Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma
Polar hydrophilic metabolites have been identified as important actors in many biochemical pathways. Despite continuous improvement and refinement of hydrophilic interaction liquid chromatography (HILIC) platforms, its application in global polar metabolomics has been underutilized. In this study, w...
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MDPI AG
2022-02-01
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Online Access: | https://www.mdpi.com/2218-1989/12/2/165 |
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author | Farideh Hosseinkhani Luojiao Huang Anne-Charlotte Dubbelman Faisa Guled Amy C. Harms Thomas Hankemeier |
author_facet | Farideh Hosseinkhani Luojiao Huang Anne-Charlotte Dubbelman Faisa Guled Amy C. Harms Thomas Hankemeier |
author_sort | Farideh Hosseinkhani |
collection | DOAJ |
description | Polar hydrophilic metabolites have been identified as important actors in many biochemical pathways. Despite continuous improvement and refinement of hydrophilic interaction liquid chromatography (HILIC) platforms, its application in global polar metabolomics has been underutilized. In this study, we aimed to systematically evaluate polar stationary phases for untargeted metabolomics by using HILIC columns (neutral and zwitterionic) that have been exploited widely in targeted approaches. To do so, high-resolution mass spectrometry was applied to thoroughly investigate selectivity, repeatability and matrix effect at three pH conditions for 9 classes of polar compounds using 54 authentic standards and plasma matrix. The column performance for utilization in untargeted metabolomics was assessed using plasma samples with diverse phenotypes. Our results indicate that the ZIC-c HILIC column operated at neutral pH exhibited several advantages, including superior performance for different classes of compounds, better isomer separation, repeatability and high metabolic coverage. Regardless of the column type, the retention of inorganic ions in plasma leads to extensive adduct formation and co-elution with analytes, which results in ion-suppression as part of the overall plasma matrix effect. In ZIC-c HILIC, the sodium chloride ion effect was particularly observed for amino acids and amine classes. Successful performance of HILIC for separation of plasma samples with different phenotypes highlights this mode of separation as a valuable approach in global profiling of plasma sample and discovering the metabolic changes associated with health and disease. |
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issn | 2218-1989 |
language | English |
last_indexed | 2024-03-09T21:27:36Z |
publishDate | 2022-02-01 |
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spelling | doaj.art-7af911bd6f4e4dc09228a16a579ccf9a2023-11-23T21:05:25ZengMDPI AGMetabolites2218-19892022-02-0112216510.3390/metabo12020165Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human PlasmaFarideh Hosseinkhani0Luojiao Huang1Anne-Charlotte Dubbelman2Faisa Guled3Amy C. Harms4Thomas Hankemeier5Leiden Academic Centre for Drug Research, Division of Systems Biomedicine and Pharmacology, Leiden University, 2300 RA Leiden, The NetherlandsLeiden Academic Centre for Drug Research, Division of Systems Biomedicine and Pharmacology, Leiden University, 2300 RA Leiden, The NetherlandsLeiden Academic Centre for Drug Research, Division of Systems Biomedicine and Pharmacology, Leiden University, 2300 RA Leiden, The NetherlandsLeiden Academic Centre for Drug Research, Division of Systems Biomedicine and Pharmacology, Leiden University, 2300 RA Leiden, The NetherlandsLeiden Academic Centre for Drug Research, Division of Systems Biomedicine and Pharmacology, Leiden University, 2300 RA Leiden, The NetherlandsLeiden Academic Centre for Drug Research, Division of Systems Biomedicine and Pharmacology, Leiden University, 2300 RA Leiden, The NetherlandsPolar hydrophilic metabolites have been identified as important actors in many biochemical pathways. Despite continuous improvement and refinement of hydrophilic interaction liquid chromatography (HILIC) platforms, its application in global polar metabolomics has been underutilized. In this study, we aimed to systematically evaluate polar stationary phases for untargeted metabolomics by using HILIC columns (neutral and zwitterionic) that have been exploited widely in targeted approaches. To do so, high-resolution mass spectrometry was applied to thoroughly investigate selectivity, repeatability and matrix effect at three pH conditions for 9 classes of polar compounds using 54 authentic standards and plasma matrix. The column performance for utilization in untargeted metabolomics was assessed using plasma samples with diverse phenotypes. Our results indicate that the ZIC-c HILIC column operated at neutral pH exhibited several advantages, including superior performance for different classes of compounds, better isomer separation, repeatability and high metabolic coverage. Regardless of the column type, the retention of inorganic ions in plasma leads to extensive adduct formation and co-elution with analytes, which results in ion-suppression as part of the overall plasma matrix effect. In ZIC-c HILIC, the sodium chloride ion effect was particularly observed for amino acids and amine classes. Successful performance of HILIC for separation of plasma samples with different phenotypes highlights this mode of separation as a valuable approach in global profiling of plasma sample and discovering the metabolic changes associated with health and disease.https://www.mdpi.com/2218-1989/12/2/165HILICuntargeted metabolomicspolar metabolitesplasma metabolomicsliquid chromatography |
spellingShingle | Farideh Hosseinkhani Luojiao Huang Anne-Charlotte Dubbelman Faisa Guled Amy C. Harms Thomas Hankemeier Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma Metabolites HILIC untargeted metabolomics polar metabolites plasma metabolomics liquid chromatography |
title | Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma |
title_full | Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma |
title_fullStr | Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma |
title_full_unstemmed | Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma |
title_short | Systematic Evaluation of HILIC Stationary Phases for Global Metabolomics of Human Plasma |
title_sort | systematic evaluation of hilic stationary phases for global metabolomics of human plasma |
topic | HILIC untargeted metabolomics polar metabolites plasma metabolomics liquid chromatography |
url | https://www.mdpi.com/2218-1989/12/2/165 |
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