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Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)

High-performance liquid chromatography (HPLC) analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and qua...

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Detalles Bibliográficos
Autores principales: Bai, Cheng, Reilly, Charles C., Wood, Bruce W.
Formato: Texto
Lenguaje:English
Publicado: Libertas Academica 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2716812/
https://www.ncbi.nlm.nih.gov/pubmed/19662174
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author Bai, Cheng
Reilly, Charles C.
Wood, Bruce W.
author_facet Bai, Cheng
Reilly, Charles C.
Wood, Bruce W.
author_sort Bai, Cheng
collection PubMed
description High-performance liquid chromatography (HPLC) analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates). Asparagine alone is eluted earlier than citrulline alone, but when both of them are present in biological samples they may co-elute. HPLC retention times for asparagine and citrulline were influenced by other ureides in the mixture. We found that at various asparagines and citrulline ratios [= 3:1, 1:1, and 1:3; corresponding to 75:25, 50:50, and 25:75 (μMol ml(−1)/μMol ml(−1))], the resulting peak exhibited different retention times. Adjustment of ureide ratios as internal standards enables peak identification and quantification. Both chemicals were quantified in xylem sap samples of pecan [Carya illinoinensis (Wangenh.) K. Koch] trees. Analysis revealed that tree nickel nutrition status affects relative concentrations of Urea Cycle intermediates, asparagine and citrulline, present in sap. Consequently, we concluded that the HPLC methods are presented to enable qualitative and quantitative analysis of these metabolically important ureides.
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spelling pubmed-27168122009-08-06 Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC) Bai, Cheng Reilly, Charles C. Wood, Bruce W. Anal Chem Insights Original Research High-performance liquid chromatography (HPLC) analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates). Asparagine alone is eluted earlier than citrulline alone, but when both of them are present in biological samples they may co-elute. HPLC retention times for asparagine and citrulline were influenced by other ureides in the mixture. We found that at various asparagines and citrulline ratios [= 3:1, 1:1, and 1:3; corresponding to 75:25, 50:50, and 25:75 (μMol ml(−1)/μMol ml(−1))], the resulting peak exhibited different retention times. Adjustment of ureide ratios as internal standards enables peak identification and quantification. Both chemicals were quantified in xylem sap samples of pecan [Carya illinoinensis (Wangenh.) K. Koch] trees. Analysis revealed that tree nickel nutrition status affects relative concentrations of Urea Cycle intermediates, asparagine and citrulline, present in sap. Consequently, we concluded that the HPLC methods are presented to enable qualitative and quantitative analysis of these metabolically important ureides. Libertas Academica 2007-03-28 /pmc/articles/PMC2716812/ /pubmed/19662174 Text en Copyright © 2007 The authors. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Original Research
Bai, Cheng
Reilly, Charles C.
Wood, Bruce W.
Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)
title Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)
title_full Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)
title_fullStr Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)
title_full_unstemmed Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)
title_short Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)
title_sort identification and quantitation of asparagine and citrulline using high-performance liquid chromatography (hplc)
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2716812/
https://www.ncbi.nlm.nih.gov/pubmed/19662174
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