Anthocyanins, Carotenoids and Chlorophylls in Edible Plant Leaves Unveiled by Tandem Mass Spectrometry

Natural pigments are a quite relevant group of molecules that are widely distributed in nature, possessing a significant role in our daily lives. Besides their colors, natural pigments are currently recognized as having relevant biological properties associated with health benefits, such as anti-tumor, anti-atherogenicity, anti-aging and anti-inflammatory activities, among others. Some of these compounds are easily associated with specific fruits (such as blueberries with anthocyanins, red pitaya with betalain or tomato with lycopene), vegetables (carrots with carotenoids), plant leaves (chlorophylls in green leaves or carotenoids in yellow and red autumn leaves) and even the muscle tissue of vertebrates (such as myoglobin). Despite being less popular as natural pigment sources, edible plant leaves possess a high variety of chlorophylls, as well as a high variety of carotenoids and anthocyanins. The purpose of this review is to critically analyze the whole workflow employed to identify and quantify the most common natural pigments (anthocyanin, carotenoids and chlorophylls) in edible plant leaves using tandem mass spectrometry. Across the literature there, is a lack of consistency in the methods used to extract and analyze these compounds, and this review aims to surpass this issue. Additionally, mass spectrometry has stood out in the context of metabolomics, currently being a widely employed technique in this field. For the three pigments classes, the following steps will be scrutinized: (i) sample pre-preparation, including the solvents and extraction conditions; (ii) details of the chromatographic separation and mass spectrometry experiments (iii) pigment identification and quantification.