Search Results (22)

This study evaluated the authenticity of olive oil in canned tuna products from the Slovenian market using both official methods, including fatty acid (FA) profiling, determination of the equivalent carbon number difference (ΔECN42), and sterol analysis, and an advanced method: stable carbon isotope analysis (δ¹³C) of FAs obtained through compound-specific isotope analysis (CSIA). Results from both methods confirmed that all 10 samples were authentic, as per the limits set by EU Regulation 2022/2104 and supported by the scientific literature. Method performance was further evaluated by adulterating the olive oil from the canned tuna with 5–20% vegetable oil (VO) or hazelnut oil (HO). While FA analysis struggled to differentiate adulterants with similar FA profiles, CSIA of FAs significantly improved detection. However, distinguishing between VO and HO blended samples remained challenging. PLS-DA analysis further supported the potential of using δ¹³C values of FA for food authentication. Storage of adulterated samples also influenced FA composition, leading to significant changes in MUFA/PUFA ratios and δ¹³C values, which became less negative, likely due to oxidative degradation. In summary, the combination of official and advanced methods, supported by chemometric analysis, offers a robust approach to ensuring the authenticity of olive oil in canned tuna. Full article

Isotopic analyses are useful tools with a wide range of applications, including environmental studies, archaeology and biomedicine. Founded in 2019 at the University of Campania “Luigi Vanvitelli”, the iCONa laboratory specialises in stable isotope mass spectrometry, with a particular focus on cultural heritage. The laboratory performs carbon, nitrogen and oxygen isotopic analyses, including the most recent advances in compound-specific stable isotope analysis of amino acids (CSIA-AAs). In addition to these analytical services, iCONa provides chemical and physical sample preparation for a variety of sample types. This paper focuses on our applications in the field of cultural heritage, exploring how the analysis of stable isotopes performed on archaeological remains can be used to reconstruct past subsistence strategies and human behaviours. We also discuss the challenges inherent in isotopic analysis and recent methodological advances in the field. Full article

Stable isotope analysis is a powerful tool for inferring and quantifying transformation processes, but its effectiveness relies on understanding the magnitude and variability of isotopic fractionation associated with specific reactions. Potassium permanganate (KMnO₄) is widely used as an efficient oxidant for the degradation of trichloroethylene (TCE); however, the influence of environmental factors on the isotope fractionation during this process remains unclear. In this study, compound-specific isotope analysis (CSIA) was conducted to investigate the variability in carbon isotope effects during the KMnO₄-mediated degradation of TCE under varying conditions, including initial concentrations of KMnO₄ and TCE, the presence of humic acid (HA), pH levels, and inorganic ions. The results showed that the overall carbon isotope enrichment factors (ε) of TCE ranged from −26.5 ± 0.5‰ to −22.8 ± 0.9‰, indicating relatively small variations across conditions. At low KMnO₄/TCE molar ratio (n(KMnO₄)/n(TCE)), incomplete oxidation and/or MnO₂-mediated oxidation of TCE likely resulted in smaller ε. For dense, non-aqueous phase liquid (DNAPL) TCE, which represents extremely high concentrations, the ε value was −13.0 ± 1.7‰ during KMnO₄ oxidation. This may be attributed to the slow dissolution of isotopically light TCE from the DNAPL phase, altering the δ¹³C signature of the reacted TCE and resulting in a significantly larger ε value than observed for dissolved-phase TCE oxidation. The ε values increased with rising pH, probably due to the decrease in oxidation potential (E₀) of KMnO₄ from pH ~2 to ~12, as well as the emergence of different degradation pathways and intermediates under varying pH conditions. Both SO₄²⁻ and NO₃⁻ slightly influenced the ε values, potentially due to the formation of H₂SO₄ and HNO₃ at lower pH, which may act as auxiliary oxidants and contribute to TCE degradation. A high concentration (50 mM) of HA led to a decrease in ε values, likely due to competitive interactions between HA and TCE for KMnO₄, which reduced the effective oxidation of TCE. Overall, the carbon isotope enrichment factors for KMnO₄-mediated TCE degradation are relatively stable, although certain environmental conditions can exert minor influences. These findings highlight the need for caution when applying quantitative assessment based on CSIA for KMnO₄ oxidation of TCE. Full article

