Comparative evaluation of analytical methods for determining the antioxidant activities of honey: A review

Figures & data

Scheme 1. Reaction mechanisms of single-electron transfer (SET) and hydrogen atom transfer (HAT).

Figure 1. Mechanisms of antioxidant effects of honey (Al-mamary et al., 2002).

Figure 2. Total antioxidant activity of honey samples of different botanical origin (Aldina et al., 2015).

Table 1. Phenolic and Flavonoid content, and antioxidant activity of eleven Tualang honey samples (Siti et al., 2012)

Parameters	Mean
TPC (mg GAE/kg honey)	383.61 ± 34.53
FC (mg CEQ/kg honey)	149.77 ± 12.83
DPPH (%)	34.93 ± 2.44
FRAP (μM Fe(II)/kg	281.38 ± 5.99

Table 2. Correlation established between phenolic compounds and the antioxidant activity of Tualang honey samples (Siti et al., 2012)

Parameters	Equation	R^2	P
TPC
DPPH	Y = 0.0426X + 18.5713	0.334	<0.001
FRAP	Y = 0.0664X + 255.897	0.093	0.084
FC
DPPH	Y = 0.1356X + 14.4294	0.335	<0.001
FRAP	Y = 0.3238X + 232.444	0.219	0.006

Table 3. Correlation matrix showing the interrelation among phenolics, flavonoids, DPPH scavenging and FRAP assay for Malaysian honeys (Siti et al., 2013)

	TPC	FC	DPPH	FRAP
TPC	1
FC	0.989**	1
DPPH	0.804**	0.848**	1
FRAP	0.914**	0.848**	0.712**	1

Figure 3. Correlation between polyphenol content and antioxidant activity of honey (Aldina et al., 2015).

Table 4. Correlation matrix between antioxidant activity and biochemical components of honey (Lee et al., 2013)

	TPC	FC	DPPH	FRAP
TPC	1
FC	0.8375*	1
DPPH	0.5728*	0.9276	1
FRAP	0.9033	0.9910*	0.8691	1

Table 5. Pearson correlation matrixes for AAE-DPPH, TP, Color and TF (Natacha et al., 2014)

		AAE-DPPH*	TP	Colour	TF	AAE-RedPW
AAE-DPPH*	Correlation
	P-value
TP	Correlation	0.605
	P-value	0.000
Colour	Correlation	0.622	0.808
	P-value	0.000	0.000
TF	Correlation	0.062	0.008	−0.147
	P-value	0.563	0.937	0.166
AAE-RedPW	Correlation	0.597	0.894	0.830	−0.001
	P-value	0.000	0.000	0.000	0.996
EC50- RedPW	Correlation	−0.436	−0.703	−0.554	−0.022	−0.696
	P-value	0.000	0.000	0.000	0.836	0.000

Values in bold are the significant pearson correlation coefficients (P < 0.05)

Table 6. Correlations between total phenolic contents and RSA of honey samples (Mekuanint & Meareg, 2019)

	TPC	AEAC	RSA	IC50
TPC	1
AEAC	0.938**	1
RSA	0.940**	1.000**	1
IC50	− 0.884**	− 0.968**	− 0.970**	1

Table 7. Comparison of the analytical methods used for assessing the antioxidant activity (Ronald et al., 2005)

Antioxidant assay	Simplicity	Instrumentation required	Biological relevance	Mechanism	Lipophylic & hydrophilic AOC
ORAC	+	_	+++	HAT	+++
FRAP	++	+++	_ _	SET	_ _ _
TEAC	+	+	_	SET	+++
DPPH	+++	+++	_	SET/HAT	_

+, ++, +++ = desirable to highly desired characteristic; −, −−, −−− = less desirable to highly undesirable based upon this characteristic.

Scheme 2. Chemical structure of 2, 2-diphenyl-1-picrylhydrazyl (DPPH•).

Scheme 3. Conversion of Fe³⁺ TPTZ to Fe²⁺ TPTZ in FRAP assay.

Scheme 4. Reaction mechanisms in ORAC method.

Scheme 5. Formation of peroxyl radical (Joseph, 2004).

Figure 4. ORAC antioxidant activity expressed as the net area under the curve (Alberto, Carmen, & Begoña, 2004).

Figure 5. Kinetics of fluorescein (FL) quenching with different concentrations of Phycocyanobilin (PCB) versus blank. The reaction mixture contains 0.05 µM FL and 4 µM AAPH, with PCB ranging from 0.025 to 0.150 µM. (Inset) Linear plot of the net AUC versus PCB concentrations (Serena et al., 2010).

Figure 6. Time course of the reaction of fluorescein (FL) and β-phycoerythrin in the presence and in the absence of AAPH with and without antioxidant (wine) (Huang et al., 2002).

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