Importance of Extract Standardization and In Vitro/Ex Vivo Assay Selection for the Evaluation of Antioxidant Activity of Botanicals: A Case Study on Three Rosmarinus officinalis L. Extracts

The overproduction of free radicals and oxygen reactive species is suspected to be implicated in a wide range of metabolic reactions that can have pernicious consequences in the development of a variety of human diseases. Botanical extracts are sources of antioxidants that counteract both free radicals and oxygen reactive species. The processing conditions used in the botanical extraction may influence the antioxidant composition; therefore, different extracts from the same plant may have different antioxidant properties. To illustrate this fact, we conducted a study using three commercial rosemary (Rosmarinus officinalis L.) leaf extracts. The three extracts were standardized to contain, respectively, 20% carnosic acid, 40% ursolic acid, or 20% rosmarinic acid. They were evaluated for their total (hydrophilic + lipophilic) antioxidant effects on oxygen radical absorbance capacity (ORAC), their ferric reducing/antioxidant power (FRAP), and their capacity to inhibit Cu(2+)-induced low-density lipoprotein (LDL) oxidation ex vivo. The ursolic acid extract showed the lowest antioxidant capacity on all models. The rosmarinic acid extract had an antioxidant capacity 1.5 times higher on ORAC and four times higher on FRAP than the carnosic acid extract. However, the carnosic acid extract was better than the rosmarinic acid extract in inhibiting the oxidation of LDL ex vivo. These results encourage conducting further studies to evaluate the carnosic acid and rosmarinic acid extracts in vivo. Our study offers an example of the importance of the extraction procedures, on which depends the nature of the antioxidant composition, and highlights interest to proceed with in vitro/ex vivo assay selection for the evaluation of the antioxidant properties of botanical extracts.