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. Full article

To bridge the gap between lab-scale microcosm research and field application in the compound-specific stable isotope analysis (CSIA) of atrazine, we studied the characteristics of carbon and nitrogen isotope fractionation in the atrazine degradation processes within a constructed wetland. In the wetland, we observed multiple element (C, N) isotope fractionation parameters, such as kinetic isotope effects and dual isotope slopes. These parameters are very consistent with those observed in the cultivation of AtzA- or TrzN-harboring strains, suggesting a similarity in the pathway and reaction mechanism of atrazine biodegradation between the two settings. However, we recorded variable carbon (${\epsilon }_C:$−3.2 ± 0.6‰ to −4.3 ± 0.6‰) and nitrogen isotope fractionation (${\epsilon }_N:$1.0 ± 0.3‰ to 2.2 ± 0.3‰) across different phases. This variance could lead to an over- or underestimation of the biodegradation extent of atrazine when employing the large or small enrichment factor of the carbon isotope. Intriguingly, the estimation accuracy improved considerably when using the enrichment factor (−4.6‰) derived from the batch cultivation of the pore water. This study advances the application of CSIA in tracking atrazine biodegradation processes in ecosystems, and it also underlines the importance of the careful selection and application of the enrichment factor in quantifying the intrinsic biodegradation of atrazine in ecosystems. Full article

Compound-specific stable isotope analysis (CSIA) is an efficient method for source apportionment and the identification of the transformation process in organic compounds. However, most studies of CSIA are still limited to laboratory experiments. Few studies used have CSIA in an in situ environment due to the complexity of environmental samples. Therefore, a purification method for analyzing the carbon isotope ratios of three phenolic endocrine disrupting compounds (EDCs) (nonylphenols (NPs), octylphenol (OP), and bisphenol A(BPA)) in sediment and water samples was developed in this study. The silica gel column was used to isolate EDCs from complex matrices with multiple organic solvents. Gas chromatography/mass spectrometry was used to quantify the targeted EDCs and analyze the purity of the extracts in full-scan mode. The interfering peaks disappeared, the baseline was sharply reduced, and all the target compounds appeared as single peaks in the chromatogram after purification. Analyzing the standard samples with known isotope ratios showed that the purification treatment did not cause isotope fractionation. The isotopic difference before and after purification was less than 0.04. The method was successfully used to analyze the isotope composition of BPA, OP, and NPs in river water and sediments in the Guangzhou River, Pearl River Delta, South China. Sewage discharge significantly affected the carbon isotope values of BPA, OP and NPs in Guangzhou rivers, suggesting that sewage discharge is the main source of EDCs in the Guangzhou rivers. There is a significant correlation between the isotopic values and concentrations of OP and NPs in sediments, indicating that they may undergo chemical transformation. Full article

Compound-specific isotopic analysis (CSIA) and geochemical modeling were applied to evaluate the effectiveness of an 800 m-long sequential in situ bioremediation (ISB) system in Northern Italy. The system was created for the clean-up of a polluted aquifer affected by chloroethenes. A hydraulically upgradient anaerobic (AN)-biobarrier-stimulated reductive dichlorination (RD) of higher chloroethenes (PCE, TCE) and a downgradient aerobic (AE)-biobarrier-stimulated oxidation (OX) of lower chloroethenes (DCE, VC) were proposed. Carbon CSIA and concentration data were collected for PCE, TCE, cis-DCE and VC and interpreted using a reactive transport model that was able to simulate isotopic fractionation. The analysis suggested that the combination of CSIA and modeling was critical to evaluate the efficiency of sequential ISBs for the remediation of chloroethenes. It was found that the sequential ISB could reduce the PCE, TCE and cis-DCE concentrations by >99% and VC concentrations by >84% along the flow path. First-order RD degradation rate constants ($k_{RD}$) increased by 30 times (from $k_{RD}$= 0.2–0.3 y⁻¹ up to $k_{RD}$ = 6.5 y⁻¹) downgradient of the AN barrier. For cis-DCE and VC, the AE barrier had a fundamental role to enhance OX. First-order OX degradation rate constants ($k_{OX}$) ranged between $k_{OX}$= 0.7–155 y⁻¹ for cis-DCE and $k_{OX }$= 1.7–12.6 y⁻¹ for VC. Full article

A new pyrolysis technology involving chromium is proposed for the determination of δD in alkanes based on the systematic analysis of reaction temperature, conversion rate, and reaction mechanism. Compared with the traditional high-temperature conversion (HTC) method, our findings suggest that chromium/high-temperature conversion (Cr/HTC) can improve the conversion rate of hydrocarbons and reduce the required pyrolysis temperature by up to 175 °C; meanwhile, the pyrolysis conversion rate of hydrocarbons increased by an average of 2.42% across the entire analyzed temperature range using the Cr/HTC method. Changes in the chromium wire itself were analyzed using X-ray photoelectron spectroscopy (XPS); this facilitated an understanding of the interaction mechanism between chromium and hydrocarbons and possible pathways of the catalytic pyrolysis process. The results show that chromium reacts with hydrocarbons, capturing carbon as chromium carbide (Cr₂C₃ and CrC₃) and releasing hydrogen in the form of H₂. As the reaction progresses, the resulting free carbon accumulates on the surface of the chromium wire or chromium carbide, resulting in a marked reduction in the Cr/C ratio; these findings provide reliable evidence for the further application of Cr/HTC technology. Full article

