Stable Isotope Ratio Analysis Research Articles

Authentication of European sea bass according to production method and geographical origin by light stable isotope ratio and rare earth elements analyses combined with chemometrics.

In this work, stable isotope ratio (SIR) and rare earth elements (REEs) analyses, combined with multivariate data elaboration, were used to explore the possibility to authenticate European sea bass (Dicentrarchus labrax L.) according to: i) production method (wild or farmed specimens); ii) geographical origin (Western, Central or Eastern Mediterranean Sea). The dataset under investigation included a total of 144 wild and farmed specimens coming from 17 different European areas located in the Mediterranean Sea basin. Samples were subjected to SIR analysis (carbon and nitrogen) and REEs analysis (lanthanum, europium, holmium, erbium, lutetium, and terbium). Then, Analytical data were handled by Principal Component Analysis (PCA) and then by Orthogonal Partial Last Square Discriminant Analysis (OPLS-DA), to obtain functional classification models to qualitatively discriminate sea bass according to the conditions under study. OPLSDA models provided good correct classification rate both for production method and geographical origin. It was confirmed that chemometric elaboration of data obtained from SIR and REEs analyses can be a suitable tool for an accurate authentication of European sea bass.

Trophic transfer, biomagnification and risk assessments of four common heavy metals in the food web of Laizhou Bay, the Bohai Sea

This study investigated the trophic transfer of four common toxic metals (Cd, Cr, Cu and Hg) in the food web and assessed their potential ecological risks in Laizhou Bay, a spawning area for fishery populations in the Bohai Sea, North China. Based on the predation relations of 43 species that were representatives of the main trophic levels (TLs), a simplified food web was constructed using stomach content analysis and stable nitrogen isotope ratio (δ15N) analysis. Using copepods as the baseline species (TL = 2.00), the TLs of the organisms ranged from 1.96 (Polychaeta) to 4.47 (Japanese flounder) and showed the following ranking order: predatory pelagic or demersal fish > medium demersal fish > crustacean, cephalopod, small pelagic or demersal fish > zooplankton and Polychaeta. The metals showed different trophic transfer behaviors in the food web. Hg and Cr tended to be efficiently biomagnified between TLs, along the main food chains and in the food web. Cu biodiluted significantly with increasing TLs, while Cd showed no biomagnification or biodilution trends in the food web. At low or moderate levels of Cd and Hg, potential ecological risks were detected in the water and sediments at only a few sites, indicating their overall low ecological risks in the environment. The metals in the important fishery species (four top predatory fish, two cephalopods and eight crustaceans) were below the permissible limits, except for Cd in octopus and paddle crab, which reached or exceeded the most restrictive criteria. Based on the provisional tolerable weekly intake (PTWI) criteria, the safe weekly human consumption levels by humans of the predatory fish, cephalopods and crustaceans were species-specific (0.20–4.44 kg) and should be cautiously assessed.

Protura are unique: first evidence of specialized feeding on ectomycorrhizal fungi in soil invertebrates

BackgroundEctomycorrhizal fungi (ECM) play a central role in nutrient cycling in boreal and temperate forests, but their role in the soil food web remains little understood. One of the groups assumed to live as specialised mycorrhizal feeders are Protura, but experimental and field evidence is lacking. We used a combination of three methods to test if Protura are specialized mycorrhizal feeders and compared their trophic niche with other soil invertebrates. Using pulse labelling of young beech and ash seedlings we analysed the incorporation of 13C and 15N into Acerentomon gallicum. In addition, individuals of Protura from temperate forests were collected for the analysis of neutral lipid fatty acids and natural variations in stable isotope ratios.ResultsPulse labelling showed rapid incorporation of root-derived 13C, but no incorporation of root-derived 15N into A. gallicum. The transfer of 13C from lateral roots to ectomycorrhizal root tips was high, while it was low for 15N. Neutral lipid fatty acid (NLFA) analysis showed high amounts of bacterial marker (16:1ω7) and plant marker (16:0 and 18:1ω9) fatty acids but not of the fungal membrane lipid 18:2ω6,9 in A. gallicum. Natural variations in stable isotope ratios in Protura from a number of temperate forests were distinct from those of the great majority of other soil invertebrates, but remarkably similar to those of sporocarps of ECM fungi.ConclusionsUsing three in situ methods, stable isotope labelling, neutral lipid fatty acid analysis and natural variations of stable isotope ratios, we showed that Protura predominantly feed on mycorrhizal hyphae via sucking up hyphal cytoplasm. Predominant feeding on ectomycorrhizal mycelia by Protura is an exception; the limited consumption of ECM by other soil invertebrates may contribute to carbon sequestration in temperate and boreal forests.

