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Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis

Soluble sugars play an important role in freezing tolerance in both herbaceous and woody plants, functioning in both the reduction of freezing-induced dehydration and the cryoprotection of cellular constituents. The quantification of soluble sugars in plant tissues is, therefore, essential in unders...

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Autores principales: Zhao, Lu, Chanon, Ann M., Chattopadhyay, Nabanita, Dami, Imed E., Blakeslee, Joshua J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904305/
https://www.ncbi.nlm.nih.gov/pubmed/27379118
http://dx.doi.org/10.3389/fpls.2016.00818
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author Zhao, Lu
Chanon, Ann M.
Chattopadhyay, Nabanita
Dami, Imed E.
Blakeslee, Joshua J.
author_facet Zhao, Lu
Chanon, Ann M.
Chattopadhyay, Nabanita
Dami, Imed E.
Blakeslee, Joshua J.
author_sort Zhao, Lu
collection PubMed
description Soluble sugars play an important role in freezing tolerance in both herbaceous and woody plants, functioning in both the reduction of freezing-induced dehydration and the cryoprotection of cellular constituents. The quantification of soluble sugars in plant tissues is, therefore, essential in understanding freezing tolerance. While a number of analytical techniques and methods have been used to quantify sugars, most of these are expensive and time-consuming due to complex sample preparation procedures which require the derivatization of the carbohydrates being analyzed. Analysis of soluble sugars using capillary zone electrophoresis (CZE) under alkaline conditions with direct UV detection has previously been used to quantify simple sugars in fruit juices. However, it was unclear whether CZE-based methods could be successfully used to quantify the broader range of sugars present in complex plant extracts. Here, we present the development of an optimized CZE method capable of separating and quantifying mono-, di-, and tri-saccharides isolated from plant tissues. This optimized CZE method employs a column electrolyte buffer containing 130 mM NaOH, pH 13.0, creating a current of 185 μA when a separation voltage of 10 kV is employed. The optimized CZE method provides limits-of-detection (an average of 1.5 ng/μL) for individual carbohydrates comparable or superior to those obtained using gas chromatography–mass spectrometry, and allows resolution of non-structural sugars and cell wall components (structural sugars). The optimized CZE method was successfully used to quantify sugars from grape leaves and buds, and is a robust tool for the quantification of plant sugars found in vegetative and woody tissues. The increased analytical efficiency of this CZE method makes it ideal for use in high-throughput metabolomics studies designed to quantify plant sugars.
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spelling pubmed-49043052016-07-04 Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis Zhao, Lu Chanon, Ann M. Chattopadhyay, Nabanita Dami, Imed E. Blakeslee, Joshua J. Front Plant Sci Plant Science Soluble sugars play an important role in freezing tolerance in both herbaceous and woody plants, functioning in both the reduction of freezing-induced dehydration and the cryoprotection of cellular constituents. The quantification of soluble sugars in plant tissues is, therefore, essential in understanding freezing tolerance. While a number of analytical techniques and methods have been used to quantify sugars, most of these are expensive and time-consuming due to complex sample preparation procedures which require the derivatization of the carbohydrates being analyzed. Analysis of soluble sugars using capillary zone electrophoresis (CZE) under alkaline conditions with direct UV detection has previously been used to quantify simple sugars in fruit juices. However, it was unclear whether CZE-based methods could be successfully used to quantify the broader range of sugars present in complex plant extracts. Here, we present the development of an optimized CZE method capable of separating and quantifying mono-, di-, and tri-saccharides isolated from plant tissues. This optimized CZE method employs a column electrolyte buffer containing 130 mM NaOH, pH 13.0, creating a current of 185 μA when a separation voltage of 10 kV is employed. The optimized CZE method provides limits-of-detection (an average of 1.5 ng/μL) for individual carbohydrates comparable or superior to those obtained using gas chromatography–mass spectrometry, and allows resolution of non-structural sugars and cell wall components (structural sugars). The optimized CZE method was successfully used to quantify sugars from grape leaves and buds, and is a robust tool for the quantification of plant sugars found in vegetative and woody tissues. The increased analytical efficiency of this CZE method makes it ideal for use in high-throughput metabolomics studies designed to quantify plant sugars. Frontiers Media S.A. 2016-06-13 /pmc/articles/PMC4904305/ /pubmed/27379118 http://dx.doi.org/10.3389/fpls.2016.00818 Text en Copyright © 2016 Zhao, Chanon, Chattopadhyay, Dami and Blakeslee. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Zhao, Lu
Chanon, Ann M.
Chattopadhyay, Nabanita
Dami, Imed E.
Blakeslee, Joshua J.
Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis
title Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis
title_full Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis
title_fullStr Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis
title_full_unstemmed Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis
title_short Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis
title_sort quantification of carbohydrates in grape tissues using capillary zone electrophoresis
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904305/
https://www.ncbi.nlm.nih.gov/pubmed/27379118
http://dx.doi.org/10.3389/fpls.2016.00818
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