1. bookVolume 12 (2019): Issue 2 (October 2019)
Journal Details
First Published
10 Dec 2012
Publication timeframe
2 times per year
access type Open Access

Selectedin vitro methods to determine antioxidant activity of hydrophilic/lipophilic substances

Published Online: 21 Jan 2020
Volume & Issue: Volume 12 (2019) - Issue 2 (October 2019)
Page range: 200 - 211
Journal Details
First Published
10 Dec 2012
Publication timeframe
2 times per year

The topic of free radicals and related antioxidants is greatly discussed nowadays. Antioxidants help to neutralize free radicals before damaging cells. In the absence of antioxidants, a phenomenon called oxidative stress occurs. Oxidative stress can cause many diseases e.g. Alzheimer’s disease and cardiovascular diseases. Therefore, antioxidant activity of various compounds and the mechanism of their action have to be studied. Antioxidant activity and capacity are measured by in vitro and in vivo methods; in vitro methods are divided into two groups according to chemical reactions between free radicals and antioxidants. The first group is based on the transfer of hydrogen atoms (HAT), the second one on the transfer of electrons (ET). The most frequently used methods in the field of antioxidant power measurement are discussed in this work in terms of their principle, mechanism, methodology, the way of results evaluation and possible pitfalls.


Abramovič H, Grobin B, Ulrih NP, Cigić B (2018) Journal of Chemistry 2018: 9 pages.10.1155/2018/4608405Search in Google Scholar

Ainsworth EA, Gillespie KM (2007) Nature Protocols 2 (4): 875—877.10.1038/nprot.2007.102Search in Google Scholar

Akar Z, Burnaz NA (2019) LWT 112: 108212.10.1016/j.lwt.2019.05.110Search in Google Scholar

Alam MN, Bristi NJ, Rafiquzzaman M (2013) Saudi Pharmaceutical Journal 21 (2): 143—152.10.1016/j.jsps.2012.05.002Search in Google Scholar

Al-Farsi M, Al-Amri A, Al-Hadhrami A, Al-Belushi S (2018) Heliyon 4 (10): e00874.10.1016/j.heliyon.2018.e00874Search in Google Scholar

Alshaal S, Karabet F, Daghestani M (2019) Analytical and Bioanalytical Chemical Research 6: 97—110.Search in Google Scholar

Apak R, Capanoglu E, Shahidi F (2018) Measurement of Antioxidant Activity and Capacity: Recent Trends and Applications. John Wiley & Sons, USA.10.1002/9781119135388Search in Google Scholar

Apak R, Güçlü K, Özyürek M, Celik SE (2008) Microchemica Acta 160 (4): 413—419.10.1007/s00604-007-0777-0Search in Google Scholar

Askin H, Yilmaz B, Gulcin I, Taslimi P, Bulut S, Yildiz M, Kandemir N (2018) Indian Journal of Pharmaceutical Sciences 80 (5): 802—812.10.4172/pharmaceutical-sciences.1000425Search in Google Scholar

Behzadnezhad B, Behdad N, Mcmillan AB (2017) United States Patent Application Publication 2018/0340999 A1.Search in Google Scholar

Benzie IFF, Strain JJ (1999) Methods in Enzymology 299: 15—27.10.1016/S0076-6879(99)99005-5Search in Google Scholar

Berker KI, Ozdemir Olgun FA, Ozyurt D, Demirata B, Apak R (2013) Journal of Agricultural and Food Chemistry 61 (20): 4783—4791.10.1021/jf400249k23627440Search in Google Scholar

Borbat PP, Costa-Filho AJ, Earle KA, Moscicki JK, Freed JH (2001) Science 291: 266—269.10.1126/science.291.5502.26611253218Search in Google Scholar

Bortolomeazzi R, Sebastianutto N, Tonioolo R, Pizzariello A (2007) Food Chemistry 100 (4): 1481—1489.10.1016/j.foodchem.2005.11.039Search in Google Scholar

Bukhari MH, Batool S, Raza DY, Bagasra O, Rizvi A, Shah A, Sultan T (2018) Electromagnetic Biology and Medicine 37 (3): 1—6.10.1080/15368378.2018.149903230024785Search in Google Scholar

Burlaka AP, Vovk AV, Burlaka AA, Gafurov MR, Iskhakova KB, Lukin SN (2018) BioMed Research International: 4848652.10.1155/2018/4848652611000630186860Search in Google Scholar

