Skip to main content
SpringerLink
Log in

Simultaneous Identification and Quantification of Triacyglycerol Species in Human Plasma by Flow-Injection Electrospray Ionization Tandem Mass Spectrometry

  • Original
  • Published:
Chromatographia Aims and scope Submit manuscript

Abstract

The analysis of triacylglycerol (TG) species is of interest in diseases that are associated with hypertriglycidemia. The individual composition of TGs seems to be of special interest in the development of atherosclerosis or diabetes type II. Enzymatic methods based on saponification and glycerol analysis are not suited for the TG fatty acid distribution. The aim of the study was to develop a rapid method for a molecular species fingerprinting of TGs. Protein precipitation of 5 µL human plasma was carried out with toluene/methanol (1:1 v/v). Tandem mass spectrometric detection (mass range m/z 700–1,000) was performed by combination of nine neutral loss scans (14:0, 15:0, 16:0, 16:1, 18:0, 18:1, 18:2, 18:3, and 20:4) after flow-injection analysis for 2 min. Deuterated internal standards had been used for quantification. In human EDTA-plasma the detection limit was 3.3 µg/mL and the lower limit of quantification was 11.1 µg/mL. Linearity was proved for TG concentrations up to 100 µg/mL for each TG species. Nineteen TG molecular species were determined with an intra-day coefficient of variation of 15.0–20.9 % (n = 9), and an inter-day coefficient of variation of 17.3–36.6 % (n = 9). Recovery of TG 50:0 using the equivalent internal standard was 97.0 %. Nineteen TG molecular species can be analyzed in 2 min from human plasma or serum by the novel tandem mass spectrometric approach. In subsequent studies, the distribution of plasma TG molecular species can be analyzed under high-throughput conditions in healthy and diseased individuals.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Storlien LH, Kriketos AD, Calvert GD, Baur LA, Jenkins AB (1997) Fatty acids, triglycerides and syndromes of insulin resistance. Prostaglandins Leukot Essent Fatty Acids 57(4–5):379–385. doi:10.1016/S0952-3278(97)90414-2

    Article  CAS  Google Scholar 

  2. Nordestgaard BG, Benn M, Schnohr P, Tybjærg-Hansen A (2007) Nonfasting triglycerides and risk of myocardial infarction, ischemic heart disease, and death in men and women. JAMA 298(3):299–308

    Article  CAS  Google Scholar 

  3. Han X, Gross RW (2001) Quantitative Analysis and Molecular Species Fingerprinting of Triacylglyceride Molecular Species Directly from Lipid Extracts of Biological Samples by Electrospray Ionization Tandem Mass Spectrometry. Anal Biochem 295(1):88–100. doi:10.1006/abio.2001.5178

    Article  CAS  Google Scholar 

  4. Freiberg JJ, Tybjærg-Hansen A, Jensen J, Nordestgaard BG (2008) NOnfasting triglycerides and risk of ischemic stroke in the general population. JAMA 300(18):2142–2152

    Article  CAS  Google Scholar 

  5. Rhee EP, Cheng S, Larson MG, Walford GA, Lewis GD, McCabe E, Yang E, Farrell L, Fox CS, O’Donnell CJ, Carr SA, Vasan RS, Florez JC, Clish CB, Wang TJ, Gerszten RE (2011) Lipid profiling identifies a triacylglycerol signature of insulin resistance and improves diabetes prediction in humans. J Clin Investig 121(4):1402–1411. doi:10.1172/jci44442

    Article  CAS  Google Scholar 

  6. McBride PE (2007) Triglycerides and risk for coronary heart disease. JAMA 298(3):336–338

    Article  CAS  Google Scholar 

  7. Hunter JE (2001) Studies on effects of dietary fatty acids as related to their position on triglycerides. Lipids 36(7):655–668. doi:10.1007/s11745-001-0770-0

    Article  CAS  Google Scholar 

  8. Krank J, Murphy RC, Barkley RM, Duchoslav E, McAnoy A (2007) Qualitative analysis and quantitative assessment of changes in neutral glycerol lipid molecular species within cells. In: Brown HA (ed) Methods in enzymology, vol 432. Academic Press, USA, pp 1–20. doi:10.1016/S0076-6879(07)32001-6

    Google Scholar 

  9. Fernandez C, Sandin M, Sampaio JL, Almgren P, Narkiewicz K, Hoffmann M, Hedner T, Wahlstrand B, Simons K, Shevchenko A, James P, Melander O (2013) Plasma Lipid Composition and Risk of Developing Cardiovascular Disease. PLoS ONE 8(8):e71846

    Article  CAS  Google Scholar 

  10. Schwudke D, Oegema J, Burton L, Entchev E, Hannich JT, Ejsing CS, Kurzchalia T, Shevchenko A (2005) Lipid Profiling by Multiple Precursor and Neutral Loss Scanning Driven by the Data-Dependent Acquisition. Anal Chem 78(2):585–595. doi:10.1021/ac051605m

    Article  Google Scholar 

  11. Hsu F-F, Turk J (2010) Electrospray ionization multiple-stage linear ion-trap mass spectrometry for structural elucidation of triacylglycerols: assignment of fatty acyl groups on the glycerol backbone and location of double bonds. J Am Soc Mass Spectrom 21(4):657–669. doi:10.1016/j.jasms.2010.01.007

    Article  CAS  Google Scholar 

  12. Myher JJ, Kuksis A, Marai L, Sandra P (1988) Identification of the more complex triacylglycerols in bovine milk fat by gas chromatography—mass spectrometry using polar capillary columns. J Chromatogr A 452:93–118. doi:10.1016/S0021-9673(01)81440-0

