Measurement of Antioxidant Activity and Capacity Recent Trends and Applications

A comprehensive reference for assessing the antioxidant potential of foods and essential techniques for developing healthy food products

Measurement of Antioxidant Activity and Capacity offers a much-needed resource for assessing the antioxidant potential of food and includes proven approaches for creating healthy food products. With contributions from world-class experts in the field, the text presents the general mechanisms underlying the various assessments, the types of molecules detected, and the key advantages and disadvantages of each method. Both thermodynamic (i.e. efficiency of scavenging reactive species) and kinetic (i.e. rates of hydrogen atom or electron transfer reactions) aspects of available methods are discussed in detail. 

A thorough description of all available methods provides a basis and rationale for developing standardized antioxidant capacity/activity methods for food and nutraceutical sciences and industries. This text also contains data on new antioxidant measurement techniques including nanotechnological methods in spectroscopy and electrochemistry, as well as on innovative assays combining several principles. Therefore, the comparison of conventional methods versus novel approaches is made possible. This important resource:

• Offers suggestions for assessing the antioxidant potential of foods and their components
• Includes strategies for the development of healthy functional food products
• Contains information for identifying antioxidant activity in the body
• Presents the pros and cons of the available antioxidant determination methods, and helps in the selection of the most appropriate method

Written for researchers and professionals in the nutraceutical and functional food industries,academia and government laboratories, this text includes the most current knowledge in order to form a common language between research groups and to contribute to the solution of critical problems existing for all researchers working in this field. 

About the editors
Resat Apak is Professor of Analytical Chemistry Division, Department of Chemistry, Faculty of Engineering, Istanbul University, Turkey.

Esra Capanoglu is Associate Professor at the Food Engineering Department, Faculty of Chemical and Metallurgical Engineering, Istanbul Technical University, Turkey.

Fereidoon Shahidi is a University Research Professor at the Department of Biochemistry, Memorial University of Newfoundland, St. John's, Canada.

List of contributors xi

1 Nomenclature and general classification of antioxidant activity/capacity assays 1
Yong Sun, Cheng Yang, and Rong Tsao

1.1 Introduction 1

1.2 Nomenclature of antioxidant activity/capacity assays 2

1.3 Classification of antioxidant activity/capacity assays 2

1.4 Conclusions 15

References 15

2 Assays based on competitive measurement of the scavenging ability of reactive oxygen/nitrogen species 21
Dejian Huang and Restituto Tocmo

2.1 Introduction 21

2.2 Kinetics is more important than thermodynamics when it comes to scavenging ROS 22

2.3 Peroxyl radical scavenging capacity assay based on inhibition of lipid autoxidation 23

2.4 Application of molecular probes for quantification of antioxidant capacity in scavenging specific ROS/RNS 26

2.5 Conclusion: a unified approach for measuring antioxidant capacity against different ROS? 35

Acknowledgment 36

References 36

3 Evaluation of the antioxidant capacity of food samples: a chemical examination of the oxygen radical absorbance capacity assay 39
Eva Dorta, Eduardo Fuentes�]Lemus, Hernan Speisky, Eduardo Lissi, and Camilo Lopez�]Alarcon

3.1 Introduction 39

3.2 Chemical assays to evaluate the antioxidant capacity of food samples 41

3.3 Chemical examination of the ORAC assay: advantages and drawbacks 46

3.4 Future perspectives to improve the antioxidant capacity evaluation of food samples 50

3.5 Conclusions 52

Acknowledgments 52

References 52

4 Electron transfer�]based antioxidant capacity assays and the cupricion reducing antioxidant capacity (CUPRAC) assay 57
Resat Apak

4.1 Introduction 57

4.2 ET�]based TAC assays 58

4.3 CUPRAC assay of antioxidant capacity measurement 64

References 71

5 The ferric reducing/antioxidant power (FRAP) assay for non�]enzymatic antioxidant capacity: concepts, procedures, limitations and applications 77
Iris F.F. Benzie and Malegaddi Devaki

5.1 Introduction: concepts and context 77

5.2 The ferric reducing/antioxidant power (FRAP) assay: a brief overview 79

5.3 Working concepts, what results represent, potential interferences, and limitations 80

5.4 Method outline and detailed procedures for manual, semi�]automated, and fully automated modes 83

5.5 Technical tips for the FRAP assay 89

5.6 Issues of standardization (calibration) and how results are expressed 93

5.7 Issues of sample handling, storage, and extraction 94

5.8 Modifications to the FRAP assay 94

5.9 Illustrative applications 99

5.10 Cautions and concluding remarks 99

Acknowledgments 102

References 102

Further Reading 104

6 Folin–Ciocalteu method for the measurement of total phenolic content and antioxidant capacity 107
Rosa M. Lamuela�]Raventos

