Rapid method for quantification of anthocyanidins and anthocyanins in human biological samples

Abstract

Using simple solvent extraction and enzymatic hydrolysis, a rapid LC-MS/MS method for quantification of free and conjugated forms of anthocyanidins and anthocyanins in plasma and urine samples was developed and validated. A mixed enzymatic treatment containing β-glucuronidase (100 U mL^-1) and sulfatase (2.5 U mL^-1) for 5 min (37 °C; pH 6) was optimal condition for deconjugation of anthocyanidins and anthocyanins in urine and plasma samples. The LC-MS/MS allowed quantifying thirteen different anthocyanidins and anthocyanins simultaneously. The developed LC-MS/MS method was precise and accurate over multiple days and nominal concentrations. The stability assessment study confirmed that the long-term storage and/or periodic use of plasma and urine samples might have a considerable impact on the stability of some anthocyanidins. The method was successfully applied to measure anthocyanidins and anthocyanins in plasma and urine samples following consumption of acute blueberry test meals.

Keywords: Acute blueberry consumption; Anthocyanidin; Anthocyanin; Plasma; Urine.