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In-vial dual extraction liquid chromatography coupled to mass spectrometry applied to streptozotocin-treated diabetic rats. Tips and pitfalls of the method

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@article{37641857f8984904b91c00adb09b7b20,
title = "In-vial dual extraction liquid chromatography coupled to mass spectrometry applied to streptozotocin-treated diabetic rats. Tips and pitfalls of the method",
abstract = "The aim of metabolomics studies is the comprehensive and quantitative analysis of all metabolites in a cell, tissue or organism. This approach requires sample preparation methods to be fast, reproducible and able to extract a wide range of analytes with different polarities, as well as analytical platforms able to detect the extracted metabolites. Recently, we have developed a one-step extraction method consisting of a lipophilic and hydrophilic layer within a single vial insert, in-vial dual extraction (IVDE). In order to check possible application of this method to real biological case, analysis of plasma samples obtained from three streptozotocin-induced diabetic and three control rats was performed. Analytical validity of the method was proved by the calculation (in quality control samples) of relative standard deviation (RSD) for detected metabolites. The percentage of metabolites with RSD<30{\%} was 93{\%} for Fatty acyls, 80{\%} for Glycerolipids, 93{\%} for Glycerophospholipids, 68{\%} for Sterol lipids, and 91{\%} for Sphingolipids. IVDE allowed for selection of more than 600 different features discriminating two studied groups. For around 40{\%} of these masses putative identification was possible. Adequate, with several considerations described within this paper, application of IVDE method enables wide metabolite coverage from a single 20μL plasma aliquot. Within the features putatively identified, glycerolipids and glycerophospholipids arose as the most important groups of compounds discriminating diabetic rats from controls. All discriminating metabolites give an idea of the large metabolic differences that can be present in non-controlled type 1 diabetes.",
keywords = "Animals, Chromatography, High Pressure Liquid/methods, Diabetes Mellitus, Experimental/blood, Lipid Metabolism, Lipids/blood, Male, Mass Spectrometry/methods, Metabolomics/methods, Rats, Rats, Sprague-Dawley",
author = "Joanna Godzien and Michal Ciborowski and Luke Whiley and Ruperez, {Francisco J} and Coral Barbas and Cristina Legido-Quigley",
note = "Copyright {\circledC} 2013 Elsevier B.V. All rights reserved.",
year = "2013",
month = "8",
day = "23",
doi = "10.1016/j.chroma.2013.07.029",
language = "English",
volume = "1304",
pages = "52--60",
journal = "Journal of Chromatography",
issn = "0021-9673",
publisher = "Elsevier BV",

}

RIS

TY - JOUR

T1 - In-vial dual extraction liquid chromatography coupled to mass spectrometry applied to streptozotocin-treated diabetic rats. Tips and pitfalls of the method

AU - Godzien, Joanna

AU - Ciborowski, Michal

AU - Whiley, Luke

AU - Ruperez, Francisco J

AU - Barbas, Coral

AU - Legido-Quigley, Cristina

N1 - Copyright © 2013 Elsevier B.V. All rights reserved.

PY - 2013/8/23

Y1 - 2013/8/23

N2 - The aim of metabolomics studies is the comprehensive and quantitative analysis of all metabolites in a cell, tissue or organism. This approach requires sample preparation methods to be fast, reproducible and able to extract a wide range of analytes with different polarities, as well as analytical platforms able to detect the extracted metabolites. Recently, we have developed a one-step extraction method consisting of a lipophilic and hydrophilic layer within a single vial insert, in-vial dual extraction (IVDE). In order to check possible application of this method to real biological case, analysis of plasma samples obtained from three streptozotocin-induced diabetic and three control rats was performed. Analytical validity of the method was proved by the calculation (in quality control samples) of relative standard deviation (RSD) for detected metabolites. The percentage of metabolites with RSD<30% was 93% for Fatty acyls, 80% for Glycerolipids, 93% for Glycerophospholipids, 68% for Sterol lipids, and 91% for Sphingolipids. IVDE allowed for selection of more than 600 different features discriminating two studied groups. For around 40% of these masses putative identification was possible. Adequate, with several considerations described within this paper, application of IVDE method enables wide metabolite coverage from a single 20μL plasma aliquot. Within the features putatively identified, glycerolipids and glycerophospholipids arose as the most important groups of compounds discriminating diabetic rats from controls. All discriminating metabolites give an idea of the large metabolic differences that can be present in non-controlled type 1 diabetes.

AB - The aim of metabolomics studies is the comprehensive and quantitative analysis of all metabolites in a cell, tissue or organism. This approach requires sample preparation methods to be fast, reproducible and able to extract a wide range of analytes with different polarities, as well as analytical platforms able to detect the extracted metabolites. Recently, we have developed a one-step extraction method consisting of a lipophilic and hydrophilic layer within a single vial insert, in-vial dual extraction (IVDE). In order to check possible application of this method to real biological case, analysis of plasma samples obtained from three streptozotocin-induced diabetic and three control rats was performed. Analytical validity of the method was proved by the calculation (in quality control samples) of relative standard deviation (RSD) for detected metabolites. The percentage of metabolites with RSD<30% was 93% for Fatty acyls, 80% for Glycerolipids, 93% for Glycerophospholipids, 68% for Sterol lipids, and 91% for Sphingolipids. IVDE allowed for selection of more than 600 different features discriminating two studied groups. For around 40% of these masses putative identification was possible. Adequate, with several considerations described within this paper, application of IVDE method enables wide metabolite coverage from a single 20μL plasma aliquot. Within the features putatively identified, glycerolipids and glycerophospholipids arose as the most important groups of compounds discriminating diabetic rats from controls. All discriminating metabolites give an idea of the large metabolic differences that can be present in non-controlled type 1 diabetes.

KW - Animals

KW - Chromatography, High Pressure Liquid/methods

KW - Diabetes Mellitus, Experimental/blood

KW - Lipid Metabolism

KW - Lipids/blood

KW - Male

KW - Mass Spectrometry/methods

KW - Metabolomics/methods

KW - Rats

KW - Rats, Sprague-Dawley

U2 - 10.1016/j.chroma.2013.07.029

DO - 10.1016/j.chroma.2013.07.029

M3 - Journal article

VL - 1304

SP - 52

EP - 60

JO - Journal of Chromatography

JF - Journal of Chromatography

SN - 0021-9673

ER -

ID: 54960961