This study presents the optimization and validation of a rapid protocol for quantifying alkyresorcinol (AR) metabolites 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA) in plasma, using high-performance liquid chromatography (HPLC) coupled with a coulometric electrode array detector. Syringic acid (SyrA) serves as the internal standard. The new method is simple and could be used in large epidemiological studies. The summed AR metabolite concentrations measured in plasma correlate significantly with the summed urinary AR metabolite concentrations (R = 0.613; p < 0.001) and with the summed intact AR (C17:0-C25:0) concentrations in plasma (R = 0.686; p < 0.001). Additional investigation is needed to clarify whether the two plasma AR metabolites are useful as biomarkers of whole-grain intake and helpful in the exploration of the association between whole-grain cereal intake and human diseases.