    Article  CAS  Google Scholar 

  13. Hoving EB, Jansen G, Volmer M, van Doormaal JJ, Muskiet FAJ (1988) Profiling of plasma cholesterol ester and triglyceride fatty acids as their methyl esters by capillary gas chromatography, preceded by a rapid aminopropyl-silica column chromatographic separation of lipid classes. J Chromatogr B Biomed Sci Appl 434(2):395–409. doi:10.1016/S0378-4347(88)80006-9

    Article  CAS  Google Scholar 

  14. McAnoy AM, Wu CC, Murphy RC (2005) Direct qualitative analysis of triacylglycerols by electrospray mass spectrometry using a linear ion trap. J Am Soc Mass Spectrom 16(9):1498–1509. doi:10.1016/j.jasms.2005.04.017

    Article  CAS  Google Scholar 

  15. Bligh EG, Dyer WJ (1959) A rapid method for total lipid extraction and purification. Can J Biochem Physiol 37:911–917

    Article  CAS  Google Scholar 

  16. Folch J, Lees M, Stanley GHS (1957) A simple method for the isolation and purification of total lipides from animal tissue. J Biol Chem 226:497–509

    CAS  Google Scholar 

  17. http://webbook.nist.gov. Accessed 15 April 2014

  18. Fliszar KA, Peter Wuelfing W, Li Z, Reed RA (2006) Profiling of medium chain glycerides used in pharmaceutical formulation development by reversed-phase HPLC. J Pharm Biomed Anal 40(4):896–900. doi:10.1016/j.jpba.2005.08.024

    Article  CAS  Google Scholar 

  19. Holčapek M, Dvořáková H, Lísa M, Girón AJ, Sandra P, Cvačka J (2010) Regioisomeric analysis of triacylglycerols using silver-ion liquid chromatography–atmospheric pressure chemical ionization mass spectrometry: Comparison of five different mass analyzers. J Chromatogr A 1217(52):8186–8194. doi:10.1016/j.chroma.2010.10.064

    Article  Google Scholar 

  20. Dillon JT, Aponte JC, Tarozo R, Huang Y (2012) Efficient liquid chromatographic analysis of mono-, di-, and triglycerols using silver thiolate stationary phase. J Chromatogr A 1240:90–95. doi:10.1016/j.chroma.2012.03.083

    Article  CAS  Google Scholar 

  21. Leiker TJ, Barkley RM, Murphy RC (2011) Analysis of diacylglycerol molecular species in cellular lipid extracts by normal-phase LC-electrospray mass spectrometry. Int J Mass Spectrom 305(2–3):103–108. doi:10.1016/j.ijms.2010.09.008

    Article  CAS  Google Scholar 

  22. Hutchins PM, Barkley RM, Murphy RC (2008) Separation of cellular nonpolar neutral lipids by normal-phase chromatography and analysis by electrospray ionization mass spectrometry. J Lipid Res 49(4):804–813. doi:10.1194/jlr.M700521-JLR200

    Article  CAS  Google Scholar 

  23. Nagy K, Sandoz L, Destaillats F, Schafer O (2012) Mapping the regioisomeric distribution of fatty acids in triacylglycerols by hybrid mass spectrometry. J Lipid Res. doi:10.1194/jlr.D031484

    Google Scholar 

  24. Nagy K, Sandoz L, Destaillats F, Schafer O (2013) Mapping the regioisomeric distribution of fatty acids in triacylglycerols by hybrid mass spectrometry. J Lipid Res 54(1):290–305. doi:10.1194/jlr.D031484

    Article  CAS  Google Scholar 

  25. Lembcke J, Ceglarek U, Fiedler GM, Baumann S, Leichtle A, Thiery J (2005) Rapid quantification of free and esterified phytosterols in human serum using APPI-LC-MS/MS. J Lipid Res 46(1):21–26. doi:10.1194/jlr.C400004-JLR200

    Article  CAS  Google Scholar 

  26. Quehenberger O, Armando AM, Brown AH, Milne SB, Myers DS, Merrill AH, Bandyopadhyay S, Jones KN, Kelly S, Shaner RL, Sullards CM, Wang E, Murphy RC, Barkley RM, Leiker TJ, Raetz CRH, Guan Z, Laird GM, Six DA, Russell DW, McDonald JG, Subramaniam S, Fahy E, Dennis EA (2010) Lipidomics reveals a remarkable diversity of lipids in human plasma. J Lipid Res 51(11):3299–3305. doi:10.1194/jlr.M009449

    Article  CAS  Google Scholar 

  27. Roche_Diagnostics_GmbH (2011) Methodenbeschreibung Triglycerides für cobas c System

  28. Liu L, Zhang Y, Chen N, Shi X, Tsang B, Yu Y-H (2007) Upregulation of myocellular DGAT1 augments triglyceride synthesis in skeletal muscle and protects against fat-induced insulin resistance. J Clin Investig 117(6):1679–1689. doi:10.1172/jci30565

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Liebigstr. 27, 04103, Leipzig, Germany

    M. Sander, S. Becker, J. Thiery & U. Ceglarek

  2. LIFE–Leipzig Research Center for Civilization Diseases, University Leipzig, Leipzig, Germany

    S. Becker, J. Thiery & U. Ceglarek

Authors
  1. M. Sander
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Thiery
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. U. Ceglarek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. Sander.

Additional information

Published in the topical collection Recent Developments in Clinical Omics with guest editors Martin Giera and Manfred Wuhrer.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 172 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sander, M., Becker, S., Thiery, J. et al. Simultaneous Identification and Quantification of Triacyglycerol Species in Human Plasma by Flow-Injection Electrospray Ionization Tandem Mass Spectrometry. Chromatographia 78, 435–443 (2015). https://doi.org/10.1007/s10337-014-2782-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10337-014-2782-x

Keywords

Search

Navigation