Deep-water dogfish (sharks) are caught on Australia’s continental shelf as by-products to other deep-water species with revenue derived from fillets for human consumption and from the livers which are sold for their oil content. Deep-water dogfish utilise a large oil-rich liver for buoyancy, which may account for 20–25% of their body weight. An important constituent of certain dogfish liver oil is squalene, a highly unsaturated triterpenoid (C₃₀H₅₀) hydrocarbon which in some species can be up to 90% of the oil, though in the Australian commercial species it is typically around 50%. Squalene (and deep-water dogfish liver oil in general) has a long-standing high value in products, such as cosmetics and nutraceuticals. Manufacturers are increasingly required to demonstrate the sustainability of products, and this is integral to the importance of demonstrating product provenance. Australia’s mid-slope deep-water dogfish fishery is recognised globally as well managed and sustainable; therefore, it is important to be able to distinguish products derived from these regions from other unregulated, unsustainable and cheaper sources in order to protect Australia’s competitive advantages and ensure sustainability. In this study, we have sourced deep-water dogfish liver oil samples originating from Southeast Australia, New Zealand, India, Northeast Africa and the Arabian Sea. The squalene was isolated by commercial or laboratory processing. A compound-specific carbon stable isotope analysis of the derived squalene was then used to determine isotopic resolution and assign the likely origins of a variety of commonly available off-the-shelf nutraceuticals in Australian outlets. Squalene sourced and produced from Southeast Australian and New Zealand dogfish liver oils showed δ¹³C values in the range of −22.1 to −24.2‰, with all other squalene samples distinguishable at −19.9 to −20.7‰. Many of the off-the-shelf squalene products claiming to be of Australian origin showed δ¹³C values very distinct from the range of the genuine Southeast Australian- and New Zealand-produced squalene. Full article

By using compound-specific isotope analysis (CSIA) in combination with high-throughput sequencing analysis (HTS), we successfully evaluated the benzene and toluene biodegradation in a bio-permeable reactive barrier (bio-PRB) and the stress response of the microbial community. Under stress conditions, a greater decline in the biodegradation rate of BTEX was observed compared with the apparent removal rate. Both an increase in the influent concentration and the addition of trichloroethylene (TCE) inhibited benzene biodegradation, while toluene biodegradation was inhibited by TCE. Regarding the stress response, the relative abundance of the dominant bacterial community responsible for the biodegradation of BTEX increased with the influent concentration. However, the dominant bacterial community did not change, and its relative abundance was restored after the influent concentration decreased. On the contrary, the addition of TCE significantly changed the bacterial community, with Aminicenantes becoming the dominant phyla for co-metabolizing TCE and BTEX. Thus, TCE had a more significant influence on the bio-PRB than an increasing influent concentration, although these two stress conditions showed a similar degree of influence on the apparent removal rate of benzene and toluene. The present work not only provides a new method for accurately evaluating the biodegradation performance and microbial community in a bio-PRB, but also expands the application of compound-specific isotope analysis in the biological treatment of wastewater. Full article

Compound specific isotope analysis (CSIA) and enantiomer specific isotope analysis (ESIA) are powerful tools for assessing the fate of hexachlorocyclohexanes (HCHs) in the environment. However, there is no systematic study on the CSIA and ESIA analysis test methods of the carbon isotopes of HCHs in water and soil environments, in particular the isotope fractionation in the pre-concentration process. We endeavored to test the compatibility of CSIA and ESIA with the liquid–liquid extraction method of HCHs in water. The results showed that there were negligible changes in the δ¹³C of HCHs after extraction, indicating that liquid–liquid extraction can be used as a pre-concentration method for the determination of δ¹³C of HCHs in water. The optimized method was validated and then applied to differentiate three HCHs from different manufacturers, to identify in situ degradation of HCHs of groundwater from a contaminated site and to resolve the carbon isotope fractionation occurring in the α-HCH oxidation by CaO₂/Fe(II) Fenton system. The results showed that the same reagents from different manufacturers have different carbon isotope compositions, and different isomers from the same manufacturer also have different isotope compositions, showing useful evidence in identifying the source of HCHs. The more enriched δ¹³C in the down-gradient wells indicated that HCHs have undergone biodegradation or/and chemical reactions in the groundwater system of the site. Carbon isotopic enrichment factors (ε_C) of −1.90 ± 0.10‰ were obtained in the oxidation process. Hence, the method validated in this study has great potential as a method for identifying the degradation of HCHs in a water environment. Full article

