Validation of HPLC assay for the identification and quantification of anthocyanins in black currants
Many publications describe separation and quantification techniques for anthocyanins. However, chromatographic conditions to optimize the separation were seldom reported. We developed a sensitive and reliable HPLC method and validated the method for anthocyanin separation, identification, and quantification. Separation was performed on a Zorbax SB C18 column. The conditions including ion pair reagent, mobile phase pH, and mobile phase flow rate were optimized to improve the separation efficiency. Tandem MS was used to identify the total 6 anthocyanins in black currant. Validation of the developed method showed good linearity, precision, repeatability, and excellent recovery. The validated method was used to quantify anthocyanins from black currant samples.