Chapter 9
Analytical Protocols in Antioxidant
Capacity Measurement
Tatiele Casagrande do Nascimento, Renan Campos Chisté,
and Leila Queiroz Zepka
Abstract Microalgae have been recognized as promising sources of carotenoids,
chlorophylls, phycobiliproteins, and phenolic compounds, which are bioactive
compounds widely investigated in the decrease of oxidative damage. The ability
of these structures to scavenge reactive chemical species occurs by different mechanisms and can be determined by several assays. Among the existing protocols, there
are non-competitive [DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid), FRAP (Ferric Reducing Antioxidant
Power), and RC (Reducing Capacity by Folin-Ciocalteu assay)] and competitive
methods [ORAC-H (Oxygen Radical Absorbance Capacity for hydrophilic antioxidants), ORAC-L (Oxygen Radical Absorbance Capacity for lipophilic antioxidants)
and PRSC (Peroxyl Radical Scavenging Capacity)]. Each protocol has a specific reaction medium, which can lead to some limitations, and therefore, its use cannot be
generalized. Considering that the microalgae biomass has compounds with different
structural characteristics for the solubility (hydrophilic and lipophilic), the compatibility of the reaction medium with the structure must be considered for an unequivocal measurement. Based on this context, this chapter presents a comprehensive
description of the most used protocols to determine the in vitro antioxidant capacity
of microalgae compounds, including the structural characteristics of the bioactive
compounds, reaction medium, principles, mechanisms, and limitations of existing
protocols, as well as a database of existing measurements.
Keywords Microalgae · Bioactive compounds · Antioxidant capacity
T. Casagrande do Nascimento · L. Queiroz Zepka
Department of Food Science and Technology, Federal University of Santa Maria (UFSM),
97105-900 Santa Maria, RS, Brazil
e-mail: tatielecasagrande@gmail.com
L. Queiroz Zepka
e-mail: zepkaleila@yahoo.com.br
R. Campos Chisté (B)
Faculty of Food Engineering (FEA), Institute of Technology (ITEC), Federal University of Pará
(UFPA), 66075-110 Belém, PA, Brazil
e-mail: rcchiste@ufpa.br
© Springer Nature Switzerland AG 2020
E. Jacob-Lopes et al. (eds.), Pigments from Microalgae Handbook,
https://doi.org/10.1007/978-3-030-50971-2_9
203
Analytical Protocols in Antioxidant
Capacity Measurement
Tatiele Casagrande do Nascimento, Renan Campos Chisté,
and Leila Queiroz Zepka
Abstract Microalgae have been recognized as promising sources of carotenoids,
chlorophylls, phycobiliproteins, and phenolic compounds, which are bioactive
compounds widely investigated in the decrease of oxidative damage. The ability
of these structures to scavenge reactive chemical species occurs by different mechanisms and can be determined by several assays. Among the existing protocols, there
are non-competitive [DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid), FRAP (Ferric Reducing Antioxidant
Power), and RC (Reducing Capacity by Folin-Ciocalteu assay)] and competitive
methods [ORAC-H (Oxygen Radical Absorbance Capacity for hydrophilic antioxidants), ORAC-L (Oxygen Radical Absorbance Capacity for lipophilic antioxidants)
and PRSC (Peroxyl Radical Scavenging Capacity)]. Each protocol has a specific reaction medium, which can lead to some limitations, and therefore, its use cannot be
generalized. Considering that the microalgae biomass has compounds with different
structural characteristics for the solubility (hydrophilic and lipophilic), the compatibility of the reaction medium with the structure must be considered for an unequivocal measurement. Based on this context, this chapter presents a comprehensive
description of the most used protocols to determine the in vitro antioxidant capacity
of microalgae compounds, including the structural characteristics of the bioactive
compounds, reaction medium, principles, mechanisms, and limitations of existing
protocols, as well as a database of existing measurements.
Keywords Microalgae · Bioactive compounds · Antioxidant capacity
T. Casagrande do Nascimento · L. Queiroz Zepka
Department of Food Science and Technology, Federal University of Santa Maria (UFSM),
97105-900 Santa Maria, RS, Brazil
e-mail: tatielecasagrande@gmail.com
L. Queiroz Zepka
e-mail: zepkaleila@yahoo.com.br
R. Campos Chisté (B)
Faculty of Food Engineering (FEA), Institute of Technology (ITEC), Federal University of Pará
(UFPA), 66075-110 Belém, PA, Brazil
e-mail: rcchiste@ufpa.br
© Springer Nature Switzerland AG 2020
E. Jacob-Lopes et al. (eds.), Pigments from Microalgae Handbook,
https://doi.org/10.1007/978-3-030-50971-2_9
203