Electrospray ionisation mass spectrometric behaviour of flavonoid 5-O-glucosides and their positional isomers detected in the extracts from the bark of Prunus cerasus L. and Prunus avium L.

Abstract

Literature data concerning the electrospray ionisation mass spectrometry (ESI-MS) behaviour of flavonoid 5-O-glycosides are poor and sometimes disputable. Therefore, we decided to analyse the compounds of this kind present in the bark of Prunus cerasus and Prunus avium by using high-performance liquid chromatography HPLC/ESI-MS. The aim of this study is to obtain the comprehensive information about the ESI-MS(+/-) behaviour of flavonoid 5-O-glucosides, to compare their behaviour with that of their positional isomers, to confirm that the known susceptibility of flavonoid 5-O-glucosides to hydrolysis may be successfully used for their identification. The bark from Prunus trees was extracted with pure methanol or, in order to perform hydrolysis and extraction simultaneously, with 5% methanolic solution of hydrochloric acid. The HPLC-ESI-MS analyses were performed using a Waters model 2690 HPLC pump and Waters/Micromass ZQ2000 mass spectrometer. Flavonoid 5-O-glycosides were completely hydrolysed under the acid conditions used, in contrast to their positional isomers. In positive ion mode, at low cone voltage, flavonoid 5-O-glycosides yield abundant Y0 + aglycone ions, in contrast to their positional isomers. In the negative ion mode, flavonoid 5-O-glycosides do not yield [Y0 - H]-· fragment ions, in contrast to their positional isomers. When aglycone contains only two hydroxyl groups, the flavonoid 5-O-glycosides can be detected in negative ion mode, whereas their positional isomers do not yield [M - H]- ions. It has been demonstrated that the susceptibility to hydrolysis of the analysed compounds, the abundances of respective fragment ions formed, and their ESI(-) response allow unambiguous identification of flavonoid 5-O-glycosides and their differentiation from their positional isomers.