Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis

In recent years, there has been a growing demand for making the analytical processes sustainable and environmentally friendly. In proteomic analysis, sample preparation plays an important role, employing different extraction techniques to isolate proteins of interest from biological fluids to be abl...

Full description

Saved in:
Bibliographic Details
Main Authors: Martina Opetová, Radovan Tomašovský, Katarína Maráková
Format: Article
Language:English
Published: Elsevier 2025-02-01
Series:Advances in Sample Preparation
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2772582025000130
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849723151614738432
author Martina Opetová
Radovan Tomašovský
Katarína Maráková
author_facet Martina Opetová
Radovan Tomašovský
Katarína Maráková
author_sort Martina Opetová
collection DOAJ
description In recent years, there has been a growing demand for making the analytical processes sustainable and environmentally friendly. In proteomic analysis, sample preparation plays an important role, employing different extraction techniques to isolate proteins of interest from biological fluids to be able to analyze them. These techniques usually use large volumes of organic solvents, such as acetonitrile, which has been recognized as hazardous and difficult to dispense. According to green analytical chemistry principles, the environmental impact of the methods can be reduced either by miniaturizing the processes or by replacing organic solvents with less toxic alternatives. This work compares the effect of different greener solvents on the solid-phase extraction process of low molecular weight proteins spiked in biological fluids (pooled human serum, plasma, urine, and saliva) and suggests that ethanol (60 %, v/v) offers, when used as an elution solvent, comparable results to acetonitrile (75 % v/v) in terms of extraction recovery. The addition of acidic modifiers to the elution solvent did not improve the extraction recovery, and the nitrogen evaporation of the eluate resulted in a 17–53 % loss of the proteins compared to the simple dilution of the eluate.
format Article
id doaj-art-a4abb91b8ab74d5b9fc533b7a562fb17
institution DOAJ
issn 2772-5820
language English
publishDate 2025-02-01
publisher Elsevier
record_format Article
series Advances in Sample Preparation
spelling doaj-art-a4abb91b8ab74d5b9fc533b7a562fb172025-08-20T03:11:06ZengElsevierAdvances in Sample Preparation2772-58202025-02-011310016010.1016/j.sampre.2025.100160Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysisMartina Opetová0Radovan Tomašovský1Katarína Maráková2Comenius University Bratislava, Faculty of Pharmacy, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Odbojárov 10, SK-832 32, Bratislava, Slovakia; Comenius University Bratislava, Faculty of Pharmacy, Toxicological and Antidoping Center, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Odbojárov 10, SK-832 32, Bratislava, SlovakiaComenius University Bratislava, Faculty of Pharmacy, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Odbojárov 10, SK-832 32, Bratislava, Slovakia; Comenius University Bratislava, Faculty of Pharmacy, Toxicological and Antidoping Center, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Odbojárov 10, SK-832 32, Bratislava, SlovakiaComenius University Bratislava, Faculty of Pharmacy, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Odbojárov 10, SK-832 32, Bratislava, Slovakia; Comenius University Bratislava, Faculty of Pharmacy, Toxicological and Antidoping Center, Department of Pharmaceutical Analysis and Nuclear Pharmacy, Odbojárov 10, SK-832 32, Bratislava, Slovakia; Corresponding author.In recent years, there has been a growing demand for making the analytical processes sustainable and environmentally friendly. In proteomic analysis, sample preparation plays an important role, employing different extraction techniques to isolate proteins of interest from biological fluids to be able to analyze them. These techniques usually use large volumes of organic solvents, such as acetonitrile, which has been recognized as hazardous and difficult to dispense. According to green analytical chemistry principles, the environmental impact of the methods can be reduced either by miniaturizing the processes or by replacing organic solvents with less toxic alternatives. This work compares the effect of different greener solvents on the solid-phase extraction process of low molecular weight proteins spiked in biological fluids (pooled human serum, plasma, urine, and saliva) and suggests that ethanol (60 %, v/v) offers, when used as an elution solvent, comparable results to acetonitrile (75 % v/v) in terms of extraction recovery. The addition of acidic modifiers to the elution solvent did not improve the extraction recovery, and the nitrogen evaporation of the eluate resulted in a 17–53 % loss of the proteins compared to the simple dilution of the eluate.http://www.sciencedirect.com/science/article/pii/S2772582025000130Sample preparationCapillary electrophoresisIntact proteinsGreen solventsEthanol
spellingShingle Martina Opetová
Radovan Tomašovský
Katarína Maráková
Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis
Advances in Sample Preparation
Sample preparation
Capillary electrophoresis
Intact proteins
Green solvents
Ethanol
title Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis
title_full Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis
title_fullStr Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis
title_full_unstemmed Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis
title_short Greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top-down CE-MS analysis
title_sort greener solvents for microelution solid phase extraction of proteins from biological fluids followed by their top down ce ms analysis
topic Sample preparation
Capillary electrophoresis
Intact proteins
Green solvents
Ethanol
url http://www.sciencedirect.com/science/article/pii/S2772582025000130
work_keys_str_mv AT martinaopetova greenersolventsformicroelutionsolidphaseextractionofproteinsfrombiologicalfluidsfollowedbytheirtopdowncemsanalysis
AT radovantomasovsky greenersolventsformicroelutionsolidphaseextractionofproteinsfrombiologicalfluidsfollowedbytheirtopdowncemsanalysis
AT katarinamarakova greenersolventsformicroelutionsolidphaseextractionofproteinsfrombiologicalfluidsfollowedbytheirtopdowncemsanalysis