Burnaz NA, Küçük M, Akar Z (2017) Journal of Chromatography B, 1052: 66—72.10.1016/j.jchromb.2017.03.00328359985Search in Google Scholar

Celik SE, Asfoor A, Apak R (2019) Journal of Agricultural and Food Chemistry 67 (29): 8279—8289.10.1021/acs.jafc.9b0300131259549Search in Google Scholar

Chiavaroli V, Giannini C, De Marco S, Chiarelli F, Mohn A (2011) Redox Report 16: 101—107.10.1179/174329211X13049558293551683764721801491Search in Google Scholar

Choy CKM, Benzie IFF, Cho P (2000) Biochemistry and Molecular Biology 41 (11): 3293—3298.Search in Google Scholar

Davies MJ ((2016) Methods 109: 21—30.10.1016/j.ymeth.2016.05.01327211009Search in Google Scholar

Denardin CC, Hirsch GE, Rocha RF, Vizzotto M, He nriques AT, Moreira JCF, Guma F, Emanueloli T (2015) Journal of Food and Drug Analysis 23 (3): 387—398.10.1016/j.jfda.2015.01.00628911695Search in Google Scholar

Drouet S, Doussot J, Garros L, Mathiron D, Bassard S, Favre-Réguillon, Molinié R, Lainé Éric, Hano Christophe (2018) Molecules 23 (10): 2594.10.3390/molecules23102594622264430309022Search in Google Scholar

Everette JD, Bryant QM, Green AM, Abbey Y, Wangila GW, Walker R (2010) Journal of Agricu lture and Food Chemistry 58 (14): 8139—8144.10.1021/jf1005935Search in Google Scholar

Farajzadeh M, Abbaspour MA, Mogaddama MRA, Mogaddam A, Alizadeh Nabil AAAN (2016) Analytical and Bioanalytical Chemical Research 3: 239—251.Search in Google Scholar

Ferrante AA, Martins IS, Silva LA, Percário S, Ferreira MES (2019) Australian Journal of Basic and Applied Sciences 13 (3): 19—22.Search in Google Scholar

Floegel A, Kim D, Chung SJ, Koo SI, Chun OK (2011) Journal of Food Composition and Analysis 24: 1043—1048.10.1016/j.jfca.2011.01.008Search in Google Scholar

Franzoni F, Federighi G, Fusi J, Agosta V, Cerri E, Banducci R, Petrocchi A, Bernardi R, Innocenti A, Pruneti C, Daniele S, Pellegrini S, Martini C, Scuri R, Galetta F (2017) Archives Italiennes de Biologe 155: 1—10.Search in Google Scholar

Gardner PT, McPhail DB, Crozier A, Duthie GG (1999) Journal of Science and Food Agriculture 79: 1011—1014.10.1002/(SICI)1097-0010(19990515)79:7<1011::AID-JSFA320>3.0.CO;2-YSearch in Google Scholar

Garrett AR, Murray BK, Robison RA, O’Neill KL (2010) Advanced protocols in oxidative stress 2: 251—262.10.1007/978-1-60761-411-1_17Search in Google Scholar

Goosen NJ, Ooshuizen D, Stander MA, Dabai AI, Pedavoah MM, Usman GO (2018) South African Journal of Botany 119: 11—16.10.1016/j.sajb.2018.08.008Search in Google Scholar

Granato D, Shahidi F, Wrolstad R, Kilmartin P, Melton LD, Hidalgo FJ, Miyashita K, Camp J, Alasalvar C, Ismail AB, Elmore S, Birch GG, Astley SB, Pegg R, Zhou P, Finglas P (2018) Food Chemistry 264: 471—475.10.1016/j.foodchem.2018.04.012Search in Google Scholar

Gupta D (2015) International Journal of Pharmaceutical Sciences and Research 6 (2): 546—66.Search in Google Scholar

Haida Z, Hakiman M (2019) Food Science & Nutrition 7 (5): 1555—1563.10.1002/fsn3.1012Search in Google Scholar

Holtz RW (2009) Skin Aging Handbook: In Vitro Methods to Screen Materials for Anti-aging Effects. William Andrew, Norwich NY.10.1016/B978-0-8155-1584-5.50017-XSearch in Google Scholar

Huang D, Ou B, Prior RL (2005) Journal of Agricultural and Food Chemistry 53(6): 1841.10.1021/jf030723cSearch in Google Scholar

