Targeted and semi-untargeted determination of phenolic compounds in plant matrices by high performance liquid chromatography-tandem mass spectrometry

Abstract

In this work two different acquisition approaches were used for the quantification and/or tentative identification of phenolic compounds (PCs) in plant matrices by HPLC-MS/MS. A targeted approach, based on MRM acquisition mode, was used for the identification and quantification of a list of target analytes by comparison with standards; a semi-targeted approach was also developed by the precursor ion scan and neutral loss for the tentative identification of compounds not included in the target list.Analysis of phenolic content in three different plant matrices (curry leaves, hemp and blueberry) was carried out. The extraction and clean-up steps were set up according to the characteristics of the sample allowing to minimize the interfering compounds present in such complex matrices, as proved by the low matrix effect obtained (<16%) and recovery values ranging from 45% to 98% for all the analytes.This approach provided a sensitive and robust quantitative analysis of the target compounds with LOQs between 0.0002 and 0.05 ng mg−1, which allowed the identification and quantification of several hydroxycinnamic and hydroxybenzoic acids, in addition to numerous flavonoids in all three matrices.Furthermore, different moieties were considered as neutral losses or as precursor ions in semi-targeted MS/MS approach, providing the putative identification of different glycosylated forms of flavonoids, such as luteolin-galactoside and diosmin in all three matrices, while apigenin-glucuronide was detected in hemp and quercetin-glucuronide in blueberry.A further study was carried out by MS3, allowing the discrimination of compounds with similar aglycones, such as luteolin and kaempferol.