In vitro antioxidant activity evaluation of rosmaric acid and honey based supplement

Disrupted balance between free radicals and antioxidants leads to a pathophysiological state of oxidative stress, which is the underlying cause of many diseases. Since some synthetic antioxidants show some potentially adverse effects, many studies aim to find natural antioxidants. Secondary plant metabolites, such as polyphenols, phenolic acids, which include rosmarinic acid, terpenes and terpenoids have antioxidative properties and can be found in different hydrosols and honey. The aim of this study is to evaluate the in vitro antioxidative activity of the supplement based on hydrosol mixture, rosmarinic acid and honey. A series of diluted (100%, 50%, and 25%) supplements were added to the serum pool, collected from healthy donors. A second series of samples was prepared with the same amounts of supplement and addition of tert-Butyl hydroperoxide (TBH) as a prooxidant. Alongside these samples, sera samples with Trolox (hydrophilic vitamin E analog) and Trolox with TBH were prepared. All these samples were tested for the following oxidative stress parameters: total antioxidative status (TAS), paraoxonase-1 (PON-1), total sulfhydryl groups content (SHG), total oxidative status (TOS), prooxidative-antioxidative balance (PAB) and advanced oxidation protein products (AOPP). TAS/TOS ratio was calculated as a quantitative measurement of antioxidative and oxidative substances ratio in the serum. Antioxidative parameters TAS, SHG, TAS/TOS ratio and prooxidant/antioxidant balance parameter PAB, were not significantly different for the supplemened samples with and without the addition of TBH, which indicates that the supplement keeps its antioxidative properties, despite the addition of TBH. The supplement showed significant antioxidative properties as a result of the synergistic effect of rosmarinic acid, terpenes, and terpenoids from hydrosols and polyphenols and other antioxidant substances from honey.