Huda-Faujan N, Norrakiah AS, Babji AS (2009) African Journal of Biotechnology 8 (3): 484—489.Search in Google Scholar

Jiang J, Zang S, Li D, Wang K, Tian S, Yu A, Zhang Z (2018) Talanta 184: 23—28.10.1016/j.talanta.2018.02.098Search in Google Scholar

Jiang Y, Rakesh KP, Alharbi NS, Vivek HK, Manukumar HM, Mohammed YHE, Qin H (2019) Bioorganic Chemistry 89: 103015.10.1016/j.bioorg.2019.10301531158576Search in Google Scholar

Joseph NM, Sabharwal M, Shashi A, Mahor A, Rawal S (2018) International Journal of Pharmaceutical Sciences and Research 1: 1—11.Search in Google Scholar

Kamer G, Demirata B, Bayraktar R, Ozyurt D, Apak R (2019) Analytical Methods 14.Search in Google Scholar

Kedare SB, Singh RP (2011) Journal of Food Science and Technology 48 (4): 412—422.10.1007/s13197-011-0251-1355118223572765Search in Google Scholar

Kimáková T, Baranovičová I (2015) Léčivá síla antioxidantú. Príroda, s.r.o, Bratislava.Search in Google Scholar

Klaunig JE, Wang Z (2018) Current Opinion in Toxicology 7: 116—121.10.1016/j.cotox.2017.11.014Search in Google Scholar

Kleschyov AL, Wenzel P, Munzel T (2007) Journal of Chromatography B: 851 (1, 2), 12—20.10.1016/j.jchromb.2006.10.00617070113Search in Google Scholar

Koleva II, Niederländer H, Beek TA (2000) Analytical Chemistry 73 (14): 3373—3381.10.1021/ac001361011476238Search in Google Scholar

Lachowicz S, Oszmiański J, Wiśniewski R (2018) European Food Research and Technology 244: 1269—1280.10.1007/s00217-018-3042-3Search in Google Scholar

Lamuela-Raventós RM (2018) Measurement of Antioxidant Activity and Capacity: Recent Trends and Applications. John Wiley & Sons, USA.Search in Google Scholar

Li D, Jiang J, Han D, Yu X, Wang K, Zang S, Lu D, Yu A, Zhang Z (2016) Analytical Chemistry 88: 3885—3890.10.1021/acs.analchem.6b0004926927869Search in Google Scholar

Li XY, Wang ZY, Wang L, Walid E, Zhang H (2012) International Journal of Molecular Science 13 (5): 5801—5817.10.3390/ijms13055801338280722754332Search in Google Scholar

Lichtenthäler R, Marx F (2003) European Food Research Technology 216: 166—173.10.1007/s00217-002-0635-6Search in Google Scholar

Lim S, Choi A-H, Kwon M, Joung E, Shin T, Lee S, Kim N, Kim H (2019) Food Chemistry 278: 178—184.10.1016/j.foodchem.2018.11.05830583359Search in Google Scholar

Lobo V, Patil A, Chandra N (2010) Pharmacognosy Reviews 4 (8): 118—126.10.4103/0973-7847.70902Search in Google Scholar

Ma H, Liu T, Li J, Mingya D, Gao X, Chang Y (2019) Journal of Chromatography A 1593: 147—155.10.1016/j.chroma.2019.01.050Search in Google Scholar

Martín J, Kuskoski EM, Navas MJ, Asuero AG (2017) Flavonoids — From Biosynthesis to Human Health: Antioxidant Capacity of Anthocyanin Pigments. BoD — Books on Demand, Germany.10.5772/67718Search in Google Scholar

Mellado-Ortega E, Zabalgogeazcoa I, Aldana B, Arellano JB (2017) Analytical Biochemistry 519: 27—29.10.1016/j.ab.2016.12.009Search in Google Scholar

Minussi RC, Rossi M, Bologna L, Cordi L, Rotilio D, Pastore G, Durán N (2003) Food Chemistry 82 (3): 409—416.10.1016/S0308-8146(02)00590-3Search in Google Scholar

Moran-Pa lacio EF, Zamora-Álvarez LA, Stephens-Camacho NA, Yánez-Farías GA, Virgen-Ortiz A, Martínez-Cruz O, Rosas-Rodríguez JA (2014) Tropical Journal of Pharmaceutical Research 13 (9): 1487—1493.10.4314/tjpr.v13i9.15Search in Google Scholar

