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Determination of the fatty acid profile of neutral lipids, free fatty acids and phospholipids in human plasma

  • Nina Firl , Hermine Kienberger , Teresa Hauser and Michael Rychlik EMAIL logo
Published/Copyright: September 25, 2012
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 51 Issue 4

Abstract

Background: Knowledge of the fatty acid composition of lipid classes in human plasma is an important factor in the investigation of human metabolism. Therefore, a method for the analysis of neutral lipid (NL), phospholipid (PL) and free fatty acids (FFA) in human plasma has been developed and validated.

Methods: Separation of lipid classes was carried out by solid phase extraction of the lipid extract. The fractions were transesterified and the resulting fatty acid methyl esters were determined by GC/FID. For the method to be validated, precision, detection and quantification limits, as well as recovery, were determined for combined lipid extraction, solid phase extraction and GC analysis.

Results: The lipid extraction was miniaturized and simplified by application of an ultrasound ‘Sonotrode’. The resolution of lipid classes was optimized with appropriate standards added to a representative plasma sample. In addition, a rapid derivatization procedure using trimethylsulfoniumhydroxide was established. Low determination limits (1.5, 0.2 and 1.3 μg/g plasma for NL, PL and FFA, respectively) indicate that the method’s sensitivity is sufficient to quantify even minor components. Furthermore, recovery for NL and PL fatty acids was found to range from 80% to 110%. The results were similar for FFA apart from more polar free fatty acids due to their higher solubility in water. Repetitive measurements showed very good precision apart from the long chain PUFA for which the coefficients of variation were significantly higher.

Conclusions: The present method is applicable to the quantitation of fatty acids in lipid classes of human plasma including several minor components.

Keywords: fatty acids; gas chromatography; lipid separation; miniaturized ultrasonication; plasma lipids, solid phase extraction; TMSH; validation
Corresponding author: Michael Rychlik, Chair of Analytical Food Chemistry, Technische Universität München, Alte Akademie 10, 85354 Freising, Germany, Phone: +49 8161 713153, Fax: +49 8161 714216

We thank Susanne Grill, Technische Universität München, for assisting in the determination of endogenous fatty acids and Dr. Thomas Skurk, Chair for Nutritional Medicine, Technische Universität München, for taking blood samples from the volunteers. Moreover, the authors gratefully acknowledge the support of the Faculty Graduate Center Weihenstephan of TUM Graduate School at Technische Universität München, Germany. We thankfully appreciate Thea Varvis’s assistance in language editing the manuscript.

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

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Received: 2012-03-29
Accepted: 2012-08-19
Published Online: 2012-09-25
Published in Print: 2013-04-01

©2013 by Walter de Gruyter Berlin Boston

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https://doi.org/10.1515/cclm-2012-0203
Keywords for this article
fatty acids; gas chromatography; lipid separation; miniaturized ultrasonication; plasma lipids, solid phase extraction; TMSH; validation

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  2. Missing agreement between the two IMMULITE® PSA assays
  3. “Cerebrovascular stressing”: dipyridamole-induced S100B elevation predicts ischemic cerebrovascular events
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  22. Opinion Papers
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  25. Guidelines and Recommendations
  26. A position paper of the EFLM Committee on Education and Training and Working Group on Distance Education Programmes/E-Learning: developing an e-learning platform for the education of stakeholders in laboratory medicine
  27. General Clinical Chemistry and Laboratory Medicine
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