Cargando…

Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation

The aim of this study was to develop and optimize a chiral HPLC-MS/MS method for quantitative analysis of (R)-/(S)-salbutamol and (R)-/(S)-salbutamol-4′-O-sulfate in human urine to allow for bioanalytical quantitation of the targeted analytes and investigations of stereoselectivity in the sulfonatio...

Descripción completa

Detalles Bibliográficos
Autores principales: Harps, Lukas Corbinian, Jendretzki, Annika Lisa, Wolf, Clemens Alexander, Girreser, Ulrich, Wolber, Gerhard, Parr, Maria Kristina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10609612/
https://www.ncbi.nlm.nih.gov/pubmed/37894685
http://dx.doi.org/10.3390/molecules28207206
_version_ 1785128054374268928
author Harps, Lukas Corbinian
Jendretzki, Annika Lisa
Wolf, Clemens Alexander
Girreser, Ulrich
Wolber, Gerhard
Parr, Maria Kristina
author_facet Harps, Lukas Corbinian
Jendretzki, Annika Lisa
Wolf, Clemens Alexander
Girreser, Ulrich
Wolber, Gerhard
Parr, Maria Kristina
author_sort Harps, Lukas Corbinian
collection PubMed
description The aim of this study was to develop and optimize a chiral HPLC-MS/MS method for quantitative analysis of (R)-/(S)-salbutamol and (R)-/(S)-salbutamol-4′-O-sulfate in human urine to allow for bioanalytical quantitation of the targeted analytes and investigations of stereoselectivity in the sulfonation pathway of human phase Ⅱ metabolism. For analytical method development, a systematic screening of columns and mobile phases to develop a separation via enantiomerically selective high performance liquid chromatography was performed. Electrospray ionization settings were optimized via multiple-step screening and a full factorial design-of-experiment. Both approaches were performed matrix-assisted and the predicted values were compared. The full factorial design was superior in terms of prediction power and knowledge generation. Performing a longitudinal excretion study in one healthy volunteer allowed for the calculation of excretion rates for all four targeted analytes. For this proof-of-concept, either racemic salbutamol or enantiopure levosalbutamol was administered perorally or via inhalation, respectively. A strong preference for sulfonation of (R)-salbutamol for inhalation and peroral application was found in in vivo experiments. In previous studies phenol sulfotransferase 1A3 was described to be mainly responsible for salbutamol sulfonation in humans. Thus, in vitro and in silico investigations of the stereoselectivity of sulfotransferase 1A3 complemented the study and confirmed these findings.
format Online
Article
Text
id pubmed-10609612
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-106096122023-10-28 Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation Harps, Lukas Corbinian Jendretzki, Annika Lisa Wolf, Clemens Alexander Girreser, Ulrich Wolber, Gerhard Parr, Maria Kristina Molecules Article The aim of this study was to develop and optimize a chiral HPLC-MS/MS method for quantitative analysis of (R)-/(S)-salbutamol and (R)-/(S)-salbutamol-4′-O-sulfate in human urine to allow for bioanalytical quantitation of the targeted analytes and investigations of stereoselectivity in the sulfonation pathway of human phase Ⅱ metabolism. For analytical method development, a systematic screening of columns and mobile phases to develop a separation via enantiomerically selective high performance liquid chromatography was performed. Electrospray ionization settings were optimized via multiple-step screening and a full factorial design-of-experiment. Both approaches were performed matrix-assisted and the predicted values were compared. The full factorial design was superior in terms of prediction power and knowledge generation. Performing a longitudinal excretion study in one healthy volunteer allowed for the calculation of excretion rates for all four targeted analytes. For this proof-of-concept, either racemic salbutamol or enantiopure levosalbutamol was administered perorally or via inhalation, respectively. A strong preference for sulfonation of (R)-salbutamol for inhalation and peroral application was found in in vivo experiments. In previous studies phenol sulfotransferase 1A3 was described to be mainly responsible for salbutamol sulfonation in humans. Thus, in vitro and in silico investigations of the stereoselectivity of sulfotransferase 1A3 complemented the study and confirmed these findings. MDPI 2023-10-21 /pmc/articles/PMC10609612/ /pubmed/37894685 http://dx.doi.org/10.3390/molecules28207206 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Harps, Lukas Corbinian
Jendretzki, Annika Lisa
Wolf, Clemens Alexander
Girreser, Ulrich
Wolber, Gerhard
Parr, Maria Kristina
Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation
title Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation
title_full Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation
title_fullStr Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation
title_full_unstemmed Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation
title_short Development of an HPLC-MS/MS Method for Chiral Separation and Quantitation of (R)- and (S)-Salbutamol and Their Sulfoconjugated Metabolites in Urine to Investigate Stereoselective Sulfonation
title_sort development of an hplc-ms/ms method for chiral separation and quantitation of (r)- and (s)-salbutamol and their sulfoconjugated metabolites in urine to investigate stereoselective sulfonation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10609612/
https://www.ncbi.nlm.nih.gov/pubmed/37894685
http://dx.doi.org/10.3390/molecules28207206
work_keys_str_mv AT harpslukascorbinian developmentofanhplcmsmsmethodforchiralseparationandquantitationofrandssalbutamolandtheirsulfoconjugatedmetabolitesinurinetoinvestigatestereoselectivesulfonation
AT jendretzkiannikalisa developmentofanhplcmsmsmethodforchiralseparationandquantitationofrandssalbutamolandtheirsulfoconjugatedmetabolitesinurinetoinvestigatestereoselectivesulfonation
AT wolfclemensalexander developmentofanhplcmsmsmethodforchiralseparationandquantitationofrandssalbutamolandtheirsulfoconjugatedmetabolitesinurinetoinvestigatestereoselectivesulfonation
AT girreserulrich developmentofanhplcmsmsmethodforchiralseparationandquantitationofrandssalbutamolandtheirsulfoconjugatedmetabolitesinurinetoinvestigatestereoselectivesulfonation
AT wolbergerhard developmentofanhplcmsmsmethodforchiralseparationandquantitationofrandssalbutamolandtheirsulfoconjugatedmetabolitesinurinetoinvestigatestereoselectivesulfonation
AT parrmariakristina developmentofanhplcmsmsmethodforchiralseparationandquantitationofrandssalbutamolandtheirsulfoconjugatedmetabolitesinurinetoinvestigatestereoselectivesulfonation