**Author details**

vigorously by vortexing and perform an absorbance reading at 425 nm.

**Figure 2.** Standard quercetin curve (quercetin concentration × absorbance).

208 Honey Analysis

*Calculation of total flavonoid:* using absorbance values (*y*) and the linear equation find the *x* value corresponding to the total flavonoid in quercetin equivalent/L. Then, multiply the values by their respective dilutions and obtain the final QE values in mg/L. Using the total flavonoid quercetin equivalent/1000 mL of the main quercetin solution, calculate the corresponding values in 10 mL of the honey solution (containing 0.4 g of honey/mL). From these results, calculate the total flavonoid concentration in quercetin equivalent/100 g of honey. Calculate the mean and standard deviation and express the results as quercetin equivalent/100 g of honey ± deviation found.

Antioxidant activity is determined by the scavenging capacity of the free radical DPPH (2,2-diphenyl-1-picrylhydrazyl). The method involves reducing an alcoholic solution of purple DPPH radicals, which, upon receiving an electron or hydrogen radical, changes color from violet to yellow (diphenyl-picryl hydrazine),accompanied by a decrease in absorbance at the wavelength observed [54]. The greater or lesser capacity of the sample to reduce DPPH, or in other words to prevent oxidation, is evidenced by the percentage of DPPH remaining in the system [55]. This free radical, stable at room temperature, is reduced in the presence of an

*Preparation of the DPPH solution 0.06 mM*: weigh 0.0023 g of DPPH (molecular weight = 4.32 g/L)

*Preparation of the initial honey solution:* weigh 8 g of honey and transfer to a 10 mL volumetric flask using methanol as solvent, to obtain a solution with a honey concentration equal to 800 mg/mL of the main solution. From this concentration, obtain 1.0 mL of the diluted solutions with 80.0, 120.0, 200.0, 400.0, 600.0 and 800.0 mg of honey/mL. Calculate honey concen-

Honey (mg/mL) <sup>=</sup> (mg honey/mL main solution <sup>×</sup> pipetted volume (mL)) <sup>×</sup> <sup>100</sup> (17)

Volume of the main solution pipetted (mL) = 0.10, 0.15, 0.25, 0.50, 0.75 and 1.00 mL.

**2.16. Ability to kidnap stable free radical 2,2-diphenyl-1-picrylhydrazyl—DPPH**

antioxidant molecule, yielding a yellow solution.

and transfer to a 100 mL volumetric flask using methanol as solvent.

trations per mL (mg/mL) of diluted solutions applying Eq. (17):

Adjust the volume of solution to 1.0 mL using methanol as solvent.

where honey in the main solution (mg/mL) = 80 mg/mL.

Maria Josiane Sereia1 \*, Paulo Henrique Março1 , Marcia Regina Geraldo Perdoncini1 , Rejane Stubs Parpinelli2 , Erica Gomes de Lima<sup>2</sup> and Fernando Antônio Anjo3

\*Address all correspondence to: mjsereia@gmail.com

1 Department of Engineering and Food Technology, Federal Technology University of Parana, Campo Mourão, Brazil

2 Department of Animal Science, State University of Maringa, Maringa, Brazil

3 Agricultural Science Center, State University of Maringa, Maringa, Brazil
