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Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood

Given that the peculiar redox behavior of ergothioneine involves a rapid regeneration process, the measurement of its precursor and redox metabolite hercynine could be particularly useful in assessing its role in oxidative stress or other biological processes. Thus, a LC-MS/MS method for the determi...

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Autores principales: Sotgia, Salvatore, Murphy, Rhys B., Zinellu, Angelo, Elliot, David, Paliogiannis, Panagiotis, Pinna, Gerard Aimè, Carru, Ciriaco, Mangoni, Arduino A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6321398/
https://www.ncbi.nlm.nih.gov/pubmed/30558219
http://dx.doi.org/10.3390/molecules23123326
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author Sotgia, Salvatore
Murphy, Rhys B.
Zinellu, Angelo
Elliot, David
Paliogiannis, Panagiotis
Pinna, Gerard Aimè
Carru, Ciriaco
Mangoni, Arduino A.
author_facet Sotgia, Salvatore
Murphy, Rhys B.
Zinellu, Angelo
Elliot, David
Paliogiannis, Panagiotis
Pinna, Gerard Aimè
Carru, Ciriaco
Mangoni, Arduino A.
author_sort Sotgia, Salvatore
collection PubMed
description Given that the peculiar redox behavior of ergothioneine involves a rapid regeneration process, the measurement of its precursor and redox metabolite hercynine could be particularly useful in assessing its role in oxidative stress or other biological processes. Thus, a LC-MS/MS method for the determination of hercynine concentrations in whole blood was developed. After lysis of red blood cells by cold water, samples were filtered on micro concentrators at a controlled temperature of 4 °C. The clear filtered fluid was then treated with diethylpyrocarbonate to derivatize hercynine for the analysis by LC-MS/MS. The derivatized analyte was isocratically separated as a carbethoxy derivative on a C18 column with a mobile phase of an aqueous 0.1% v/v formic acid and acetonitrile (95:5). Effluents were monitored by MRM transitions at m/z 270.28→95 and 273.21→95 for hercynine and its deuterated counterpart, respectively. No cross-talk between MRM transitions was observed and a good linearity was found within a range of 35–1120 nmol/L. The LOD and LOQ were, respectively, 10.30 and 31.21 nmol/L with an intraday and intermediate precision below 7%. The average hercynine concentration in whole blood from 30 healthy male volunteers (aged 77 ± 12 years) was 178.5 ± 118.1 nmol/L. Overall, the method is easy to perform, allowing a rapid and accurate assessment of whole blood concentrations of hercynine.
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spelling pubmed-63213982019-01-14 Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood Sotgia, Salvatore Murphy, Rhys B. Zinellu, Angelo Elliot, David Paliogiannis, Panagiotis Pinna, Gerard Aimè Carru, Ciriaco Mangoni, Arduino A. Molecules Article Given that the peculiar redox behavior of ergothioneine involves a rapid regeneration process, the measurement of its precursor and redox metabolite hercynine could be particularly useful in assessing its role in oxidative stress or other biological processes. Thus, a LC-MS/MS method for the determination of hercynine concentrations in whole blood was developed. After lysis of red blood cells by cold water, samples were filtered on micro concentrators at a controlled temperature of 4 °C. The clear filtered fluid was then treated with diethylpyrocarbonate to derivatize hercynine for the analysis by LC-MS/MS. The derivatized analyte was isocratically separated as a carbethoxy derivative on a C18 column with a mobile phase of an aqueous 0.1% v/v formic acid and acetonitrile (95:5). Effluents were monitored by MRM transitions at m/z 270.28→95 and 273.21→95 for hercynine and its deuterated counterpart, respectively. No cross-talk between MRM transitions was observed and a good linearity was found within a range of 35–1120 nmol/L. The LOD and LOQ were, respectively, 10.30 and 31.21 nmol/L with an intraday and intermediate precision below 7%. The average hercynine concentration in whole blood from 30 healthy male volunteers (aged 77 ± 12 years) was 178.5 ± 118.1 nmol/L. Overall, the method is easy to perform, allowing a rapid and accurate assessment of whole blood concentrations of hercynine. MDPI 2018-12-14 /pmc/articles/PMC6321398/ /pubmed/30558219 http://dx.doi.org/10.3390/molecules23123326 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sotgia, Salvatore
Murphy, Rhys B.
Zinellu, Angelo
Elliot, David
Paliogiannis, Panagiotis
Pinna, Gerard Aimè
Carru, Ciriaco
Mangoni, Arduino A.
Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood
title Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood
title_full Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood
title_fullStr Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood
title_full_unstemmed Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood
title_short Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood
title_sort development of an lc–tandem mass spectrometry method for the quantitative analysis of hercynine in human whole blood
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6321398/
https://www.ncbi.nlm.nih.gov/pubmed/30558219
http://dx.doi.org/10.3390/molecules23123326
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