High Specificity and Sensitivity Method for Quantitation of Anthocyanins by Precursor Ion Scan Using Liquid Chromatography-Tandem Mass Spectrometry Technique

Anthocyanins, primarily found in flowers, vegetables, and fruits, play a critical role in human and plant health. These polyphenolic pigments are predominantly found in plants in the form of glucosides linked to one or more sugar units and exhibit a broad range of structural diversity. This creates a complex spectrum of anthocyanins making it challenging to perform its absolute quantification in different plant matrices due to the lack of availability of all reference standards. More commonly today, anthocyanins are quantified by spectrophotometric methods, which is non-specific quantification due to a lack of information on the anthocyanin classes present. In this study, a systematic approach has been proposed to address this challenge, utilizing liquid chromatography-tandem mass spectrometry using precursor ion scan mode for high-specificity identification and subsequent quantification of the anthocyanin classes. A method was developed for six anthocyanin classes and it was used to quantify anthocyanins in black carrot extract. The initial scan was conducted for all anthocyanin classes, followed by the quantification of the anthocyanin classes detected using the reference standard specific to that class. This approach has demonstrated that precursor ion scan mass spectrometry can serve as a valuable tool for the rapid identification and specific quantification of anthocyanin classes in plants.