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Direct Derivatization vs Aqueous Extraction Methods of Fecal Free Fatty Acids for GC–MS Analysis

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Lipids

Abstract

A comprehensive and accurate determination of free fatty acids (FFA) is required for fecal metabolomic investigations. The present study compares three aqueous extraction methods (1) ULTRA-TURRAX®, (2) whirl mixing and (3) basic ULTRA-TURRAX extraction of fecal FFA with a direct derivatization approach using ethyl chloroformate as the derivatization reagent before determination by gas chromatography–mass spectrometry. The direct derivatization method resulted in significantly higher estimations (P < 0.01) of short- and long-chain fatty acids than was the case when applying the aqueous extraction methods using ULTRA-TURRAX, whirl mixing, or basic ULTRA-TURRAX extraction before the derivatization step. Thus, avoiding an aqueous extraction before derivatization reduces the loss of volatile short-chain FFA and the less water-soluble long-chain FFA.

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Abbreviations

BCFAA:

Branched chain free amino acids

ECF:

Ethyl chloroformate

FFA:

Free fatty acids

GC:

Gas chromatography

GC–MS:

Gas chromatography–mass spectrometry

LCFFA:

Long chain free fatty acids

RSD:

Relative standard deviation

SCFFA:

Short chain free fatty acids

SIM:

Single ion monitoring

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Acknowledgments

The Danish Council for Strategic Research, the Danish Dairy Research Foundation, and Arla Foods are acknowledged for their financial support through the two projects ‘DairyHealth - Dairy lipids, proteins and the metabolic syndrome’ (Journal Number 10-093497) and ‘FIAF - Milk in regulating lipid metabolism and overweight. Uncovering milk’s ability to increase expression and activity of fasting-induced adipose factor’ (Journal Number 10-093539).

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Authors and Affiliations

  1. Department of Food Science, Aarhus University, Blichers Allé 20, Tjele, 8830, Denmark

    Bashar Amer, Caroline Nebel & Trine K. Dalsgaard

  2. Department of Food Science, Aarhus University, Kirstinebjergvej 10, Årslev, 5792, Denmark

    Hanne C. Bertram

  3. Arla Strategic Innovation Centre, Rørdrumvej 2, Brabrand, 8220, Denmark

    Grith Mortensen

Authors
  1. Bashar Amer
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  2. Caroline Nebel
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  3. Hanne C. Bertram
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  4. Grith Mortensen
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  5. Trine K. Dalsgaard
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Correspondence to Trine K. Dalsgaard.

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Amer, B., Nebel, C., Bertram, H.C. et al. Direct Derivatization vs Aqueous Extraction Methods of Fecal Free Fatty Acids for GC–MS Analysis. Lipids 50, 681–689 (2015). https://doi.org/10.1007/s11745-015-4029-5

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  • DOI: https://doi.org/10.1007/s11745-015-4029-5

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