsequent oxidation. The ability to scavenge HOCl was,in decreasingorder, PG . resveratrol . a-tocopherol . phenol.The other compounds did not scavenge HOCl. Anantioxidant may be de ned as ‘ ‘asubstancethat beenproposed to have antioxidant capacities include tea, whenpresent at low concentrations compared with those of olives,grapes, apple, clove, Vanillaplanifolia, among oth- anoxidizablesubstrate such as fats,proteins, carbohydrates, ers. The avonoidsand other polyphenols found in some orDNA, signicantly delays or prevents the oxidation of ofthese extracts have been widely discussed as potential thesubstrate’ ’ (1). antioxidantprophylactics (3). Therole of avonoidsin the Antioxidantsare of interestto food scientists, polymer dietis receiving increasing interest and the biological ac- chemists,radiation chemists, and food manufacturers (5). tivityof these polyphenolic components is of great impor- Oxidationof thelipid components in food by the free rad- tancefor understandingtheir mode of action in vivo. In- icalchain reaction of lipidperoxidation is amajorproblem teresthas focused on the health bene ts derived from adiet for foodmanufacturers. The extent to which oxidation of richin avonoids (3). fattyacids and their esters occurs in foods depends on the Amongthe most frequently used antioxidants in food chemicalstructure of thefatty acid, the nature of foodpro- issodium tripolyphosphate (TPP ,E-452)that is used as a cessing,and the temperature at which the foods are stored preservative,metal ion sequestrant, and texturizer in meat and/orcooked, and the minor constituent antioxidants (2). and shproducts (26, 27). BHA (E-320)and BHT (E-321) Manyfood antio xidants(butyl atedhydrox yanisole havebeen used as foodantioxidants because the end-prod- [BHA], butylatedhydroxytoluene [BHT], andpropyl gal- uctsof lipid peroxidation are toxic and can impart a ran- late[PG]) arechain-breaking inhibitors of lipid peroxida- cidityor off- avor to foods like cakes, cookies, vegetable tionthat act through a mechanismthat produces a reaction oils,and dehydrated soups (5). Propylgallate (E-310) has betweenthe radicals and proteins or fatty-acid side-chains beenused as a foodantioxidant, e.g., in vegetableoils, mar- (21). Theuse of these compounds is increasingly limited garine,instant mashed potato, and breakfast cereals. V anil- outof safety considerations, and their replacement by nat- linis also a widelyused avoringagent, e.g., in icecream, uralantioxidants is widely advocated (1). Also,food man- cakes,syrups, and chocolate (4, 11). a-Tocopherol(E-307) ufacturersare very interested in natural antioxidants to act isthemost important natural antioxidant found in vegetable asreplacements for thesynthetic antioxidants currently oil-derivedfoods. It hasa widelyextended use in fats and usedby the industry to combat the formation of off-avors (23). andrancidity, particularly in fat-based products (1, 15). oils a-Tocopherolcontains a phenolicstructure that Thereis an emergingconsensus that the micronutrients scavengeslipid and oxygen radicals (28). andnonnutrient components of fruits and vegetables play Resveratrol(3,4 9,5-trihydroxystilbene) isa phenolic apreventiverole in the development of chronic diseases. compoundof the stilbene family together with other hy- Epidemiologicalstudies have found that diets high in fruits, droxylatecompounds. Resveratrol is found in winesand in vegetables,herbs, and spices correlate with low incidences variousparts of the grape, including the skin (38). Wines ofcancerand heart disease (29, 30). Plantextracts that have from variousparts of the world have been shown to vary widelyin their resveratrol content, on account of thein u- *Authorfor correspondence. E-mail: [email protected]. enceof soil and climate factors (12). Anumberof research- 380 MURCIA AND MARTI´NEZ-TOME´ J.FoodProt., Vol. 64, No. 3 ers havemeasured the resveratrol concentration of wines incubationperiod, 0.1 ml of 2%(wt/vol)BHT wasadded to each (9, 13) anddiscussed the