Measurement of lipogenic flux by deuterium resolved mass spectrometry

Abstract

De novo lipogenesis (DNL) is disrupted in a wide range of human disease. Thus, quantification of DNL may provide insight into mechanisms and guide interventions if it can be performed rapidly and noninvasively. DNL flux is commonly measured by ²H incorporation into fatty acids following deuterated water (²H₂O) administration. However, the sensitivity of this approach is limited by the natural abundance of ¹³C, which masks detection of ²H by mass spectrometry. Here we report that high-resolution Orbitrap gas-chromatography mass-spectrometry resolves ²H and ¹³C fatty acid mass isotopomers, allowing DNL to be quantified using lower ²H₂O doses and shorter experimental periods than previously possible. Serial measurements over 24-hrs in mice detects the nocturnal activation of DNL and matches a ³H-water method in mice with genetic activation of DNL. Most importantly, DNL is detected in overnight-fasted humans in less than an hour and is responsive to feeding during a 4-h study. Thus, ²H specific MS provides the ability to study DNL in settings that are currently impractical.

Fat synthesis is necessary for normal physiology, but its dysregulation contributes to the pathology of many diseases. Here, the authors report a high-resolution mass spectrometry approach that quantifies fat synthesis flux in humans and mice following a brief and low dose of deuterated water.