Determination of Callistephus chinensis flower petals anthocyanins by reverse-phase HPLC

Abstract

The specific composition of anthocyanin types of flower petals of Callistephus chinensis several varieties is determined by the inverse-phase HPLC method with mass-spectrometric and diode array detection. It was found that in the petals of all the samples under investigation anthocyanins were synthesized upon non-methylated aglycones, delphinidin, cyanidin and pelargonidin, proving the absence in flower cells methyl transferase. It was also established that two types of anthocyanin biosynthesis may result in accumulation of either 3-glucosides (3G) or 3,5-diglucosides (3,3diG) of some of above mentioned aglycones depending upon flower cultivars, with high degree of acylation by malonic acid. This results are in full agreement wiyh known literature data. The main acylation products (in position 6 of 3-glucosidic substituents) was accomplished by small quantities of isomers being malonated in position 2 of 4 of sugar substituent, but not at 3 position. All main possible types of anthocyanins may be separated by proposed method, but one cultivar with more complex anthocyanin composition was also found. In this case substantial quantities of 3G simultaneously with 3,5diG type of anthocyanins were present. Hence new type of inheritance of characters was received by the breeders. Results of retention analysis indicated that retention of anthocyanins after acylation by malonic acid (in logarithms units of retention factor) depends mainly upon the type of glycosylation and position of acylation rather than upon aglycone type and upon mobile phase composition. Meanwhile the retention of anthocyanins is a complex function of aglycon and the sugar moieties structures.