differences in termsof the various mixturefollowed by addition of 1 mleach of 1% (wt/ vol)thio- grapevarieties used (14,19, 34). Thephenolic content of barbituricacid (TBA) and2.8% (wt/ vol)trichloroacetic acid. The redwine is about 20 to 50 times higher than that of white solutionswere heated in a waterbath at 80 8Cfor20 min to de- wine (22). velopthe malondialdehyde thiobarbituric adduct ([TBA] 2-MDA). Theaddition of BHT tothe reaction mixtures minimizes erroneous Inrecent years, many papers have been published on increasesin color due to iron ion-dependent hydroperoxide de- resveratrolin grapes and wines, re ecting the widespread compositionduring the acid heating stage (29). The (TBA)2-MDA interestthis compound has aroused in connection with chromogenwas extracted into 2 mlof butan-1-ol and the extent pharmacologicalproperties that would seem to help prevent ofperoxidation measured in the organic layer as absorbance at cardiovasculardisease. Resveratrol would seem to have an- 532 nm. tiplateletaggregating properties and facilitate an increasein ThisTBA testmeasures not only peroxidation occurring in high-densitylipoprotein cholesterol. Moreover some au- theexperiment itself but also peroxidation taking place during the thorshave recently reported that resveratrol delays the onset acid-heatingstage of theassay. In order to avoidany interference, oftumors in animals. It is known that the moderate con- theTBA testwas performed in the presence of the antioxidant Downloaded from http://meridian.allenpress.com/jfp/article-pdf/64/3/379/2007778/0362-028x-64_3_379.pdf by guest on 29 September 2021 sumptionof redwine reduces ADP-induced platelet aggre- BHT toinhibit peroxidation during the assay itself (6). gation (12). Frankeland coworkers (18) andEctor et al. Hydroxylradical scavenging. Ina nalvolume of 1.2 ml, (16) attributedthe positive bene ts ofredwine to theability reactionmixtures contained the following reagents at the nal ofphenolic substances to prevent oxidation of low-density concentrationstated of 10 mM KH 2PO4-KOH buffer(pH 7.4),2.8 lipoprotein,a criticalevent in the process of atherogenesis mM H2O2,2.8mM deoxyribose(where used), 50 mM FeCl3 pre- (22). Inaddition, investigators concluded that the bene cial mixedwith 100 mMEDTA beforeaddition to the reaction mix- effectsof the beverages, such as beer and red wine, are ture,and 0.05, 0.125, 0.25, and 0.5 mM ofresveratroland the rest partiallydue to components other than the alcohol (37). ofthe compounds tested dissolved in water .Ascorbate(100 mM), whereused, was added to start the reaction. The tubes were in- Neverthelessit should always be takeninto account that the cubatedat 37 8Cfor1 h.The products of hydroxyl radical (OH ) problemsof alcoholism might be a greaterproblem than attackupon deoxyribose were measured as described in Aruoma anyof the bene ts derived from thementioned alcoholic et al. (7). beverages. Theresults of epidemiologicalstudies based on W orld Scavengingof hydrogen peroxide. Resveratrol,PG, andthe HealthOrganization data indicate that the consumption of differentcomponents to be tested for their reaction with H 2O2 wereincubated at 0.05, 0.125, 0.25, and 0.5 mM with6.72 mM wine,especially red wine, may reduce the risk of coronary H O for10 min at 25 8C.Aliquotsof thesecompounds were then heartdisease by 40% (33). 2 2 takenand assayed for remaining H 2O2 byusing the peroxidase Resveratrolis a naturalproduct that has received at- system (6). Theremaining H 2O2 wasmeasured as the formation tentionfor useas a possiblefood antioxidant. The aim of ofa browncolor (recorded at 436 nm) in reaction mixtures con- thisstudy is to compare the in vitro effect of resveratrol taining,in a nalvolume of 1 ml,0.15 M KH 2PO4-KOH buffer, withother widely used antioxidants, such as BHT ,BHA, pH 7.4, 50 mlguaiacolsolution (made by adding 100 ml of pure phenol,PG, TPP, a-tocopherol,and vanillin to better un- liquidto 100 ml water), and 10 mlofSigma type IV horseradish derstandthe nature