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. 2023 Jul 8:11:102283.
doi: 10.1016/j.mex.2023.102283. eCollection 2023 Dec.

Protocols for sample preparation and compound-specific stable-isotope analyses (δ2H, δ13C) of fatty acids in biological and environmental samples

Matthias Pilecky  1   2 , Leonard I Wassenaar  1   2 , Sami Taipale  3 , Martin J Kainz  1   2
Affiliations

Protocols for sample preparation and compound-specific stable-isotope analyses (δ2H, δ13C) of fatty acids in biological and environmental samples

Matthias Pilecky et al. MethodsX. .

Abstract

Compound-specific stable-isotope analysis (CSIA) of fatty acids is a powerful tool to better understand the trophic transfer of fatty acids and their biochemical fate in and across ecosystems, including tracing animal migration and understanding physiological processes. The non-exchangeable nature of C-H bonds in acyl chains, hydrogen (δ2H) and carbon (δ13C) stable-isotope values of fatty acids (FA) provide independent information about the origins of fatty acids. Several technical obstacles must be overcome to ensure accurate and reproducible measurements of FA-CSIA can be made. This protocol describes the sample preparation process for successful stable-isotope analyses of fatty acids obtained from environmental and biological samples. Numerous techniques for the preanalytical processing of fatty acid samples are available, and these often have minimal impact on δ values. Here, we provide an in-depth guide detailing our well-established laboratory protocols, ranging from the initial sample preparation, lipid extraction, and transmethylation to the instrumental arrangement, data collection, and analysis.•Protocol from obtaining a sample to standardized fatty acid specific δ2H and δ13C values.•Separate GC analysis procedures for C and H are recommended for optimal performance.

Keywords: CSIA; Carbon isotopes; Deuterium; Fatty acids; GC-IRMS; Reproducible determinations of the δ2H and δ13C composition of fatty acids from environmental samples.; Stable-isotopes.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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