Investigation of contribution of individual constituents to antioxidant activity in herbal drugs using postcolumn HPLC method

The most important attention is paid to the search of natural antioxidants and their evaluation in medicinal and food raw materials of plant origin. A number of plants, their extracts, food products, and medicinal preparations appear to be the objects of scientific research. Effectiveness and informative character of research, undoubtedly, depend on relevance, sensitivity, and efficiency of the methods chosen. The aim of this work was to develop and validate the postcolumn high-performance liquid chromatography (HPLC)-DPPH method as well as its application in the evaluation of antioxidant activity of known and unknown compounds scavenging free radicals and existing in medicinal plant raw materials. HPLC-separated compounds were identified at the wavelength of 275 nm, and then the mobile phase with analytes flowed through a mixing tee to the reaction coil, where DPPH reagent solution was supplied. The solution flow rate was 0.4 mL/min. The reaction coil was connected with UV/VIS type detector, which measured absorption of flowing solution at the wavelength of 520 nm. It was determined that vitexin rhamnoside, the dominant compound in the leaves of Crataegus monogyna, was not a significant radical scavenger. The most active antioxidant in the leaves and flowers of Crataegus monogyna was chlorogenic acid. The most active antioxidant in Origanum vulgare raw material was rosmarinic acid. Identified analytes in the extracts of Achillea millefolium that possessed radical-scavenging properties were chlorogenic acid, luteolin-7-Oglucoside, rutin, and luteolin.