Increasing lipidomic coverage by selecting optimal mobile-phase modifiers in LC–MS of blood plasma

Tomas Cajka, Oliver Fiehn

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Reversed-phase ultrahigh-performance liquid chromatography–mass spectrometry (UHPLC–MS) is the method of choice for lipidomic profiling of blood plasma. For comprehensive screening, lipids need to be screened in both positive and negative electrospray mode. We here show that optimal results for increased lipidome coverage are only obtained if two different mobile-phase modifier systems are used, as opposed to using a single common modifier for both ionization modes. Specifically, ammonium acetate and ammonium formate were tested with and without acidifiers on standards representing 16 lipid classes, as well as on 164 blood plasma lipids from 11 lipid classes. Optimal coverage was obtained using 10 mM ammonium formate in ESI(+) and 10 mM ammonium acetate in ESI(−), both without acidification. Importantly, detection of free fatty acids was suppressed at more than 50-fold lower signal intensity if ammonium formate or formic acid as acidifier was used in ESI(−), whereas cholesteryl esters were hampered by up to 10-fold lower peak heights when ammonium acetate was used under ESI(+) conditions. Using charged surface hybrid C18 columns had slight advantages for lipids detected in blood plasma extracts, except for phosphatidic acids for which bridged ethylene hybrid C18 columns showed clear advantages.

Original languageEnglish (US)
Article number34
Pages (from-to)1-11
Number of pages11
JournalMetabolomics
Volume12
Issue number2
DOIs
StatePublished - Feb 1 2016

Fingerprint

formic acid
Blood
Lipids
Plasmas
Phosphatidic Acids
Acidification
Cholesterol Esters
Nonesterified Fatty Acids
Spectrometry
Ionization
Spectrum Analysis
Screening
Liquids

Keywords

  • Blood plasma
  • Lipidomics
  • Liquid chromatography–mass spectrometry
  • Mobile phase
  • Modifiers
  • Optimization

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Endocrinology, Diabetes and Metabolism

Cite this

Increasing lipidomic coverage by selecting optimal mobile-phase modifiers in LC–MS of blood plasma. / Cajka, Tomas; Fiehn, Oliver.

In: Metabolomics, Vol. 12, No. 2, 34, 01.02.2016, p. 1-11.

Research output: Contribution to journalArticle

@article{3f9a29f6e12c409ab383510ad551ad1d,
title = "Increasing lipidomic coverage by selecting optimal mobile-phase modifiers in LC–MS of blood plasma",
abstract = "Reversed-phase ultrahigh-performance liquid chromatography–mass spectrometry (UHPLC–MS) is the method of choice for lipidomic profiling of blood plasma. For comprehensive screening, lipids need to be screened in both positive and negative electrospray mode. We here show that optimal results for increased lipidome coverage are only obtained if two different mobile-phase modifier systems are used, as opposed to using a single common modifier for both ionization modes. Specifically, ammonium acetate and ammonium formate were tested with and without acidifiers on standards representing 16 lipid classes, as well as on 164 blood plasma lipids from 11 lipid classes. Optimal coverage was obtained using 10 mM ammonium formate in ESI(+) and 10 mM ammonium acetate in ESI(−), both without acidification. Importantly, detection of free fatty acids was suppressed at more than 50-fold lower signal intensity if ammonium formate or formic acid as acidifier was used in ESI(−), whereas cholesteryl esters were hampered by up to 10-fold lower peak heights when ammonium acetate was used under ESI(+) conditions. Using charged surface hybrid C18 columns had slight advantages for lipids detected in blood plasma extracts, except for phosphatidic acids for which bridged ethylene hybrid C18 columns showed clear advantages.",
keywords = "Blood plasma, Lipidomics, Liquid chromatography–mass spectrometry, Mobile phase, Modifiers, Optimization",
author = "Tomas Cajka and Oliver Fiehn",
year = "2016",
month = "2",
day = "1",
doi = "10.1007/s11306-015-0929-x",
language = "English (US)",
volume = "12",
pages = "1--11",
journal = "Metabolomics",
issn = "1573-3882",
publisher = "Springer New York",
number = "2",

}

TY - JOUR

T1 - Increasing lipidomic coverage by selecting optimal mobile-phase modifiers in LC–MS of blood plasma

AU - Cajka, Tomas

AU - Fiehn, Oliver

PY - 2016/2/1

Y1 - 2016/2/1

N2 - Reversed-phase ultrahigh-performance liquid chromatography–mass spectrometry (UHPLC–MS) is the method of choice for lipidomic profiling of blood plasma. For comprehensive screening, lipids need to be screened in both positive and negative electrospray mode. We here show that optimal results for increased lipidome coverage are only obtained if two different mobile-phase modifier systems are used, as opposed to using a single common modifier for both ionization modes. Specifically, ammonium acetate and ammonium formate were tested with and without acidifiers on standards representing 16 lipid classes, as well as on 164 blood plasma lipids from 11 lipid classes. Optimal coverage was obtained using 10 mM ammonium formate in ESI(+) and 10 mM ammonium acetate in ESI(−), both without acidification. Importantly, detection of free fatty acids was suppressed at more than 50-fold lower signal intensity if ammonium formate or formic acid as acidifier was used in ESI(−), whereas cholesteryl esters were hampered by up to 10-fold lower peak heights when ammonium acetate was used under ESI(+) conditions. Using charged surface hybrid C18 columns had slight advantages for lipids detected in blood plasma extracts, except for phosphatidic acids for which bridged ethylene hybrid C18 columns showed clear advantages.

AB - Reversed-phase ultrahigh-performance liquid chromatography–mass spectrometry (UHPLC–MS) is the method of choice for lipidomic profiling of blood plasma. For comprehensive screening, lipids need to be screened in both positive and negative electrospray mode. We here show that optimal results for increased lipidome coverage are only obtained if two different mobile-phase modifier systems are used, as opposed to using a single common modifier for both ionization modes. Specifically, ammonium acetate and ammonium formate were tested with and without acidifiers on standards representing 16 lipid classes, as well as on 164 blood plasma lipids from 11 lipid classes. Optimal coverage was obtained using 10 mM ammonium formate in ESI(+) and 10 mM ammonium acetate in ESI(−), both without acidification. Importantly, detection of free fatty acids was suppressed at more than 50-fold lower signal intensity if ammonium formate or formic acid as acidifier was used in ESI(−), whereas cholesteryl esters were hampered by up to 10-fold lower peak heights when ammonium acetate was used under ESI(+) conditions. Using charged surface hybrid C18 columns had slight advantages for lipids detected in blood plasma extracts, except for phosphatidic acids for which bridged ethylene hybrid C18 columns showed clear advantages.

KW - Blood plasma

KW - Lipidomics

KW - Liquid chromatography–mass spectrometry

KW - Mobile phase

KW - Modifiers

KW - Optimization

UR - http://www.scopus.com/inward/record.url?scp=84954459129&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84954459129&partnerID=8YFLogxK

U2 - 10.1007/s11306-015-0929-x

DO - 10.1007/s11306-015-0929-x

M3 - Article

VL - 12

SP - 1

EP - 11

JO - Metabolomics

JF - Metabolomics

SN - 1573-3882

IS - 2

M1 - 34

ER -