6.1 Introduction 107

6.2 Is the Folin–Ciocalteu method an antioxidant assay? 107

6.3 Folin–Ciocalteu assay to quantify phenolic compounds 108

6.4 Folin–Ciocalteu index in wines 109

6.5 Improving the method: more sustainability, less time, and lower cost 110

6.6 Beneficial effects of polyphenols measured by the Folin–Ciocalteu assay in human biological samples: a biomarker of polyphenol intake 114

References 114

7 ABTS/TEAC (2,2′�]azino�]bis(3�]ethylbenzothiazoline�]6�]sulfonic acid)/ TroloxR�]Equivalent Antioxidant Capacity) radical scavenging mixed�]mode assay 117
Antonio Cano and Marino B. Arnao

7.1 Introduction 117

7.2 Use of ABTS as a sensor of antioxidant activity: the TEAC assay 119

7.3 Advantages and disadvantages 125

7.4 TEAC assay in hyphenated and high�]throughput techniques 126

7.5 TEAC in pure compounds 128

7.6 TEAC in foods 130

7.7 Future perspectives 134

References 135

8 DPPH (2,2�]di(4�]tert�]octylphenyl)�]1�]picrylhydrazyl) radical scavenging mixed�]mode colorimetric assay(s) 141
Nikolaos Nenadis and Maria Z. Tsimidou

8.1 Overview 141

8.2 Characteristics of the DPPH radical 142

8.3 The concept behind the development of the DPPH• colorimetric assay 144

8.4 How can antioxidants scavenge the DPPH•? 144

8.5 The evolution of ideas on the underlying mechanism 145

8.6 The DPPH• colorimetric assay(s) 152

8.7 Toward the standardization of a DPPH• assay to address structure–activity relationship issues 154

8.8 Toward the establishment of a DPPH• assay for regulatory and market needs 158

8.9 Concluding remarks – A la recherche du temps perdu 160

References 161

9 Biomarkers of oxidative stress and cellular�]based assays of indirect antioxidant measurement 165
Cheng Yang, Fereidoon Shahidi, and Rong Tsao

9.1 Introduction 165

9.2 Oxidative stress 166

9.3 Biomarkers of oxidative stress 169

9.4 Cell�]based assays of indirect antioxidant measurement 175

9.5 Conclusion 180

References 181

10 Nanotechnology�]enabled approaches for the detection of antioxidants by spectroscopic and electrochemical methods 187
Ryan T. Rauhut, Gonca Bulbul, and Silvana Andreescu

10.1 Introduction 187

10.2 Spectroscopic nano�]based approaches for antioxidant detection 190

10.3 Electrochemical detection 195

10.4 Conclusions and future research needs 200

Acknowledgments 200

References 204

11 Novel methods of antioxidant assay combining various principles 209
Takayuki Shibamoto

11.1 Introduction 209

11.2 Lipid peroxidation and formation of primary and secondary oxidation products 210

11.3 Use of gas chromatography for antioxidant assays 211

11.4 Novel gas chromatographic antioxidant assays 213

11.5 Conclusion 218

References 218

12 Physico�]chemical principles of antioxidant action, including solvent and matrix dependence and interfacial phenomena 225
Katarzyna Jodko�]Piorecka, Jakub Cedrowski, and Grzegorz Litwinienko

12.1 Introduction 225

12.2 Mechanism and kinetics of peroxidation 226

12.3 Initiation of lipid peroxidation chains 227

12.4 Antioxidants 232

12.5 How to recognize a good chain�]breaking antioxidant 234

12.6 Determination of reactivity of a CBA towards peroxyl radicals 236

12.7 Basic mechanisms of antioxidant action 247

12.8 Interfacial phenomena – studies in heterogeneous lipid systems 252

12.9 Effect of temperature 265

Acknowledgments 267

References 267

13 Evaluation of antioxidant activity/capacity measurement methods for food products 273
Esra Capanoglu, Senem Kamiloglu, Gulay Ozkan, and Resat Apak

13.1 Introduction 273

13.2 Antioxidant assay selection for different food products 276

13.3 General conclusions and future perspectives 281

References 283

14 Antioxidants in oxidation control 287
Fereidoon Shahidi and Priyatharini Ambigaipalan

14.1 Introduction 287

14.2 Oxidation 287

14.3 Antioxidants 288

14.4 Synthetic antioxidants 289

14.5 Natural antioxidants 289

14.6 Tocols 290

14.7 Ascorbic acid 291

14.8 Carotenoids 292

14.9 Polyphenols 295

14.10 Bioavailability of phenolic antioxidants 307

14.11 Structural and other modification of phenolic antioxidants 308

14.12 Protein�]derived antioxidants 309

14.13 Phospholipids 309

14.14 Other antioxidants 310

References 310

15 Kinetic matching approach for rapid assessment of endpoint antioxidant capacity 321
Luis M. Magalhaes, Ines I. Ramos, Luisa Barreiros, Salette Reis, and Marcela A. Segundo

15.1 Introduction 321

15.2 Kinetic matching strategy 323

15.3 Expression of results as common standard 323

15.4 Application to samples 324

15.5 Conclusion 329

Acknowledgments 329

References 330

Index 333

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