Abstract
Careful assessment of fractionation associated with High Performance Liquid Chromatography (HPLC) is essential for ensuring accurate carbon isotope ratio (δ13C) measurements. Here we report these effects for orthogonal dimensions of HPLC separation of iGDGTs. We observe 13C fractionation during normal phase separations using an amino-bonded stationary phase, finding that the heavy isotope preferentially elutes relative to its lighter counterpart, while negligible fractionation occurs during reverse phase HPLC. We argue that the fractionation during normal phase is caused by diminished Van der Waals attraction between 13C-enriched iGDGTs and the stationary phase and appears to depend on the structure of the analyte. Ultimately, these results emphasize the necessity of complete peak collection to preserve the isotopic character of iGDGTs.
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