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Molecules
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Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives
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Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives

A Special Issue of Molecules (ISSN 1420-3049) belonging to the section "Analytical Chemistry".

Deadline for manuscript submissions: 31 December 2026 | Viewed by 4483

Editor

Dr. Matteo Perini

E-Mail Website
Guest Editor
Fondazione Edmund Mach, Via E. Mach 1, 38098 San Michele all’Adige, Italy
Interests: stable isotope analysis; traceability; authentication; mass spectrometry; method development

Special Issue Information

Dear Colleagues,

The stable isotopic composition of matter is governed by a wide range of physical, chemical, and biological mechanisms. Tracking the different transformations of such matter has become possible through the use of stable isotope analysis. This powerful tool provides insights into processes occurring at various spatial and temporal scales. Over the last decades, the development of increasingly accurate and sophisticated isotopic techniques has enabled the analysis of an exceptionally broad spectrum of matrices, including environmental (e.g., sediments, soils, waters, petroleum, meteorites), biological (e.g., bones, blood, organic tissues), and agri-food samples (e.g., milk, oils, wine). As a result, the application of stable isotope methodologies has rapidly expanded across numerous scientific disciplines, including geochemistry, archaeology, forensic science, biomedicine, environmental science, microbiology, soil science, and doping and fraud control, among many others. Indeed, the versatility and robustness of isotopic approaches make them a unique tool, with virtually limitless potential for unveiling new knowledge across the natural and applied sciences.

This Special Issue is dedicated to providing an updated and comprehensive view of recent advances in the application of stable isotope analysis within the diverse fields of chemistry. Contributions may include, but are not limited to, improvements in analytical procedures and instrumentation, such as new sample preparation strategies and innovative instrumental setups, as well as novel and emerging applications of isotope-based methods. Furthermore, studies that showcase recent applications of already well-established methodologies, yielding significant and novel results, are highly encouraged, along with review articles that synthesize current knowledge and provide future perspectives.

We are confident that this collection will be of great interest to the scientific community, reflecting the growing relevance and expanding role of isotopic analysis as a cornerstone in modern chemical and interdisciplinary research.

Dr. Matteo Perini
Guest Editor

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-anonymized peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Molecules is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • stable isotope analysis
  • compound specific stable isotopes
  • authenticity
  • traceability
  • biogenic isotopic composition
  • biogeochemistry
  • geochemistry
  • molecular archaeometry
  • environmental chemistry

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Published Papers (5 papers)

