Extraction and Quantification of Abscisic Acid and Derivatives in Strawberry by LC-MS
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Extraction and Quantification of Abscisic Acid and Derivatives in Strawberry by LC-MS Ellen Cristina Perin 1 & Rosane Lopes Crizel 1 & Vanessa Galli 2 & Rafael da Silva Messias 1 & Cesar Valmor Rombaldi 1 & Fabio Clasen Chaves 1 Received: 29 December 2017 / Accepted: 1 March 2018 # Springer Science+Business Media, LLC, part of Springer Nature 2018
Abstract Phytohormones are important plant components that are involved in a signaling cascade in plant development. In strawberry, the influence of abscisic acid (ABA) and its different forms [phaseic acid (PA), dihydrophaseic acid (DPA), and ABA glucose ester (ABA-GE)] on the process of fruit maturation is not yet completely understood. Quantification of phytohormones is currently performed by liquid chromatography coupled to mass spectrometry (LC-MS) due to its sensitivity and specificity. However, the sample matrix and the extraction procedure will influence the analysis. Thus, this study aimed to optimize a simple extraction method and validate the LC-MS quantification of ABA as well as the identification and quantification of ABA derivatives (PA, DPA, and ABA-GE) in strawberry fruit. Hormone extraction was performed using either methanol (80% v/v—S1) or acetone:water:acetic acid (80:19:1 v/v—S2) solutions with or without the use of sonication. The most efficient extraction was obtained using S1 without sonication, and LC-MS validation parameters for ABA were within acceptable scores. Keywords Fragaria × ananassa . Abscisic acid . Abscisic acid glucose ester . Phaseic acid . Dihydrophaseic acid . Liquid chromatography-mass spectrometry
Introduction Strawberry (Fragaria × ananassa Duch.) pseudo fruit, henceforth called fruit, has sensory, nutritional, and functional properties recognized and appreciated by consumers. The ripening process influences the sensory and functional characteristics of strawberry fruit. Although the reactions involved in strawberry maturation are still not fully characterized, it has been suggested that abscisic acid (ABA) is involved in this process as a regulator (Ayub et al. 2016). ABA also plays a role in osmotic stress response by acting in stomatal closure as well as induction of the phenylpropanoid metabolic pathway and accumulation of antioxidant compounds (Chen et al. 2016a, b; Li et al. 2015). ABA levels in plants are regulated by ABA biosynthesis and catabolism pathways. The first step exclusively * Fabio Clasen Chaves [email protected] 1
Programa de Pós-graduação em Ciência e Tecnologia de Alimentos, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brazil
2
Biotecnologia, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brazil
committed to ABA biosynthesis is the conversion of the carotenoid zeaxanthin to trans-violaxanthin catalyzed by zeaxanthin epoxidase. Subsequently, a 9-cis-epoxycarotenoid dioxygenase catalyzes the oxidative cleavage of 9-cisvioloxanthin and/or 9-cis-neoxanthin producing xanthoxin, a rate-limiting step in ABA biosynthesis. Xanthoxin is eventually exported to the cytosol a
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