Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS

<b> </b>Kidneys play a crucial role in maintaining metabolic homeostasis in a body. Serum creatinine concentration is a simple test used as an indicator of renal function. One of the known ways of quantifying creatinine concentration is the liquid chromatography-mass spectrometry (LC-MS)...

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Main Authors: Remigiusz Bąchor, Andrzej Konieczny, Zbigniew Szewczuk
Format: Article
Language:English
Published: MDPI AG 2020-03-01
Series:Molecules
Subjects:
Online Access:https://www.mdpi.com/1420-3049/25/7/1514
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author Remigiusz Bąchor
Andrzej Konieczny
Zbigniew Szewczuk
author_facet Remigiusz Bąchor
Andrzej Konieczny
Zbigniew Szewczuk
author_sort Remigiusz Bąchor
collection DOAJ
description <b> </b>Kidneys play a crucial role in maintaining metabolic homeostasis in a body. Serum creatinine concentration is a simple test used as an indicator of renal function. One of the known ways of quantifying creatinine concentration is the liquid chromatography-mass spectrometry (LC-MS) method, using an isotopically labeled analog of creatinine as an internal standard. Unfortunately, such isotope-labeled analogs are expensive and their synthesis is complex. Here we demonstrate a facile preparation of deuterated analogues of creatinine, via the H/D exchange of hydrogens located at the &#945;-carbon (&#945;-C) of the N-methylated amino acid part, under basic conditions. The stability of retrieved isotopologues was analyzed under both neutral or acidic conditions, and the results revealed that the introduced deuterons do not undergo back-exchange. In addition, the coelution of deuterated and non-deuterated forms under acidic and neutral conditions was observed. The prepared isotopologues were successfully applied in the quantitative LC-MS analysis of urine samples, and the results demonstrated that the presented strategy is novel and inexpensive, and that the quantification correlates with the commonly used Jaffe test method.
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spelling doaj.art-278c280c03964101a5db8dd00a70da022022-12-21T20:26:26ZengMDPI AGMolecules1420-30492020-03-01257151410.3390/molecules25071514molecules25071514Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MSRemigiusz Bąchor0Andrzej Konieczny1Zbigniew Szewczuk2Faculty of Chemistry, University of Wroclaw, 50-383 Wroclaw, PolandDepartment of Nephrology and Transplantation Medicine, Wroclaw Medical University, 50-556 Wroclaw, PolandFaculty of Chemistry, University of Wroclaw, 50-383 Wroclaw, Poland<b> </b>Kidneys play a crucial role in maintaining metabolic homeostasis in a body. Serum creatinine concentration is a simple test used as an indicator of renal function. One of the known ways of quantifying creatinine concentration is the liquid chromatography-mass spectrometry (LC-MS) method, using an isotopically labeled analog of creatinine as an internal standard. Unfortunately, such isotope-labeled analogs are expensive and their synthesis is complex. Here we demonstrate a facile preparation of deuterated analogues of creatinine, via the H/D exchange of hydrogens located at the &#945;-carbon (&#945;-C) of the N-methylated amino acid part, under basic conditions. The stability of retrieved isotopologues was analyzed under both neutral or acidic conditions, and the results revealed that the introduced deuterons do not undergo back-exchange. In addition, the coelution of deuterated and non-deuterated forms under acidic and neutral conditions was observed. The prepared isotopologues were successfully applied in the quantitative LC-MS analysis of urine samples, and the results demonstrated that the presented strategy is novel and inexpensive, and that the quantification correlates with the commonly used Jaffe test method.https://www.mdpi.com/1420-3049/25/7/1514creatininehydrogen/deuterium exchangeisotope dilutionliquid chromatography-mass spectrometry
spellingShingle Remigiusz Bąchor
Andrzej Konieczny
Zbigniew Szewczuk
Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS
Molecules
creatinine
hydrogen/deuterium exchange
isotope dilution
liquid chromatography-mass spectrometry
title Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS
title_full Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS
title_fullStr Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS
title_full_unstemmed Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS
title_short Preparation of Isotopically Labelled Standards of Creatinine Via H/D Exchange and Their Application in Quantitative Analysis by LC-MS
title_sort preparation of isotopically labelled standards of creatinine via h d exchange and their application in quantitative analysis by lc ms
topic creatinine
hydrogen/deuterium exchange
isotope dilution
liquid chromatography-mass spectrometry
url https://www.mdpi.com/1420-3049/25/7/1514
work_keys_str_mv AT remigiuszbachor preparationofisotopicallylabelledstandardsofcreatinineviahdexchangeandtheirapplicationinquantitativeanalysisbylcms
AT andrzejkonieczny preparationofisotopicallylabelledstandardsofcreatinineviahdexchangeandtheirapplicationinquantitativeanalysisbylcms
AT zbigniewszewczuk preparationofisotopicallylabelledstandardsofcreatinineviahdexchangeandtheirapplicationinquantitativeanalysisbylcms