Moukette BM, Pieme CA, Njimou JR, Biapa CP, Marco B, Ngogang JY (2015) Biological Research 48(1): 15.10.1186/s40659-015-0003-1439359025885269Search in Google Scholar

Munialo CD, Naumovski N, Sergi D, Stewart D, Mellor DD (2019) 54 (5): 1448—1459.10.1111/ijfs.14135Search in Google Scholar

Murauer A, Bakry R, Schottenberger H, H uck C, Ganzera M (2017) Analytica Chimica Acta 963: 136—142.10.1016/j.aca.2017.01.048538818128335967Search in Google Scholar

Nijhawan P, Arora S (2019) Obesity Medicine 15: 100—125.10.1016/j.obmed.2019.100125Search in Google Scholar

Obón JM, Castellar R, Cascales JA, Fernándey-Lópey JA (2005) Food Research International 38 (8): 843—845.10.1016/j.foodres.2005.01.010Search in Google Scholar

Ojha K, Dubey S, Chandrakar J, Minj RA, Dehariya R, Dixit AK (2018) Research Journal of Life Sciences, Bioinformatics, Pharmaceutical and Chemical Sciences 4 (6): 707.Search in Google Scholar

Okazaki S, Takeshita K (2018) Applied Magnetic Resonance 49 (8): 881—892.10.1007/s00723-018-0999-9Search in Google Scholar

Ononamadu C, Ihegboro GO, Owolarafe K, Salawu K, Fadilu M, Eyeigwe OC, Oshobu ML, Nwachukwu FC (2019) Analytical and Bioanalytical Chemical Research 6: 431—439.Search in Google Scholar

Ordoudi SA, Tsimidou MZ (2006) Journal of Agricultural and Food Chemistry 54 (5): 1663—1671.10.1021/jf052731uSearch in Google Scholar

Oribayo OO, Owolabi MA, Ukpo GE, Shode FO (2018) Tropical Journal of Natural Product Research 2 (1): 18—22.10.26538/tjnpr/v2i1.4Search in Google Scholar

Pérez-Burillo S, Rufián-Henares JA, Pastoriza S (2018) Food Chemistry 239: 1263—1272.10.1016/j.foodchem.2017.07.063Search in Google Scholar

Pérez-Jiménez J, Saura-Calixto F (2008) International Journal of Food Science & Technology 43 (1): 185—191.10.1111/j.1365-2621.2006.01425.xSearch in Google Scholar

Pisoschi AM, Negulescu G (2011) Biochemistry & Analytical Biochemistry 1 (1).Search in Google Scholar

Prieto MA, Vázquez JA, Murado M (2015) Food Chemistry 167 (15): 299—310.10.1016/j.foodchem.2014.06.114Search in Google Scholar

Prior RL (2015) Journal of Functional Foods 18 (B): 797—810.10.1016/j.jff.2014.12.018Search in Google Scholar

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999) Free Radical Biology and Medicine 26 (9, 10): 1231—1237.10.1016/S0891-5849(98)00315-3Search in Google Scholar

Regoli F (2000) Aquatic Toxicology 50 (4): 351—361.10.1016/S0166-445X(00)00091-6Search in Google Scholar

Rodriguez-Nogales J, Vila-Crespo J, Gómez M (2011) Food Chemistry 129: 1800—1805.10.1016/j.foodchem.2011.05.105Search in Google Scholar

Sakurai Y, Yamaguchi T, Ando K, Kuwabara K, Nakajima H, Morimoto C, Yoshikawa S, Niwa M, Miyake Y, Kanaori K, Nakajim A, Tajima K (2019) Bulletin of Chemical Society of Japan 92 (7): 1218—1225.10.1246/bcsj.20190059Search in Google Scholar

Sánchez-Rangel JC, Benavides J, Heredia JB, Cisneros-Zevallos L, Jacobo-Velázquez (2013) Analytical Methods 5 (21): 5990—5999.10.1039/c3ay41125gSearch in Google Scholar

Schaich KM, Tian X, Xie J (2015) Journal of Functional Foods 18 (B): 782—796.10.1016/j.jff.2015.05.024Search in Google Scholar

Seesom C, Jumepaeng T, Luthria DL, Chanthai S (2018) Journal of Food Health and Bioenvironmental Science 11 (2): 28—37.Search in Google Scholar