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Research

22 pages, 6769 KB  
Article
Origin and Function of Bitumen-Coated Torpedo Jars from the Sasanian to Early Islamic Period Fort of Fulayj in Oman
by Jacques Connan, Seth Priestman, Michael H. Engel, Alex Zumberge, Gazmend Elezi and Nasser S. Al-Jahwari
Molecules 2026, 31(15), 2571; https://doi.org/10.3390/molecules31152571 - 23 Jul 2026
Viewed by 520
Abstract
Geochemical and isotopic analysis of bitumen-lined transport container vessels, so called ‘torpedo jars’, from the Sasanian to Early Islamic period fort of Fulayj in Oman confirms the presence of two distinct compositional categories that can be matched to contemporary sources in different areas [...] Read more.
Geochemical and isotopic analysis of bitumen-lined transport container vessels, so called ‘torpedo jars’, from the Sasanian to Early Islamic period fort of Fulayj in Oman confirms the presence of two distinct compositional categories that can be matched to contemporary sources in different areas of southwest Iran. It appears that the bitumen used to line jars was extracted from different geographic areas, hinting at the existence of multiple production locations for this vessel class. In terms of the function of the jars, organic residue analysis provides a positive identification of biomarkers associated with the storage and transportation of red wine. These results provide the first published confirmation of the long-suspected association between torpedo jars and wine transportation from the core urbanised zone of the Sasanian and Early Islamic imperial heartlands towards the Persian Gulf and Indian Ocean maritime sphere. The results have added significance as the samples are derived from relatively accurately dated archaeological contexts spanning the period before and after the Islamic conquest. Full article
(This article belongs to the Special Issue Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives)
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27 pages, 1810 KB  
Article
A Multi-Isotope Approach (δ2H, δ18O, δ13C, δ15N) for Discriminating Raspberry Production Systems and Assessing Agroecosystem Functioning
by Roxana Elena Ionete, Diana Costinel, Ana Maria Simionescu, Marius Gheorghe Miricioiu, Augustina Pruteanu, Aura Irina Istrate and Oana Romina Botoran
Molecules 2026, 31(14), 2459; https://doi.org/10.3390/molecules31142459 - 14 Jul 2026
Viewed by 500
Abstract
The development of sustainable and climate-resilient food systems increasingly relies on robust analytical methodologies capable of integrating environmental, biochemical, and management-related signals. In this study, a multi-isotope framework based on δ2H, δ18O, δ13C, and δ15N [...] Read more.
The development of sustainable and climate-resilient food systems increasingly relies on robust analytical methodologies capable of integrating environmental, biochemical, and management-related signals. In this study, a multi-isotope framework based on δ2H, δ18O, δ13C, and δ15N was applied to assess its capacity to discriminate between contrasting raspberry production systems and to provide chemically grounded indicators of agroecosystem functioning. Raspberry fruits (Rubus idaeus L.; cultivars Opal and Delniwa) were collected during the 2024–2025 growing seasons from two distinct systems in Romania: an organic open-field system and a rainfed agroforestry system. Stable isotope ratio analysis revealed system-dependent isotopic patterns, with the strongest differentiation observed for δ15N. Nitrogen isotope composition (δ15N) provided the strongest discrimination, with enriched values in organic fruits (2.73–9.77‰) and depleted values in agroforestry fruits (−3.01 to 0.62‰), reflecting differences in nitrogen sources and cycling pathways. Hydrogen and oxygen isotopes (δ2H: −60.46 to −4.62‰; δ18O: −6.19 to 10.41‰) were consistent with hydroclimatic variability and evaporative fractionation processes associated with soil–plant–atmosphere interactions. Carbon isotopes (δ13C: −28.14 to −22.62‰) provided complementary insights into plant water-use conditions. Multivariate statistical analysis supported the separation between production systems, while short-term fertilisation effects were secondary to system-level controls. The results suggest that raspberry fruits preserve an integrated isotopic fingerprint of production environment and management practices. From an analytical chemistry perspective, this work highlights the relevance of multi-isotope approaches as transferable tools for food authentication, traceability, and sustainability assessment, contributing to the broader application of stable isotope techniques across complex biological systems. Full article
(This article belongs to the Special Issue Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives)
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14 pages, 1909 KB  
Article
Determining the Authenticity of Ghanaian Honeys Using Stable Isotope Ratio Analysis (SIRA)
by Lebene Kpattah, Zala Sel, Marjeta Mencin, Dennis Kpakpo Adotey and Nives Ogrinc
Molecules 2026, 31(14), 2401; https://doi.org/10.3390/molecules31142401 - 8 Jul 2026
Viewed by 569
Abstract
Honey is a high-value food product that is vulnerable to adulteration with exogenous sugars, posing challenges for food authenticity and consumer protection. This study applied Stable Isotope Ratio Analysis (SIRA) to assess the authenticity of honey collected from three major honey-producing regions of [...] Read more.
