An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues

Abstract Ammonia is a fundamental aspect of metabolism spanning all of phylogeny. Metabolomics, including metabolic tracing studies, are an integral part of elucidating the role of ammonia in these systems. However, current methods for measurement of ammonia are spectrophotometric, and cannot distin...

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Autores principales:	Jessica B. Spinelli, Liam P. Kelley, Marcia C. Haigis
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7eaa85d8b8bd4cc5b2626a94276327df
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spelling	oai:doaj.org-article:7eaa85d8b8bd4cc5b2626a94276327df2021-12-02T15:06:13ZAn LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues10.1038/s41598-017-09993-62045-2322https://doaj.org/article/7eaa85d8b8bd4cc5b2626a94276327df2017-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-09993-6https://doaj.org/toc/2045-2322Abstract Ammonia is a fundamental aspect of metabolism spanning all of phylogeny. Metabolomics, including metabolic tracing studies, are an integral part of elucidating the role of ammonia in these systems. However, current methods for measurement of ammonia are spectrophotometric, and cannot distinguish isotopologues of ammonia, significantly limiting metabolic tracing studies. Here, we describe a novel LC-MS-based method that quantitatively assesses both 14N-and 15N-isotopologues of ammonia in polar metabolite extracts. This assay (1) quantitatively measures the concentration of ammonia in polar metabolite isolates used for metabolomic studies, and (2) accurately determines the percent isotope abundance of 15N-ammonia in a cell lysate for 15N-isotope tracing studies. We apply this assay to quantitatively measure glutamine-derived ammonia in lung cancer cell lines with differential expression of glutaminase.Jessica B. SpinelliLiam P. KelleyMarcia C. HaigisNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-8 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jessica B. Spinelli Liam P. Kelley Marcia C. Haigis An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues
description	Abstract Ammonia is a fundamental aspect of metabolism spanning all of phylogeny. Metabolomics, including metabolic tracing studies, are an integral part of elucidating the role of ammonia in these systems. However, current methods for measurement of ammonia are spectrophotometric, and cannot distinguish isotopologues of ammonia, significantly limiting metabolic tracing studies. Here, we describe a novel LC-MS-based method that quantitatively assesses both 14N-and 15N-isotopologues of ammonia in polar metabolite extracts. This assay (1) quantitatively measures the concentration of ammonia in polar metabolite isolates used for metabolomic studies, and (2) accurately determines the percent isotope abundance of 15N-ammonia in a cell lysate for 15N-isotope tracing studies. We apply this assay to quantitatively measure glutamine-derived ammonia in lung cancer cell lines with differential expression of glutaminase.
format	article
author	Jessica B. Spinelli Liam P. Kelley Marcia C. Haigis
author_facet	Jessica B. Spinelli Liam P. Kelley Marcia C. Haigis
author_sort	Jessica B. Spinelli
title	An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues
title_short	An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues
title_full	An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues
title_fullStr	An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues
title_full_unstemmed	An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues
title_sort	lc-ms approach to quantitative measurement of ammonia isotopologues
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/7eaa85d8b8bd4cc5b2626a94276327df
work_keys_str_mv	AT jessicabspinelli anlcmsapproachtoquantitativemeasurementofammoniaisotopologues AT liampkelley anlcmsapproachtoquantitativemeasurementofammoniaisotopologues AT marciachaigis anlcmsapproachtoquantitativemeasurementofammoniaisotopologues AT jessicabspinelli lcmsapproachtoquantitativemeasurementofammoniaisotopologues AT liampkelley lcmsapproachtoquantitativemeasurementofammoniaisotopologues AT marciachaigis lcmsapproachtoquantitativemeasurementofammoniaisotopologues
_version_	1718388523923406848

An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues

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