Use of [13C18] Oleic Acid and Mass Isotopomer Distribution Analysis to Study Synthesis of Plasma Triglycerides In Vivo: Analytical and Experimental Considerations
2013
McLaren, David G. | Cardasis, Helene L. | Stout, Steven J. | Wang, Shengbing | Mendoza, Vivienne | Castro-Perez, Jose M. | Miller, Paul L. | Murphy, Beth A. | Cumiskey, Anne-Marie | Cleary, Michele A. | Johns, Douglas G. | Previs, Stephen F. | Roddy, Thomas P.
We have previously reported on a liquid chromatography–mass spectrometry method to determine the disposition of [¹³C₁₈]-oleic acid following intravenous and oral administration in vivo. This approach has enabled us to study a variety of aspects of lipid metabolism including a quantitative assessment of triglyceride synthesis. Here we present a more rigorous evaluation of the constraints imposed upon the analytical method in order to generate accurate data using this stable-isotope tracer approach along with more detail on relevant analytical figures of merit including limits of quantitation, precision, and accuracy. The use of mass isotopomer distribution analysis (MIDA) to quantify plasma triglyceride synthesis is specifically highlighted, and a re-evaluation of the underlying mathematics has enabled us to present a simplified series of equations. The derivation of this MIDA model and the significance of all underlying assumptions are explored in detail, and examples are given of how it can successfully be applied to detect differences in plasma triglyceride synthesis in lean and high-fat diet fed mouse models. More work is necessary to evaluate the applicability of this approach to triglyceride stores with slower rates of turnover such as in adipose or muscle tissue; however, the present report provides investigators with the tools necessary to conduct such studies.
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