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Development of an Automated Multi-Injection Shotgun Lipidomics Approach Using a Triple Quadrupole Mass Spectrometer

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Lipids

Abstract

Shotgun lipidomics is a well-suited approach to monitor lipid alterations due to its ability to scan for varying lipid types on a global, class and individual species level. However, the ability to perform high-throughput shotgun lipidomics has remained challenging due to time-consuming data processing and hardware limitations. To increase the throughput nature of shotgun lipidomics, an automated shotgun lipidomics approach is described utilizing conventional low flow gradient liquid chromatography (LC) analysis (post-injection) coupled with multiple sample injections per sample (on a lipid scan per injection basis). The proposed automated multi-injection approach resulted in a reproducible lipid scanning period of 2.5 min (in a 4.5 min total data acquisition period), thereby providing a sufficient scanning period for performing either mass spectrometric or tandem mass spectrometric analyses. In addition to being simple, robust and reproducible, this approach was also constructed to be cost-effective by using common LC instrumentation and customizable as the data acquisition period can be tailored to perform different scan types, period lengths and scan numbers. Combined with a strategy to create multiple lipid-specific aliquots per sample, the overall approach provides a simple and efficient platform to perform high-throughput lipid profiling.

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Abbreviations

Ptd2Gro:

Cardiolipins

Cer:

Ceramides

CE:

Cholesteryl esters

DAG:

Diacylglycerols

ESI:

Electrospray ionization

FFA:

Free fatty acids

FIA:

Flow-injection analysis

FS:

Full scan

CerGal:

Galactosylceramides

LC:

Liquid chromatography

LysoPtdCho:

Lysophosphatidylcholines

LysoPtdEtn:

Lysophosphatidylethanolamines

MS:

Mass spectrometry

MRM:

Multiple-reaction Monitoring

NL:

Neutral loss

PtdOH:

Phosphatidic acid

PtdCho:

Phosphatidylcholines

PtdEtn:

Phosphatidylethanolamines

PtdGro:

Phosphatidylglycerols

PtdIns:

Phosphatidylinositols

PtdSer:

Phosphatidylserines

PI:

Precursor ion

CerPCho:

Sphingomyelins

ST:

Sulfatides

MS/MS:

Tandem mass spectrometry

TAG:

Triacylglycerols

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Authors and Affiliations

  1. Hollings Marine Laboratory, National Institute of Standards and Technology, 331 Fort Johnson Road, Charleston, SC, 29412, USA

    John A. Bowden & John R. Kucklick

  2. Department of Obstetrics/Gynecology, Medical University of South Carolina, 96 Jonathan Lucas Street, Suite 634, MSC 619, Charleston, SC, 29425, USA

    Jackie T. Bangma

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  1. John A. Bowden
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  2. Jackie T. Bangma
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Correspondence to John A. Bowden.

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Certain commercial equipment, instruments, or materials are identified in this paper to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology; nor does it imply that the materials or equipment identified are necessarily the best for the purpose.

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Bowden, J.A., Bangma, J.T. & Kucklick, J.R. Development of an Automated Multi-Injection Shotgun Lipidomics Approach Using a Triple Quadrupole Mass Spectrometer. Lipids 49, 609–619 (2014). https://doi.org/10.1007/s11745-014-3903-x

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  • DOI: https://doi.org/10.1007/s11745-014-3903-x

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