Separation and Characterization of Anthocyanins by HPLC

F1.3 Separation and Characterization of Anthocyanins by HPLC FI. 4 Characterization of Anthocyanins by NMR... 

Better separations of complex anthocyanin mixtures can be expected through the use of micro-HPLC and newly developed stationary phases. The quantification of individual anthocyanins will be improved by an increased application of pure anthocyanin standards. By direct combination of HPLC with MS, a new avenue in online characterization and identification of anthocyanins is opened, albeit limited by the high costs (201). 

C18 solid-phase extraction is used to fractionate polyphenolics for their identification and characterization. This technique can eliminate interfering chemicals from crude extracts and produce desirable results for HPLC or other analytical procedures. To obtain a sufficient volume for all analyses, several separations by solid-phase extraction may be performed. The individual fractions need to be combined and dissolved in solvents appropriate for HPLC analysis. In Basic Protocol 2, the application of a current of nitrogen gas for the removal of water from the C18 cartridge is an important step in the selective fractionation of polyphenolics into non-anthocy-anin and anthocyanin fractions. After the collection of non-anthocyanin polyphenolics, no additional work is necessary to elute anthocyanins bound to the C18 solid phase if anthocyanins are not to be determined. 

---