Simple and Fast Method for Identification and Quantification of Anthocyanidins in Berries by Ultra Performance Liquid Chromatography-Mass Spectrometry

Abstract

The objective was identifying three different anthocyanidins and to quantify them in six different berries. Results from the conventional method (using extraction with hydrochloric acid) were compared with data from the proposed method (without acid extraction). The following figures of merit were used: linearity, accuracy, linear range, limits of detection and quantification, using an ultra performance liquid chromatography (UPLC) coupled to a triple quadrupole mass spectrometer equipped with an electrospray ionization source. The separation of anthocyanidins can be achieved in a much smaller period of time (1.30 min) and the cyanidin from Morus nigra L., Rubus idaeus L. and Vaccinium myrtillus L. was found in higher amounts in extracts obtained from the proposed method. Proposed mechanisms of mass spectrometry (MS/MS) collisional induced dissociation for pelargonidin, cyanidin and delphinidin were obtained and this is the first time that concentrations of anthocyanidins present in fruits of Solanum americanum Mill. were reported.

Anthocyanidins structural study using positive electrospray ionization triple quadrupole mass spectrometry and H/D exchange

“…The fragmentation study of the anthocyanins and their deuterated species were fundamental to propose reliable mechanism; the general mechanism proposed in a previous work in an analytical method development was revisited and new fragmentation channels proposed. Isotope labeling experiments were performed for all anthocyanins preparing solutions with methanol‐d4 and waiting the H/D exchanges to directly analyze them by ESI‐MS and ESI‐MS/MS.…”

“…Since the water loss is thermodynamically less favored than CO loss, the subsequent fragmentation proposal was focused on the 3′ ion of m/z 243, which is a result from CO loss from position C3 toward carbon monoxide elimination with charge retention fragmentation (CRF), which did not break the aromaticity. It did not discard other more energetic possibilities with subsequent H 2 O and/or CO losses, but the thermodynamically more favorable fragmentation has been followed.…”

“…The use of triple quadrupole mass spectrometer in fragmentation experiments provides the confidence to propose fragmentation pathways together to ab initio calculations. 17 The fragmentation study of the anthocyanins and their deuterated species were fundamental to propose reliable mechanism; the general mechanism proposed in a previous work 18 Since the water loss is thermodynamically less favored than CO loss, the subsequent fragmentation proposal was focused on the 3′ ion of m/z 243, which is a result from CO loss from position C3 toward carbon monoxide elimination with charge retention fragmentation (CRF), 18 which did not break the aromaticity. It did not discard other more energetic possibilities with subsequent H 2 O and/or CO losses, but the thermodynamically more favorable fragmentation has been followed.…”

Quantification of anthocyanosides in grapes by QuEChERS and biphenyl-UHPLC tandem mass spectrometry

“…The octadecyl-silica (ODS, C18) column is widely used for the separation of individual AnCS derivatives for various fruits [ 17 ]. However, despite the well-documented benefits of the C18 stationary phase for the determination of some types of AnCS [ 18 ], the similar structures of the glycosylated AnCS derivatives combined with their structural diversity complicate their analysis, and the complete separation of natural mixtures of complex AnCS is not always possible. In the recent development of separation targeted for liquid chromatography, the use of a pentafluorophenyl (F5) stationary phase has received considerable attention.…”