C and H stable isotope ratio analysis using solid-phase microextraction and gas chromatography-isotope ratio mass spectrometry for vanillin authentication

It is possible to distinguish precious vanillin from Vanilla species (planifolia or tahitensis) from much less expensive synthetic and nature-identical vanillin on the basis of the stable isotope ratios of H and C (2H/1H, 13C/12C). Analysis is usually performed using GC-IRMS (Gas Chromatography - Isotope Ratio Mass Spectrometry) after solvent extraction of vanillin from the sample. Recently, head-space solid-phase microextraction (HS-SPME) has been proposed as an alternative for determining 13C/12C.The aim of this study was to develop a method to analyse 2H/1H in vanillin using SPME-GC-IRMS for the first time, by testing different operating conditions and comparing the results with those obtained after solvent extraction. The ultimate scope was to develop a quick, robust and effective method to measure 2H/1H and 13C/12C in vanillin to assess the authenticity of labelling.Almost 50 authentic samples from vanilla pods, nature-identical (ex) and synthetic vanillin and 4 commercial food products were taken into account. All the samples were subjected to HS-SPME-GC-IRMS analysis and most of them to GC-IRMS analysis after solvent extraction of vanillin.The SPME method developed for 2H/1H analysis guarantees the absence of isotopic fractionation, repeatability and reproducibility standard deviation of below 7‰ and savings in terms of time (from 30 to 5 min) and solvent.HS-SPME GC-IRMS analysis of δ2H and δ13C can be proposed as a rapid and robust method to discriminate different types of vanillin and assess the authenticity of natural vanillin, also contained in food matrices.

Isotopic and elemental profiles of Mediterranean buffalo milk and cheese and authentication of Mozzarella di Bufala Campana PDO: An initial exploratory study

Mozzarella di Bufala Campana (MBC) is a PDO cheese produced from whole buffalo milk in specific regions of southern Italy. Due to the high price and the limited amount of buffalo milk, MBC is potentially subject to mislabelling. Stable isotope ratio analysis combined with elemental analysis is one powerful technique for detecting the authenticity of PDO cheeses. Here, the elemental and isotopic profiles of authentic samples of buffalo milk and the corresponding MBC samples collected in the reference area in winter and summer are presented in an initial exploratory study. By merging MBC-PDO samples with non-PDO samples of buffalo mozzarella produced both inside and outside the reference area, a model was developed to classify product categories for this cheese. Despite the differences occurring during processing, along with differences in the season and production area, the model was effective in distinguishing PDO and non-PDO mozzarella, particularly when non-PDO cheeses were made outside the MBC reference area.

Supercritical Water Oxidation as a Pretreatment Method for Stable Carbon Isotope Ratio Analysis of Rice

Supercritical Water Oxidation as a Pretreatment Method for Stable Carbon Isotope Ratio Analysis of Rice

Compound-specific δ13C and δ15N analyses of fatty acids and amino acids for discrimination of organic, pesticide-free, and conventional rice (Oryza sativa L.)

Herein, we improve the procedure for organic rice authentication using compound-specific δ13C and δ15N analyses of fatty acids and amino acids, addressing the increasing demand for accurate methods to confirm organic authenticity. Organic rice (OR) and pesticide-free rice (PFR) featured higher values of δ15Nbulk than conventional rice (CR), whereas the corresponding differences between OR and PFR were insignificant. Additionally, OR, PFR, and CR could be discriminated based on some δ13Camino-acid and δ15Namino-acid values. δ13Cbulk was correlated with most δ13Cfatty-acid (r ≥ 0.596) values, and δ15Nbulk was strongly correlated with most δ15Namino-acid (r ≥ 0.834) values. The first component in the orthogonal projection to latent structure-discriminant analysis model allowed for a clear separation between OR and PFR, and good predictability (Q2Y = 0.506). Thus, the present study improves the reliability of organic authentication when bulk stable isotope ratio analysis alone is insufficient for the accurate discrimination of OR, PFR, and CR.

Dorset Pre-Inuit and Beothuk foodways in Newfoundland, ca. AD 500-1829.

