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FAME TG preprint.pdf (500.2 kB)

Fatty Acid Sentinels as Covalently Bound Randomization Standards for Triacylglycerol (TAG) Quantitative Analysis

preprint
submitted on 02.07.2020 and posted on 06.07.2020 by DongHao Wang, Zhen Wang, Secilia Martinez, Herbert Tobias, Tom Brenna
RATIONALE. Quantitative analysis of triacylglycerols (TAG) is impeded by a lack of standards and the huge number of potential TAG molecular species that may be present due to the combinatorial nature of glycerolipids. Randomization of acyl groups yields TAG mixtures with profiles predictable from fatty acid profiles, however their use as calibration mixtures has been limited. METHODS. We introduce here the principle of fatty acid isotopic sentinels that are quantitatively added prior to randomization to enable verification that randomization is complete, and that can be used as internal standards. A mixture of two or more
isotopically labeled fatty acid methyl esters (FAME) are prepared in quantitative proportions and randomized covalently into the acyl groups of TAG mixtures.
RESULTS. Reaction with catalytic amounts of NaOCH3 yields complete randomization, such that the product FAME and TAG have the same fatty acid profile. TAG mixture
analysis reveals that the isotopic sentinels have been covalently incorporated into TAG molecular species and <1% of the expected proportions thus verifying randomization within experimental error.
CONCLUSIONS. The sentinel principle demonstrated here as covalently incorporated internal standards verifies that randomization chemistry went to completion. It applies in
general to use of combinatorial chemistry for quantitative standards.

History

Email Address of Submitting Author

tbrenna@utexas.edu

Institution

University of Texas at Austin

Country

United States

ORCID For Submitting Author

0000-0001-9494-4245

Declaration of Conflict of Interest

None

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