Approaches to the determination of antioxidant activity of extracts from bee bread and safflower leaves and flowers

The main objective of this study was to develop approaches for the determination of total antioxidant activity of natural products (bee bread and safflower extracts) using DPPH radical scavenging assay. Considering that analytical procedures and results related to this assay and reported by many authors are significally differed between each other and depend on many factors (the nature of tested extracts, the nature of solvents for extraction, a reaction time of DPPH with a sample, DPPH solvents and concentration, ratio between DPPH and an extract, etc.), the methodology of the evaluation of antioxidant capacity of different origin extracts by DPPH radical scavenging assay was developed. Ascorbic acid (AA) was used as standard antioxidant and the correlation between the percentage of DPPH scavenging and AA concentration was determined at two different initial absorbances of DPPH solution. Average concentration of AA which inhibited 50% of DPPH radicals (IC50) was equal to 156.0 - 171.26 µg.mL-1. The reaction kinetics of DPPH inhibition by bee bread and safflower extracts was described by the curves of the dependence of the total antioxidant activity on time with squared correlation coefficients (R2) in the range of 0.89 - 0.98. The reaction times for these extracts were from 40 to 70 min at the correct ratio of volumes between the tested extracts and a DPPH solution. These studies demonstrated that the extracts obtained from bee bread of 2016 year of pollen collection had significantly higher the total antioxidant activity compared with the extracts of bee bread of pollen collection of 2015 considering the ratio of bee bread and the solvent in the extracts and volume of the extract for the procedure. This fact is explained not only botanical origin bun also the time of the storage of bee bread before the preparation of extracts. There was not found significant differences in the total antioxidant activity of extracts from flowers of safflower sown in fall and in spring. Antioxidant activity of the extracts from leaves of spring sown safflower was higher compared with the total antioxidant activity of the extracts from fall sown plants. Ascorbic acid equivalents of the tested extracts could be ranged as follows: bee bread of 2016 pollen collection >bee bread of 2015 pollen collection >leaves of safflower spring sown >flowers of safflower spring sown >flowers of safflower fall sown >leaves of safflower fall sown.&nbsp