Archaeological research on the Canadian island of Newfoundland increasingly demonstrates that the island’s subarctic climate and paucity of terrestrial food resources did not restrict past Pre-Inuit (Dorset) and Native American (Beothuk) hunter-gatherer populations to a single subsistence pattern. This study first sought to characterize hunter-gatherer diets over the past 1500 years; and second, to assess the impact of European colonization on Beothuk lifeways by comparing the bone chemistry of Beothuk skeletal remains before and after the intensification of European settlement in the early 18th century. We employed radiocarbon dating and stable carbon and nitrogen isotope ratio analysis of bulk bone collagen from both Dorset (n = 9) and Beothuk (n = 13) cultures, including a naturally mummified 17th century Beothuk individual. Carbon and nitrogen isotope analysis of 108 faunal samples from Dorset and Beothuk archaeological sites around the island were used as a dietary baseline for the humans. We combined our results with previously published isotope data and radiocarbon dates from Dorset (n = 12) and Beothuk (n = 18) individuals and conducted a palaeodietary analysis using Bayesian modelling, cluster analysis and comparative statistical tests. Dorset diets featured more marine protein than those of the Beothuk, and the diets of Beothuk after the 18th century featured less high trophic level marine protein than those of individuals predating the 18th century. Despite inhabiting the same island, Dorset and Beothuk cultures employed markedly different dietary strategies, consistent with interpretations of other archaeological data. Significantly, European colonization had a profound effect on Beothuk lifeways, as in response to the increasing European presence on the coast, the Beothuk relied more extensively on the limited resources of the island’s boreal forests and rivers.

Fatty Acid- and Amino Acid-Specific Isotope Analysis for Accurate Authentication and Traceability in Organic Milk.

The present study describes compound-specific δ13C and δ15N analyses of fatty acids and amino acids for improving the accurate authentication of organic milk (OM) against conventional milk (CM) collected in Korea. Most δ13Cfatty-acid and δ13Camino-acid values were lower in OM than in CM ( P < 0.05); however, most δ15Namino-acid values displayed weak discriminative power for OM authentication. Higher isotopic fractionation was observed in δ13Cfatty-acid than in δ13Camino-acid and δ15Namino-acid, with fractionation trends differing with individual amino acids. In particular, δ13Clinoleic-acid of -33.5‰ and δ13Cmyristic-acid of -28‰ were determined to be promising year-round threshold values for Korean OM authentication. The δ13Cbulk was highly correlated with δ13CAla ( r = 0.92) and δ13Coleic-acid, trans ( r = 0.77), and strong positive correlations were observed between δ13CVal and δ13CIle ( r = 0.98) and between δ15NThr and δ15NSer ( r = 0.90). Chemometric modeling for OM authentication produced a high quality model ( R2 X = 0.547, R2 Y = 0.865, and Q2 = 0.689) with reliable chemical markers, notably δ13Cmyristic-acid, δ13Clinoleic-acid, and δ13Cstearic-acid. Furthermore, the models developed for seasonal separation in OM ( Q2 = 0.954) and CM ( Q2 = 0.791) were of good quality. Our findings, based on compound-specific isotope data, improve the reliability of OM authentication in cases where bulk stable isotope ratio analysis alone is insufficient. They also provide valuable insight into the control of fraudulent OM labeling in Korea, with potential application in other countries.

Evidence of secondary sulfate production in the mineral soil of a temperate forested catchment in southern Québec, Canada

The analysis of sulfate stable isotope ratios (δ18O-SO4 and δ34S-SO4) in different hydrological compartments of forested catchments has revealed the major role of the humus layer in recycling atmospherically derived sulfur (S). The contribution of the mineral soil to S recycling is still uncertain and may vary among forest types. Here, seasonal variations in SO4 concentration, δ18O-SO4 and δ34S-SO4 were investigated over a period of two and a half years in precipitation and at various depths in the soil solutions in a temperate forest catchment dominated by sugar maple in southern Québec, Canada. δ18O-SO4 declined from precipitation (11.8‰) to the humus solution (4.8‰) and to the soil solution beneath the upper B-horizon (1.2‰). No decline was observed below the upper B-horizon. This decline from precipitation to the humus layer reflected a production of secondary sulfate through chemical oxidation of SO2 in the canopy, while the decline in the soil resulted from microbial mineralization of organic S. In contrast with findings at other boreal and temperate forest sites, lower δ18O-SO4 in the upper mineral soil than in the humus layer was indicative of microbial transformations of S not only in the humus layer but also deeper in the upper part of the mineral soil. Significant seasonal variations were found for δ18O-SO4 in precipitation and in soil solutions beneath the humus and the top mineral horizon, reflecting the influence of both hydrological and microbiological factors. Higher δ18O-SO4 in the soil solution in fall than in spring and summer resulted from the release of primary sulfate from snow cover in early spring, which was subsequently recycled by soil microorganisms during the growth season, resulting in a higher proportion of secondary sulfate in fall than in spring and summer.

