High-performance liquid chromatography with post-column 2,2′-diphenyl-1-picrylhydrazyl radical scavenging assay: Methodological considerations and application to complex samples Original Research Article

An improved post-column 2,2′-diphenyl-1-picrylhydrazyl (DPPHradical dot) radical scavenging assay for the screening of antioxidants in complex matrices was developed. Experimental parameters believed to be influential to DPPHradical dot response were studied in a univariate approach. Optimum conditions were found to be: 5 × 10−5 M DPPHradical dot reagent prepared in a 75% methanol: 25% 40 mM citric acid–sodium citrate buffer (pH 6) solution, degassed with nitrogen; reaction coil of 2 m × 0.25 mm i.d. PEEK tubing; detection at 521 nm; analysis at room temperature. The analytical utility of this protocol was evaluated by screening for antioxidants in thyme and green tea, in comparison with two commonly employed methodologies.