Honey is a high-value food product that is vulnerable to adulteration with exogenous sugars, posing challenges for food authenticity and consumer protection. This study applied Stable Isotope Ratio Analysis (SIRA) to assess the authenticity of honey collected from three major honey-producing regions of Ghana (Volta, Bono and Bono East). A total of 28 honey samples were analysed by elemental analysis–isotope ratio mass spectrometry (EA-IRMS) to obtain carbon (δ13C), nitrogen (δ15N) and sulphur (δ34S) isotope composition. Honey authenticity was evaluated according to AOAC Official Method 998.12 by comparing δ13C values of bulk honey and the corresponding protein fraction. The δ15N and δ34S values in honey protein were used to investigate environmental and regional variability. Samples without detectable C4 adulteration exhibited δ13Cprotein values consistent with C3 floral sources, whereas several samples showed Δδ13C values more negative than −1.0‰, indicating the presence of C4-derived sugars above the AOAC adulteration threshold. Calculated C4 sugar contents ranged from 8 to 12% in moderately adulterated samples to as high as 78–79% in severely adulterated samples, confirming substantial dilution with C4 sugars. Nitrogen and sulphur isotope ratios provide additional information on environmental and regional variability among the sampled regions. Principal Component Analysis revealed that the first two principal components (PC1 and PC2) accounted for 83.8% of the total variance 83.8% of the total variance and showed separation between samples with detectable C4 adulteration and those without, while highlighting regional isotopic differences. These results demonstrate that stable isotope analysis is an effective tool for detecting C4 sugar adulteration in honey and that the combined use of carbon, nitrogen and sulphur isotopes can provide additional information on environmental and regional variability. These findings provide preliminary isotopic data on honey collected from three major honey-producing regions of Ghana and support the application of the stable isotope approach for honey authenticity assessment and quality control. Full article
(This article belongs to the Special Issue Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives)
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15 pages, 5026 KB  
Article
Isoscape of Oxygen Stable Isotopes in Woods of the Amazon
by Ana Claudia Gama Batista, Maria Gabriella da Silva Araújo, Isabela Maria Souza-Silva, Deoclécio Jardim Amorim, Fabiana Cristina Fracassi Adorno, Gabriela Bielefeld Nardoto, Vladimir Eliodoro Costa, Mario Tomazello-Filho, Niro Higuchi, Perseu da Silva Aparicio, Yasmin Lara Bezerra Vieira da Silva, Marta Silvana Volpato Sccoti, Ana Carolina Barbosa, Fabio José Viana Costa, João Paulo Sena-Souza, Gabriel J. Bowen and Luiz Antonio Martinelli
Molecules 2026, 31(9), 1542; https://doi.org/10.3390/molecules31091542 - 6 May 2026
Viewed by 1160
Abstract
Stable oxygen isotopes (δ18O) in wood provide integrative records of plant water use and regional hydroclimatic processes, offering a powerful framework for spatial ecological analysis in tropical forests. Here, we present the first regional-scale δ18O isoscapes for Amazonian [...] Read more.
Stable oxygen isotopes (δ18O) in wood provide integrative records of plant water use and regional hydroclimatic processes, offering a powerful framework for spatial ecological analysis in tropical forests. Here, we present the first regional-scale δ18O isoscapes for Amazonian wood based on 387 trees sampled across 25 sites. After α-cellulose extraction, δ18O values were modeled using multiple linear regression (MLR) and Random Forest (RF) approaches. A Moran’s I test revealed no significant spatial autocorrelation (p = 0.73), indicating that geostatistical interpolation methods such as kriging were not appropriate for this dataset. The MLR model based on site-average data achieved an R2 of 0.70, with a mean absolute error (MAE) of 0.56‰ and root mean square error (RMSE) of 0.68‰. The RF model showed comparable performance (R2 = 0.67; MAE = 0.64‰; RMSE = 0.77‰). Both approaches reproduced a coherent southeast-to-northwest gradient, with lower δ18O values in the western Amazon and higher values in the east, consistent with regional patterns in precipitation isotopic composition and evapotranspiration. These findings demonstrate that climate-driven statistical modeling effectively captures large-scale isotopic structure across the Amazon basin, providing a robust spatial representation of δ18O variability in tropical forest wood. Full article
(This article belongs to the Special Issue Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives)
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13 pages, 435 KB  
Article
Influence of Diet and Growth Conditions on the Carbon and Nitrogen Stable Isotopic Composition of Aspergillus niger Mycelium: Insights for Fungal Chitosan Characterization
by Matteo Perini, Raffaele Guzzon, Silvia Pianezze, Francesca Violardo and Roberto Larcher
Molecules 2025, 30(20), 4142; https://doi.org/10.3390/molecules30204142 - 21 Oct 2025
Viewed by 1000
Abstract
This study investigates, for the first time, the relationship between carbon (δ13C) and nitrogen stable isotopic composition of Aspergillus niger mycelium, used as chitin and chitosan sources, and the fungus diet under controlled cultivation conditions. Four diets were tested, combining [...] Read more.
This study investigates, for the first time, the relationship between carbon (δ13C) and nitrogen stable isotopic composition of Aspergillus niger mycelium, used as chitin and chitosan sources, and the fungus diet under controlled cultivation conditions. Four diets were tested, combining different carbon (C3- and C4-glucose) and nitrogen sources (KNO3 and NH4Cl). Results showed that carbon sources significantly influenced δ13C values of the mycelium: C4-glucose diets led to more negative Δ13C values (δ13CMYCELIUM-δ13CDIET) compared to C3-glucose diets. Nitrogen sources also affected isotopic fractionation, with KNO3 leading to negative Δ15N (δ15NMYCELIUM-δ15NDIET) and NH4Cl yielding positive Δ15N. Conversely, pH and temperature showed negligible effects on δ15N, while continuous aeration during growth significantly decreased δ15N, possibly due to partial assimilation of atmospheric nitrogen. These findings demonstrate that both nutrient and cultivation parameters can modulate the isotopic fractionation in A. niger, particularly for nitrogen. Although a direct correlation between diet composition and δ15N could not be established, this work provides the first experimental link between fungal metabolism and its isotopic fingerprint. The results offer a scientific foundation for applying stable isotope ratio analysis to authenticate and trace fungal-derived chitin and chitosan, with potential applications in food and winemaking industries. Full article
(This article belongs to the Special Issue Stable Isotope Techniques Across Chemistry: Methods, Applications, and Perspectives)
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