Stable isotope ratio analysis (C, N, S) of hair from modern humans in Ethiopia shows clear differences related to subsistence regimes

We measured the stable isotope ratios (C, N, S) of modern human hair collected from 49 people in rural southern Ethiopia to characterize and compare the stable isotope signatures of interacting and ethnically distinct populations with differing economic practices (farmers, pastoralists, fishers) in order to determine if the dietary differences are visible and measurable in their hair isotope values. We found that there were significant differences in all three elements that can distinguish these economic practices (carbon: Χ2 = 8.523, p = 0.014; nitrogen: Χ2 = 35.372, p < 0.001; and sulfur: Χ2 = 30.887, p < 0.001). These results demonstrate the utility of isotopic methods as an indicator of dietary difference in living populations and shows that diverse dietary adaptations and economies even among interacting and neighboring peoples can be distinguished in this region of Ethiopia.

Multi-isotope approach of Pb, Cu and Zn in urban aerosols and anthropogenic sources improves tracing of the atmospheric pollutant sources in megacities

Studies including multiple isotope systems in aerosols promises unparalleled insights into sources and pathways of metals in the atmosphere. However, such studies remain rare because of the challenges associated with small sample sizes and low analyte masses of the target elements. Here, we present the first study combining accurate and precise determination of Pb, Cu and Zn isotopic ratios in aerosols and anthropogenic materials collected in São Paulo, Brazil. We use a sequential ion chromatography procedure with two different resins for the separation and purification of the analytes. Multi collector mass spectrometry is used for the accurate and precise determination of the isotope ratios. Long term analytical reproducibilities are ±0.035 for 206Pb/204Pb, ± 0.13‰ for δ65CuNIST and ±0.1‰ for δ66ZnJMC (±2σ). Accuracy is assessed using certified reference materials (CRM NIST 2783 aerossol, BRP-1 and others). We analyzed 57 source samples (road dust, tires, cement, road tunnel aerosol) and 113 aerosol samples collected between 2013 and 2015. The results for São Paulo are critically compared with previously published data from studies conducted in São Paulo, London and Barcelona. The key findings are: 1. The isotope signatures for Zn in tires (δ66ZnJMC = 0.16 ± 0.14, 2σ, n = 9) and road dust (δ66ZnJMC = 0.17 ± 0.19, 2σ, n = 13) are similar in São Paulo and London suggesting that this isotope system can be used as element specific tracers for non-exhaust traffic. 2. 206Pb/207Pb vs δ66ZnJMC and δ66ZnJMC vs δ65CuNIST multi-isotopic diagrams successfully separate wear off from cars including tires and brakes, car exhaust, industrial emissions and cement sources and improves the discrimination of air pollutant sources. 3. The source identification based on isotope ratios agrees source apportionment based on emissions inventory from these cities. 4. We present Pb, Cu and Zn isotopic data for the first time for the CRM NIST 2783 and BRP-1. These new data will enable future intercalibration and quality controls in other laboratories. Our study confirms that stable isotope ratio analysis have a great potential for element specific source characterization (e.g., separating non combustion traffic sources from combustion sources) for Cu, Zn and Pb.

