A new acetonitrile-free mobile phase for HPLC-DAD determination of individual anthocyanins in blackcurrant and strawberry fruits: A comparison and validation study.

Research focused on identification and quantification of anthocyanins from berries and other fruits is gaining importance due to the observed inverse relationship between anthocyanin intake and the incidence of certain diseases. Separation and quantification of these compounds is mainly achieved on reverse phase HPLC coupled to different detection systems using mostly acetonitrile as the mobile phase of choice. Nevertheless, the scientific community recently faced a worldwide shortage of this solvent which resulted in prices soaring dramatically. In this context, the present study describes the comparison and validation of a newly developed methanol-based method for the identification and quantification of major berry anthocyanins using standard HPLC coupled to photo diode array detection. Moreover, two different commercially available stationary phases were tested. The methanol-based method developed herein showed high repeatability (R.S.D <1.3%), rapidity (<35min) and accuracy and therefore may be suitable for routine quantification of berry anthocyanins. Comparison with an earlier established acetonitrile-based method showed that despite differences in absolute concentrations between both methods the determined anthocyanin concentrations were highly correlated (r>0.95). Method validation was further achieved by elucidating differences in the anthocyanin profile between different blackcurrant and strawberry cultivars.