Compound-specific Stable Isotope Measurements Research Articles

Turning a 3D Printer into a HPLC Fraction Collector: A Tool for Compound-Specific Stable Isotope Measurements

Compound-specific isotope analysis (CSIA) can provide unique insights into the cycling of elements including carbon and nitrogen. One approach for CSIA is the use of high-performance liquid chromatography (HPLC) to separate compounds of interest, followed by analysis of these compounds using an elemental analyser coupled to an isotope ratio mass spectrometer. A key component of this technique is the fraction collector, which automatically collects compounds as they are separated by HPLC. Here, we present a fraction collector that is a simple adaptation of a 3D printer, and, thus, can be easily adopted by any laboratory already equipped for HPLC. In addition to the much lower cost compared to commercial alternatives, this adaptation has the advantage for CSIA that the 3D printer is able to heat the collected fractions, which is not true for many commercial fraction collectors. Heating allows faster evaporation of the solvent, so that the dried compounds can be measured by EA–IRMS immediately. The procedure can be repeated consecutively so that diluted solutions can have the compounds concentrated for analysis. Any computer-controlled HPLC can be integrated to the fraction collector used here by means of AutoIt.

Advantages of compound‐specific stable isotope measurements over bulk measurements in studies on plant uptake of intact amino acids

Increasing interest in the ability of plants to take up amino acids has given rise to questions on the accuracy of the commonly used bulk method to measure and calculate amino acid uptake. This method uses bulk measurements of 13C and 15N enrichment in plant tissues after application of dual-labelled amino acids but some authors have recommended the use of compound-specific stable isotope (CSI) analysis of the plants' amino acids instead. However, there has never been a direct evaluation of both methods. We conducted a field study applying dual-labelled (13C, 15N) amino acids (glycine, valine, tyrosine and lysine) to soil of a Plantago lanceolata monoculture. Root and shoot samples were collected 24 h after label application and the isotope composition of the plant tissues was investigated using bulk and CSI measurements. Enrichment of 13C in the case of CSI measurements was limited to the applied amino acids, showing that no additional 13C had been incorporated into the plants' amino acid pool via the uptake of tracer-derived C-fragments. Compared with this rather conservative indicator of amino acid uptake, the 13C enrichment of bulk measurements was 8, 5, 1.6 and 6 times higher for fine roots, storage roots, shoot and the whole plant, respectively. These findings show that the additional uptake of tracer-derived C-fragments will result in a considerable overestimation of amino acid uptake in the case of bulk measurements. We therefore highly recommend the use of CSI measurements for future amino acid uptake studies due to their higher accuracy.

Aerobic Biodegradation of Chlorinated Ethenes in a Fractured Bedrock Aquifer: Quantitative Assessment by Compound-Specific Isotope Analysis (CSIA) and Reactive Transport Modeling

A model-based analysis of concentration and isotope data was carried out to assess natural attenuation of chlorinated ethenes in an aerobic fractured bedrock aquifer. Tetrachloroethene (PCE) concentrations decreased downgradient of the source, but constant delta13C signatures indicated the absence of PCE degradation. In contrast, geochemical and isotopic data demonstrated degradation of trichloroethene (TCE) and cis-1,2-dichloroethene (DCE) under the prevailing oxic conditions. Numerical modeling was employed to simulate isotopic enrichment of chlorinated ethenes and to evaluate alternative degradation pathway scenarios. Existing field information on groundwater flow, solute transport, geochemistry, and delta13C signatures of the chlorinated ethenes was integrated via reactive transport simulations. The results provided strong evidence for the occurrence of aerobic TCE and DCE degradation. The chlorinated ethene concentrations together with stable carbon isotope data allowed us to reliably constrain the assessment of the extent of biodegradation at the site and plume simulations quantitatively linked aerobic biodegradation with isotope signatures in the field. Our investigation provides the first quantitative assessment of aerobic biodegradation of chlorinated ethenes in a fractured rock aquifer based on compound specific stable isotope measurements and reactive transport modeling.

Combining hydropyrolysis and compound specific stable isotope measurements to identify sources of biodegrade PAHs in soils and sediments

Combining hydropyrolysis and compound specific stable isotope measurements to identify sources of biodegrade PAHs in soils and sediments

Origin and transport of n-alkane-2-ones in a subtropical estuary: potential biomarkers for seagrass-derived organic matter

n-Alkane-2-ones are lipids commonly found in sediments and soils. This group of compounds, frequently reported in the literature, usually occurs in the form of a homologous series ranging from about C19 to C33 characterized by a strong odd over even carbon number predominance. In this paper we report a different molecular distribution, centered about the C25 homologue as the dominant ketone. The relative abundance of the C25 compared to the C27 homologue in a sediment transect increased from the upper to the lower end of a South Florida estuary, and was found to correlate with surface water salinity in extracts from suspended solids. Analyses of different varieties of seagrasses showed these to be the most likely source of the C25n-alkane-2-ones, while the C27+ homologues were mainly derived from mangroves and freshwater marsh vegetation. Compound-specific stable isotope measurements and statistical analyses support this finding, suggesting that molecular distributions of n-alkane-2-ones can be used to identify seagrass-derived organic matter in coastal environments.

Lipids as carriers of anthropogenic signals from prehistory

Studies performed during the last two decades have shown that lipids are preserved in association with a wide range of artefacts and ecofacts recovered from archaeological sites, e.g. pottery vessels and skeletal remains. The majority of work in this area has focused on the use of molecular structures (‘biomarkers’) and distributions (‘fingerprints’) to assess the nature and origin of commodities associated with past cultural, economic and agricultural practices. However, since lipids, like all other classes of biomolecule, are affected by degradation (both pre– and post–burial), emphasis is now being placed on the complementary use of diagenetically robust, compound–specific stable isotope measurements to enhance the scope and reliability of archaeological interpretations. A feature of theδ13C values of individual lipids, rather than bulk measurements of biochemically more heterogeneous materials, lies in their capacity to reflect differences in both the isotopic composition of the carbon sources used in their biosynthesis and the routing of dietary lipids and their metabolites in consumer organisms. This isotopic information, accessible by gas chromatography–combustion–isotope ratio mass spectrometry, has opened up new avenues of investigation concerning human activity in prehistory.