Water movement via adventitious roots of the prostrate shrub Juniperus sabina in semiarid areas of China

The prostrate evergreen shrub Juniperus sabina L. is an important tree species for revegetation in semi-arid areas of China. The root system of J. sabina comprises a main root system, which can reach deeper soil, and adventitious roots that mainly extend in the surface soil. In our previous studies, water movement between different soil layers via these root systems associated with rainfall-induced vertical heterogeneity in soil moisture has been suggested by sap flow analysis. In this study, to confirm the existence of water movement from the deeper soil to the surface soil (i.e., hydraulic redistribution) under the progression of drying after rainfall, we controlled soil moisture in six potted J. sabina plants. On the assumption of drying after rain, the main roots continued to receive irrigation and only the irrigation on the adventitious root part was stopped. When drying had sufficiently progressed and the decline in soil moisture had stabilized, 18O-labeled water was irrigated at the main roots for 7-9 days. The 18O-labeled water was detected from the adventitious roots and soil on the adventitious root through the analysis of the oxygen stable isotope ratio of water. Sap flow was also measured during the experiment. Two sensors were implanted on each studied plant: one in the base of the decumbent stem, located between the main root system and adventitious roots, and the other distal to the decumbent stem. Sap flow occurred at nighttime only in the base of the decumbent stem during drying of the adventitious root. These results suggest the existence of water movement from moist soil to dry soil via the main and adventitious roots (hydraulic redistribution), which increased with the progression of drought.

Characterisation and attempted differentiation of European and extra-European olive oils using stable isotope ratio analysis.

European law requires a designation of origin for virgin and extra virgin olive oils (at least in terms of EU/non-EU provenance). Stable isotope ratios have been successfully applied to determine the geographical origin of olive oils, but never to distinguish EU and non-EU oils. In this study 2H/1H, 13C/12C and 18O/16O ratios were analysed in bulk olive oils using Isotope Ratio Mass Spectrometry (IRMS) as well as 13C/12C and 2H/1H in the four main fatty acids (linoleic, oleic, palmitic and stearic acids) using IRMS coupled with GC. The isotopic composition of olive oils was successfully used to distinguish samples originating in the two areas. Specifically, when bulk data were combined with fatty acid isotopic data the differentiation power of the method improved clearly. This separation is due to the specific isotopic fingerprint of the individual countries making up the EU and non-EU samples.

Combining microvolume isotope analysis and numerical simulation to reproduce fish migration history

Abstract Tracking the movement of migratory fish is of great importance for efficient conservation, although this has been technically difficult to achieve in small fish to which artificial tags cannot be attached. We show that migration history can be reproduced by combining high‐resolution otolith stable oxygen isotope ratio (δ18O) analysis and numerical simulation. High‐precision micromilling and microvolume carbonate analysing systems had the remarkable capability of extracting the otolith δ18O profiles with 10–30 days resolution. Furthermore, reasonable movements were reproduced by searching the routes consistent with the otolith δ18O profile, using an individual‐based model with random swimming behaviour. This method will be a valuable alternative to tagging and electronic loggers for revealing migration routes in early life stages, thereby providing crucial information to understand population structures and the environmental cause of recruitment variabilities, and to validate and improve fish movement models.

Custom-made PTFE filters for ultra-clean size-fractionated aerosol sampling for trace metals

Atmospheric deposition of marine aerosols is one of the important sources of dissolved trace metals in the ocean. Analyses of trace metals and stable isotope ratios in size-fractionated aerosol particles using extraction experiments are useful to understand the dissolution behavior and biogeochemical cycles of trace metals in the marine environment. These analyses require sample treatments in clean environment without contamination of target metals from the sampling filter. However, the contamination of target metals from sampling filter is not low enough to determine the fractional solubility (extractive metal/total metal) and the stable isotope ratio of trace metals. In this study, we optimized the acid-washing procedure of hydrophobic PTFE filter for the Sierra-type cascade impactor. The results showed that the filter blanks of the custom-made PTFE filters used for the upper stages (= PTFE-ST) in the cascade impactor were about 10–1000 times lower than those of cellulose filters. In addition, the filter blanks of major ions and water-soluble organic carbon were also decreased by using PTFE-ST. Trace metal concentrations in size-fractionated (7 fractions) aerosol particles with very short sampling periods have been successfully determined except for the backup stage. Thus, aerosol sampling with PTFE filter with only a tiny contamination is a useful method to obtain chemical and isotopic compositions of various elements in marine aerosol particles.