The study of the trophic relationships of aquatic animals requires correct estimates of their diets. We compared the quantitative fatty acid signature analysis (QFASA) and the isotope-mixing model IsoError, based on the compound-specific isotope analysis of fatty acids (CSIA-FA), which are potentially effective models for quantitative diet estimations. In a 21-day experiment, Daphnia was fed a mixture of two food items, Chlorella and Cryptomonas, which were supplied in nearly equal proportions. The percentages and isotope values of the FAs of the algal species and Daphnia were measured. The IsoError based on CSIA-FA gave an estimation of algae consumption using only one FA, 18:3n-3. According to this model, the proportion of consumption of Chlorella decreased while the proportion of consumption of Cryptomonas increased during the experiment. The QFASA model was used for two FA subsets—the extended-dietary subset, which included sixteen FAs, and the dietary one, which included nine FAs. According to both subsets, the portion of consumed Chlorella decreased from Day 5 to 10 and then increased at Day 21. The comparison of the two model approaches showed that the QFASA model is a more reliable method to determine the contribution of different food sources to the diet of zooplankton than the CSIA-based mixing model. Full article

Unintended pesticide pollution in soil, crops, and adjacent environments has caused several issues for both pesticide users and consumers. For users, pesticides utilized should provide higher yield and lower persistence while considering both the environment and agricultural products. Most people are concerned that agricultural products expose humans to pesticides accumulating in vegetation. Thus, many countries have guidelines for assessing and managing pesticide pollution, for farming in diverse environments, as all life forms in soil are untargeted to these pesticides. The stable isotope approach has been a useful technique to find the source of organic matter in studies relating to aquatic ecology and environmental sciences since the 1980s. In this study, we discuss commonly used analytical methods using liquid and gas chromatography coupled with isotopic ratio mass spectrometry, as well as the advanced compound-specific isotope analysis (CSIA). CSIA applications are discussed for tracing organic pollutants and understanding chemical reactions (mechanisms) in natural environments. It shows great applicability for the issues on unintended pesticide pollution in several environments with the progress history of isotope application in agricultural and environmental studies. We also suggest future study directions based on the forensic applications of stable isotope analysis to trace pesticides in the environment and crops. Full article

An innovative approach based on the combination of analytical pyrolysis coupled with gas chromatography-mass spectrometry (Py-GC/MS) with compound-specific isotope analysis (Py-CSIA) is used to study the composting process of maize biomass. This multidisciplinary approach aims to elucidate the decomposition rate of the main biogenic materials (lignin, cellulose, proteins, lipids, and waxes) responses to the composting process. According to Py-GC/MS data/structural composition, a noticeable and significant decrease during the first stage of the composting process of carbohydrates and aromatic compounds is found, followed by a gradual increase of all compounds till the end of the experiment. This trend, along with an increase of fatty acids methyl-ester at the first composting stage, sustains the microbial activity and its stabilization over time. Py-CSIA data showed a significant enrichment in ¹³C in all identified compounds over time, supporting the semi-quantitative results and the decomposition of initial biomass throughout the composting process. This trend is also perceptible in lignin moieties, long-chain aliphatic structures, and isoprenoids, as highly recalcitrant compounds, presumably due to depolymerization and carbon translocation of side-chain molecules during the composting process. Compound-specific isotope values showed a good correlation with the bulk isotope data, and this served as validation of the technique. However, bulk values showed higher heterogeneity because those represent an average of all organic compounds in the sample. By combining isotopic and structural information using Py-GC/MS and Py-CSIA, we are able to provide further information and a more detailed approach to the study of the decomposition process of biomass by considering the diverse dynamics of the main biogenic compounds. Full article

Natural attenuation (NA) processes represent a valuable option in groundwater remediation. At a heavily 1,2-dichloroethane (1,2-DCA) contaminated site, Compound-Specific Isotope Analysis (CSIA) in combination with Biological Molecular Tools (BMTs) were implemented as a rigorous characterization approach to evaluate the occurrence of Natural Attenuation in the proximity of the source area. By the use of microcosm experiments, the potential for natural and enhanced biodegradation under anaerobic conditions was documented, following the dichloroelimination pathway. Enrichment factors of −9.1‰ and −11.3‰ were obtained for ¹³C while Geobacter spp. and reductive dehalogenase genes (rdhs) were identified as main site-specific biomarkers. At pilot scale, enrichments of 13.5‰ and 6.3‰ for δ¹³C and δ³⁷Cl, respectively, high levels of reductive dehalogenase (rdh group VI) along with the dominance of Geobacter spp. indicated the occurrence of significant dichloroelimination processes in groundwater under anaerobic conditions. By using the site-specific enrichment factors, degradation extents over approximately 70–80% were estimated, highlighting the relevant potential of NA in 1,2-DCA degradation in the vicinity of the source area at the site. The proposed fine-tuned protocol, including CSIA and BMTs, is proven to be effective as a groundwater remediation strategy, properly assessing and monitoring NA at site scale. Full article