Abstract
The influence of exchange of HCOOH (System 2) by phosphoric acid (System 1) for acidification of the “acetonitrile–water” mobile phases for reversed-phase HPLC of anthocyanins was investigated in the framework of relative retention analysis. The differences and similarities of anthocyanins separation were revealed. It has been shown that some common features of the quantitative relationships may be used for preliminary anthocyanins structure differentiation, according to the number of OH-groups in anthocyanidin backbone as well as to a number of saccharide molecules in glycoside radicals in position 3 of the anthocyanin without MS detection.
Highlights
Anthocyanins are powerful water-soluble antioxidants of flavonoids class with health promoting effect [1, 2]
Anthocyanins are glycosides of anthocyanidins (Figure 1), with great varieties of more than 600 anthocyanin structures found in plant sources [4] though only six structures of the latter cover the majority of the structures due to glycosylation type variability [5]
The aim of the present paper is the investigation of regularities of anthocyanins retention in RP HPLC with mobile phases composed of water, acetonitrile and phosphoric acid and comparing that with retention in the more commonly used mobile phases being water–acetonitrile mixtures acidified with HCOOH (10 vol %)
Summary
Anthocyanins are powerful water-soluble antioxidants of flavonoids class with health promoting effect [1, 2]. The coloured flavylium form of anthocyanins is a reason to regard them as natural food colorants [2]. The latter explains a high scientific and technological interest to the substances. Anthocyanins are synthesized in plant tissue, mainly in fruits, flowers and for some species in leaves as a rule as a complex mixture of compounds with different structures [3]. Reversed-phase HPLC is a common method for analysis of complex mixtures of plant anthocyanins [6, 7]. Acidification is necessary to transfer the substances into charged and coloured flavylium form, due to the fact that anthocyanins may be detected at the presence of large amounts of other colourless substances
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