A framework for the extraction and interpretation of organic molecules in speleothem carbonate

The organic content of speleothem calcite is a well-recognized component of their chemical composition. To date, the techniques for interpretation of this material include UV fluorescence, FTIR spectroscopy and biomarker analysis using gas chromatography/mass spectroscopy (GC/MS). However, investigation of the minute concentrations of molecules in speleothems demands careful sampling and laboratory controls.

To be certain extracted molecules were encapsulated at the time of speleothem growth and do not represent contamination, we submitted three pieces of speleothem calcite to a rigorous extraction procedure. Based on sequential digestion and analysis by GC/MS, we measured concentration profiles of individual compounds with increasing distance from sample surfaces.

Declining concentrations toward interior extracts identified cholesterol, phthalates, and n-alkanes as surface contaminants. In contrast, iodo organic compounds had homogeneous concentration profiles and were also significantly above laboratory background levels, consistent with an indigenous origin. However, further laboratory testing demonstrated that iodo organics were produced by the reaction of iodine derived from the speleothem with solvent additives and other impurities of the extraction procedure. Sitosterol and some fatty acids demonstrated distributions which were probably indigenous to the speleothem archive, thus recording environmental conditions commensurate with time of growth.

We do not aim to provide an environmental interpretation of extracted molecules, but highlight the caution necessary before doing so. We ultimately establish a framework for differentiating between organic constituents that are introduced to the speleothems during storage, handling and as artifacts of extraction, and those encapsulated in situ at the time of growth.