High Performance Liquid Chromatographic Identification of Anthocyanins

Abstract
A high performance liquid chromatographic procedure for a rapid identification of antho- cyanins is described. Besides their characteristic retention times, anthocyanins can easily be identified by an HPLC examination of the kinetics of appearance and disappearance of their partial hydrolysis products. This method is also applicable to 3-deoxyanthocyanins which is demonstrated with an extract from the fern Blechnum brasiliense. The described method allows optimal resolution of complex anthocyanin mixtures in the fol­lowing sequence of elution: 3-sophoroside-5-glucoside; 3-rutinoside-5-glucoside; 3,5-diglucoside; 3-sophoroside; 3-galactoside; 3-glucoside; 3-rutinoside; 5-glucoside; aglycone, and finally the forms acylated with p-coumaric acid. Substitution in the B-ring of the aglycone structure is the key factor for the range of the retention time. Thus cyanidin 3,5-diglucoside elutes earlier (-38 sec) than pelargonidin 3-sophoroside-5-glucoside.

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