We examined the potential of stable-isotope Raman microspectroscopy (SIRM) for the evaluation of differently enriched 13C-labeled humic acids as model substances for soil organic matter (SOM). The SOM itself can be linked to the soil water holding capacity. Therefore, artificial humic acids (HA) with known isotopic compositions were synthesized and analyzed by means of SIRM. By performing a pregraphitization, a suitable analysis method was developed to cope with the high fluorescence background. Results were verified against isotope ratio mass spectrometry (IRMS). The limit of quantification was 2.1 × 10-1 13C/C tot for the total region and 3.2 × 10-2 13C/C tot for a linear correlation up to 0.25 13C/C tot. Complementary nanoscale secondary ion mass spectrometry (NanoSIMS) analysis indicated small-scale heterogeneity within the dry sample material, even though-owing to sample topography and occurring matrix effects-obtained values deviated in magnitude from those of IRMS and SIRM. Our study shows that SIRM is well-suited for the analysis of stable isotope-labeled HA. This method requires no specific sample preparation and can provide information with a spatial resolution in the micrometer range. Graphical abstract Analysis of the isotopic composition of humic acids by Raman microspectroscopy in combination with isotope ratio mass spectrometry and nanoscale secondary ion mass spectrometry.
Keywords: Humic acids; Raman microspectroscopy; Soil organic matter; Stable isotopes.