Shalaby A (2013) African Journal of Pharmacy and Pharmacology 7 (10): 528—539.10.5897/AJPP2013.3474Search in Google Scholar

Shekhar TC, Anju G (2014) American Journal of Ethnomedicine 1 (4): 244—249.Search in Google Scholar

Shi S, Guo K, Tong R, Liu Y, Tong C, Peng M (2019) Food Chemistry 288: 215—220.10.1016/j.foodchem.2019.03.01330902284Search in Google Scholar

Singh S, Singh RP (2008) Food Reviews International 24 (4): 392—415.10.1080/87559120802304269Search in Google Scholar

Singleton VL, Orthofer R, Lamuela-Raventos RM (1999) Methods Enzymol. 299 : 152—178.Search in Google Scholar

Sotto AD, Checcon P, Celestino I, Locatelli M, Carissimi S, Angelis MD, Rossi V, Limongi D, Toniolo Ch, Martinoli L, Giacomo SD, Palamara AT, Nencioni L (2018) Oxidative Medicine and Cellular Longevity 2018: 1—14.10.1155/2018/5919237608151630140367Search in Google Scholar

Sundararajan R, Ilengesan R (2018) Free Radicals and Antioxidants 8 (1): 55—61.Search in Google Scholar

Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Hawkins Byrne D (2006) Journal of Food Composition and Analysis: 19 (6, 7): 669—675.10.1016/j.jfca.2006.01.003Search in Google Scholar

Torre A, Henderson T, Nigam PS, Owusu-Apenten RK (2015) Food Chemistry 174: 119—123.10.1016/j.foodchem.2014.11.00925529660Search in Google Scholar

Van Den Berg R, Bast A (1999) Food Chemistry 66 (4): 511—517.10.1016/S0308-8146(99)00089-8Search in Google Scholar

Wan HC, Sultana B, Nigam PS, Owusu-Apenten R (2018) Beverages 4 (3): 58.10.3390/beverages4030058Search in Google Scholar

Wayner DD, Burton GW, Ingold KU, Locke S (1985) FEBS Lett. 187: 33—37.Search in Google Scholar

Wong C, Li H, Cheng K, Chen F (2006) Food Chemistry 97 (4): 705—711.10.1016/j.foodchem.2005.05.049Search in Google Scholar

Yadav A, Kumari R, Yadav A, Mishra JP, Srivastava S, Prabha S (2016) Research of Environmental and Life Science 9 (11): 1328—1331.Search in Google Scholar

Yamaguchi F, Yoshimura Y, Nakazawa H, Ariga T (1999) Journal of Agricultural and Food Chemistry 47: 2544—2548.10.1021/jf980676210552524Search in Google Scholar

Yan X, Murphy BT, Hammond GB, Vinson JA, Neto CC (2002) Journal of Agriculture and Food Chemistry 50 (20): 5844—5849.10.1021/jf020223412358448Search in Google Scholar

Yeo JD, Shahidi F (2019) Journal of Agricultural and Food Chemistry 67: 7526—7529.10.1021/acs.jafc.9b0246231184887Search in Google Scholar

Yeum KJ, Russell RM, Krinsky NI, Aldini G (2004) Archives of Biochemistry and Biophysics 430 (1): 97—103.10.1016/j.abb.2004.03.00615325916Search in Google Scholar

Yu LL, Cheng Z (2008) Molecular Nutrition & Food Research 52: 62—78.10.1002/mnfr.20070039518080243Search in Google Scholar

Zablocka A, Sokolowska A, Macala J, Bartoszewska M, Mitkiewicz M, Janusz M, Wilusz T, Polanowski A (2019) International Journal of Peptide Research and Therapeutics: 1—10.Search in Google Scholar

Zang S, Tian S, Jiang J, Han D, Yu X, Wang K, Li D, Lu D, Yu A, Zhang Z (2017) Food Chemistry 221: 1221—1225.10.1016/j.foodchem.2016.11.03627979081Search in Google Scholar

Zhong Y, Shahidi F (2015) Food Science, Technology and Nutrition: 287—333.10.1016/B978-1-78242-089-7.00012-9Search in Google Scholar

Zhong Y, Shahidi F (2015) Handbook of antioxidants for food preservation: methods for the assessment of antioxidant activity in foods. Woodhead Publishing, UK.Search in Google Scholar

Recommended articles from Trend MD

Plan your remote conference with Sciendo