Independent trophic behavior and breeding success of a resident flycatcher and a coexisting migratory congener

AbstractHistorical biases towards the study of Palearctic and Nearctic bird migration systems has resulted in a dearth of information on the ecology of intratropical migrants, which likely also play important ecological roles within their communities. For instance, there is little information on the foraging ecology and breeding biology of co‐occurring migrants and residents within the intratropical migratory system. Thus, we used two congeneric flycatchers – the resident Plain‐crested Elaenia Elaenia cristata (Aves: Tyrannidae), and the migrant Lesser Elaenia E. chiriquensis (Tyrannidae) – to check whether either of them showed foraging niche plasticity mediating their coexistence and the effects of such syntopy on their breeding behavior and fitness. To do so, we used an ensemble of methods that included the evaluation of food resource phenology, foraging behavior observations, stable isotope ratio analysis in birds’ tissues, and nest monitoring. Our results confirmed that residents have foraging behavior plasticity but reveals its trophic behavior and breeding success is independent of the coexistence with migrants. Since such results depart from the predictions based on the current (and often Northern‐biased) theories of bird migration, we discuss alternative hypotheses explaining our findings, including the influence of physiology on diet and the behavioral responses of nest predators. Once there is a clear knowledge gap about the interspecific interactions between residents and intratropical migrants, our study represents a simple, yet important, step towards understanding the mechanisms underlying this system.

Online wet oxidation/isotope ratio mass spectrometry method for determination of stable carbon isotope ratios of water-soluble organic carbon in particulate matter.

Water-soluble organic carbon (WSOC) is formed by oxidation of organic compounds in particulate matter (PM) and accounts for 25-80% of the organic carbon in PM. Stable carbon isotope ratio (δ13 C) analysis is widely used to identify the sources of PM, but determining the δ13 C values of WSOC is complicated and requires a time-consuming pretreatment process. We have developed an online wet oxidation/isotope ratio mass spectrometry method with a reduced pretreatment time. We have measured the δ13 C values of WSOC by using this method. The method showed high accuracy (0.1‰) and precision (0.1‰) for levoglucosan, and the limit of detection was sufficiently low for WSOC analysis. Using this method, we determined δ13 C values of WSOC in PM2.5 samples collected in Japan during the period from July to November 2017 and found that the values ranged from -26.5‰ to -25.0‰ (average, -25.8‰). Our simple, low-blank method could be used for rapid quantitative analysis of the δ13 C values of WSOC in PM2.5 . We propose that this online method be used as a standard method for δ13 C analysis of WSOC.

The use of stable isotope ratio analysis to characterise saw palmetto (Serenoa Repens) extract

Saw palmetto extract (SPE) has many pharmacological effects. Thus, its demand and value has increased steadily, along with the presence of counterfeit SPEs on the market.In this work bulk δ13C, δ2H, δ18O and fatty acid δ13C, δ2H analysis was performed in 20 authentic and 9 commercial SPEs, 12 meat fats and 4 pure fatty acids.Authentic SPEs are characterised by bulk values from −31.0‰ to −29.7‰ for δ13C, −176‰ to −165‰ for δ2H, 27.2‰ to 40.7‰ for δ18O, and values of capric, caprylic, lauric, myristic, palmitic and oleic acids from −37.4‰ to −30.5‰ for δ13C and −187‰ to −136‰ for δ2H. The isotopic values of all the commercial SPEs were out of these ranges and more similar to those of meat fat and pure fatty acids.Stable isotope ratio analysis can therefore be proposed as a suitable tool for detecting adulteration in SPEs.

Combination of sugar and stable isotopes analyses to detect the use of nongrape sugars in balsamic vinegar must.

"Aceto Balsamico di Modena" (ABM) is a protected geographical indication salad dressing obtained from cooked and/or concentrated grape must, with the addition of wine vinegar and a maximum of 2% caramel (EU Reg 583/2009). In this study, we investigated whether the combination of 13 C/12 C of ethanol using isotope ratio mass spectrometry with 2 H-site-specific natural isotope fractionation-nuclear magnetic resonance and minor sugars using ion chromatography with pulse amperometric and charged aerosol detection is able to improve detection of sugar addition to ABM must. A large selection of authentic Italian grape musts and different samples of balsamic vinegar with an increasing percentage of added beet, cane, and sugar syrups were considered. The possible degradation of sugars in the ABM matrix during shelf life was also investigated. While stable isotope ratios analysis remains the favoured method for determining cane and beet sugar addition, dosage of minor sugar (in particular maltose) proved to be very useful for detecting the addition of sugar syrup. Thanks to this innovative approach, 3 out of 27 commercial ABMs were identified as adulterated. A combination of stable isotope ratio and ion chromatography with pulse amperometric and charged aerosol detection analysis can be therefore proposed as a suitable tool for detecting the authenticity of ABM must.